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ISSN 2277 – 4289 www.gjrmi.com Editor-in-chief Dr Hari Venkatesh K Rajaraman

Managing Editor Dr. Shwetha Hari

Administrator & Associate Editor Miss. Shyamala Rupavahini

Advisory Board Prof. Rabinarayan Acharya Dr. Dinesh Katoch Dr. S.N.Murthy Dr. Mathew Dan Mr. Tanay Bose Dr. Nagaraja T. M. Prof. Sanjaya. K. S. Dr. Narappa Reddy

Editorial board Dr. Kumaraswamy Dr. Madhu .K.P Dr. Sushrutha .C.K Dr. Ashok B.K. Dr. Janardhana.V.Hebbar Dr. Vidhya Priya Dharshini. K. R. Mr. R. Giridharan

Honorary Members - Editorial Board Dr. Shubha Ganguly Dr Farhad Mirzaei Mr. Harshal Ashok Pawar


INDEX Medicinal Plant Research Clinical Research – Life sciences EFFECT OF NATURO FRUIT BARS ON BLOOD GLUCOSE LEVELS IN PATIENTS WITH TYPE-II DIABETES MELLITUS Nagaraju I, Reddy K Kodandarami, Giriprasad B, Lakshminarayanan G, Reddy C Damodar……………………………………………………………………………………….202–210 Biochemistry HYPOGLYCEMIC EFFECTS OF AQUEOUS AND ETHANOLIC EXTRACTS OF DANDELION (TARAXACUM OFFICINALE F.H. WIGG.) LEAVES AND ROOTS ON STREPTOZOTOCININDUCED ALBINO RATS Chinaka Nnamdi C, Uwakwe A A and Chuku L C…………………………………………...211–217 Pharmacognosy ANTIBACTERIAL EVALUATION OF ETHANOLIC EXTRACT OF CYNODON DACTYLON (L.) PERS. Kashyap Pranita, Sawarkar Hemant A, Mishra Koushlesh K………………………….218–224 Botany SACRED TREES OF TEMPLES OF TIRUCHIRAPPALLI, TAMIL NADU – THE NATURAL AND ECOLOGICAL HERITAGE OF INDIA Umavathi R, Parvathi A…………………………………..……………………………...…225–233 Life sciences CERTAIN INDIAN MEDICINAL PLANTS SAFEGUARD AGAINST CANCER Nema Rajeev, Khare Sarita, Jain Parul, Pradhan Alka…...…………………………..…..234–241 Botany ETHNO BOTANICAL SURVEY OF MEDICINAL PLANTS OF SARGODHA REGION AND ITS VICINITIES Ghani Abdul, Hasan Naqi, Ishtiaq M…………………………..........................................242–246

Indigenous Medicine Ayurveda EFFICACY OF GLYCYRRHIZA GLABRA LINN IN LEARNING, MEMORY AND COGNITIVE ACTIVITY – CURRENT FINDINGS AND FUTURE AVENUES Sheshagiri Srihari , Patel Kalpana Shanthibhai, Shrikrishna Rajagopala ………….…..247–257

COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – TENDER LEAVES OF STRYCHNOS NUX-VOMICA L., LOGANIACEAE PLACE – KOPPA, CHIKMAGALUR DISTRICT, KARNATAKA, INDIA


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Original Research Article EFFECT OF NATURO FRUIT BARS ON BLOOD GLUCOSE LEVELS IN PATIENTS WITH TYPE-II DIABETES MELLITUS Nagaraju I1, Reddy K Kodandarami2, Giriprasad B3, Lakshminarayanan G4, Reddy C Damodar5* 1,2,5

Division of Clinical Research, Sugen Life Sciences Pvt Ltd, Tirupati - 517 505, Andhra Pradesh, India. Srinivasa Diabetics and Medical Centre, Tirupati - 517 501, Andhra Pradesh, India. 4 Naturo Food and Fruit Products Pvt. Ltd, Ekarajapura village, Bangalore -562 114, India 3

*

Corresponding Author: E-mail: cdr@sugenlife.com; Ph/fax: 91-877-2276118 Received: 26/04/2012; Revised: 21/05/2012; Accepted: 28/05/2012

ABSTRACT The effect of Naturo Fruit Bars in altering metabolic parameters in type-2 diabetes was tested in a randomized open-label clinical trial on healthy volunteers and type-2 diabetics. To determine the safety of the fruit bars, 13 healthy volunteers and 10 type-2 diabetics were administered with 2, 3, 4 fruit bars on day 0, 7 and 14 and blood glucose levels were determined at time ‘zero’ and two hours after consumption. In the efficacy study 18 type-2 diabetics consumed two bars of apple flavor without added sugar per day for 12 weeks. Blood glucose, insulin, hemoglobin A1C, lipid profile and adiposity were determined on day zero and at the end of study. Results of the safety study indicated that consumption of 2, 3 or 4 fruit bars did not elevate blood glucose levels in both healthy and type2 diabetes subjects. Consumption of two bars per day for 12 weeks in type-2 diabetics did not increase the blood glucose levels. The levels of cholesterol, HDL cholesterol, LDL cholesterol, triglycerides and Body Mass Index did not alter. Consumption of Naturo fruit bars did not increase the blood glucose levels and thus can be used as a substitute for healthy food in type-2 diabetics without any undesirable effects.

Keywords: Naturo Fruit Bar, post-prandial blood glucose, lipid profile, insulin, HbA1C

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INTRODUCTION Diabetes mellitus is a metabolic disorder and its prevalence is an important global health problem (King et al., 1998). In 1985 an estimated 30 million people world-wide had diabetes, and in 2010, there were 285 million cases. This number is projected to increase to about 439 million by 2030 (Shaw et al., 2010). The prevalence of diabetes is higher in developed countries than in developing countries, but the incidence of diabetes in developing countries is likely to increase at a higher rate (Mohan V et al., 2010). Increased urbanization, economic development and job related sedentary habits in developing countries are likely the contributory factors for the expected rise in the cases of diabetes (Wasir JS and Misra A, 2004). In recent years, several nutritional supplements specifically designed for diabetic patients have been commercialized, which are unlikely to increase the post-prandial blood glucose levels compared to glucose or high glycemic foods (Fix et al., 2000). Liquid nutritional supplements (e.g. Glucerna) have been shown to reduce the post-prandial blood glucose levels in diabetes patients (Gonza´lezOrtiz M et al., 2000). Many snacks/bars are available as nutritional supplements for diabetic subjects, but their effect on blood glucose levels, insulin secretion and glycated hemoglobin (HbA1C) and lipid profile in diabetes has not been examined systematically. The aim of the present study was to investigate the effect of chronic administration of Naturo fruit bars (manufactured by Naturo Food and Fruit products Private Ltd, Bangalore, India) on metabolic parameters in patients with type-2 Diabetes mellitus. Earlier we determined the Glycemic Index (GI) value of Naturo fruit bar (apple flavor) and found it to be 38.50 [Reddy KK et al., 2009]. There are very limited supplements available in India to cater to the needs of the increasing number of diabetes

population, who are concerned and selective in taking appropriate snacks. It is anticipated that NFB are likely to serve as a healthy food choice for diabetic population in India. MATERIALS AND METHODS Investigational Product Naturo fruit bars (Photo slide 1) were prepared from fresh fruits by a unique process without any artificial additives and available in different flavors (Apple, Mango, Pineapple and Guava) containing with or without added sugars. The present study is focused on the Naturo Fruit bar, apple flavor without added sugars. Nutritional information of the Naturo fruit bar (apple flavor): Serving size: one fruit bar (20 g): calories (65.48 KCal), protein (0.35 g), carbohydrate (15.73 g), sugar (13.48 g), total fat (0.15 g), saturated fat (0.0004 g), dietary fiber (1.55 g), sodium (10 mg), potassium (120.92 mg), calcium (0.21 mg), Iron (0.91 mg), vitamin C (2.4 mg) and vitamin A (43.51 IU). In view of low carbohydrate, fat and sugar content, but enriched with fiber and protein, the NFB can be considered as a substitute for healthy food in type-2 diabetic patients. Study design A randomized open-label, clinical trial was carried out on normal healthy volunteers and type-2 diabetic patients at Sugen Life Sciences Pvt Ltd, Tirupati, India. The protocol for the study was adhered to Good Clinical Practice (GCP) guidelines and followed the recommendations of the World Medical Association Declaration of Helsinki. The study protocol was approved by an Independent Ethics Committee (IEC) of of Snehal Hospitals, Bombay, India. Informed Consent Form (ICF) was obtained from all healthy volunteers and diabetes patients and all subjects were closely monitored and recorded for any adverse events throughout the study.

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Photo Slide 1: Displaying the market available Naturo Fruit Bars

Subject selection parameters The clinical trial was initiated to assess the safety in healthy volunteers and diabetic patients followed by efficacy study in diabetic subjects only. In safety study, dose escalation and the safety of the NFB was determined in 13 adult healthy volunteers and 10 adult type type-2 diabetes subjects. In efficacy study, NFB was administered for 12 weeks in 18 type 2 diabetics to monitor the levels of blood glucose, insulin, HbA1C and lipid profile. There were no drop-outs outs in the present study. The age range of the subjects in the present study is 30–70 70 years. Both healthy volunteers and type-22 diabetes subjects were non non-obese (BMI < 25 kg/m2), non-smoking smoking and non nonalcoholic. All the healthy volunteers were in good health, as determined by medical history and physical sical examination; no history of hypertension, hepatic and renal disease, coronary artery disease, and or type type-1 or 2 Diabetes mellitus was reported. The inclusion criteria for diabetic subjects were those taking standard medication of Oral Hypoglycemic Agents gents (OHA) advised by the physician for the past three months, whose fasting blood glucose levels are maintained in the range of 100 100– 120 mg/dL and otherwise normal in other clinical and laboratory parameters. The exclusion criteria: Diabetic patients curr currently under treatment for heart disease, cancer, post postsurgery patients, tuberculosis and other chronic diseases, subjects who are bedridden for chronic diseases for prolonged time and kidney diseases, users of drugs which exert

intoxication and pregnant women. wo The duration of diabetes in the study subjects was between 3–7 7 years. During the study period the subjects were given clear instructions by the physician not to deviate from their regular drug intake, diet regime and physical exercise. Throughout the study, principal investigator and clinical research coordinator have monitored the subjects at regular intervals for any events. Weight and height were recorded with the subjects wearing light clothing without shoes. Height was measured and rounded off to the nearest centimeter,, with the subjects standing in erect posture. Body Mass Index (BMI) was calculated as weight in kg divided by the height in meter square. Waist girth was measured at the level of umbilicus with person breathing silently and hip measured asured as standing interinter trochanteric girth according to the method specified by Weiner and Lourie (Weiner JS and Lourie JA et al., 1981). Waist Hip Ratio (WHR) was calculated from the circumferences of waist and hip. Systolic and diastolic pressures were considered in Korotkoff phase I and V respectively as specified by Rose et al., (Rose GA et al., 1982). Diabetic subjects were requested to withdraw drug intake prior to blood sample collection. Fasting (11h) venous blood collected from the patients was allowed to clot for 30 min at room temperature and then centrifuged. The resulting plasma was separated into two aliquots. The first aliquot was used for the measurement of glucose, total cholesterol, high-density density lipoproteins (HDL), triglycerides and HbA1C. HbA1 Low-density lipoprotein cholesterol was calculated

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according to the Frieldwald et al., (1982) formula (total cholesterol- HDLC/5 + TG). The second aliquot was frozen at −200 C and processed within 30 days for the estimation of insulin by radioimmunoassay method with an intra- and inter-assay coefficient of variation of < 4.4 and 6.9 % respectively. Blood glucose was determined by the glucose-oxidase method with an intra- and inter-assay coefficient of variation of < 1 %. Serum lipid levels (total cholesterol, HDL cholesterol and triglycerides) and HbA1C were measured by enzymatic methods with an intra- and inter-assay coefficient of variation of < 3 %. The HDL cholesterol was estimated after selective precipitation of non-HDL fractions. In the safety study, all the subjects consumed an iso-caloric diet (69 Kcal, 15 g of carbohydrate, 1.2 g of protein, 0.2 g of total fat) through the personal substitution by the investigators on scheduled visit to the clinic (day 0, 7 and day 14). After the withdrawal of first blood sample, an iso-calorie diet was provided to all the subjects apart from Naturo fruit bars. Subjects were provided with 2, 3, 4 Naturo fruit bars on day 0, 7, 14 respectively (one time supplementation) and allowed to consume the bars in the clinic itself after taking the breakfast, under the supervision of the physician. To ascertain the safety of the NFB we conducted dose escalation study of fruit bars initially in the healthy volunteers followed by the type-II diabetes subjects. After the intake of fruit bars the subjects stayed in the clinic and were under the supervision of the physician and the blood was withdrawn after two hours of consumption of NFB. In addition these volunteers did not report any adverse effects after completion of the short-term study. Blood samples were drawn (fasting) and after two hours upon consumption of Naturo fruit bars for the estimation of glucose levels. In the efficacy study, 18 adult type-2 diabetes patients were recruited and instructed to consume two NFB / day for 12 weeks. The protocol adopted for efficacy study is same as in safety study. In the diabetes patients (safety study), postprandial blood sugar levels (Table-2) were reduced after the consumption of two Naturo

fruit bars. But the consumption of three and four bars moderately elevated the post-prandial blood sugar. This allowed us to select two fruit bars for efficacy study. Fasting and postprandial blood glucose levels were monitored weekly. On day ‘zero’ and on 12th week, fasting and post-prandial blood glucose, lipid profile (Cholesterol, HDL-Cholesterol, and Triglycerides), HbA1C and insulin levels were determined. Blood pressure, pulse rate, overall and abdominal obesity were recorded. Analysis Statistical analysis was performed using the statistical package (SPSS) version 16.0. Sample size was calculated using a clinical trial formula as described (Jayeseelan L and Rao PSS, 1989). Utilizing a standard deviation of 0.7, a sample size of 18 subjects was chosen to detect the effect with 80% power and a twosided significance level of 5 % was adopted. Initially the data was subjected to normality distribution. In descriptive statistics, results were expressed as mean ± SD unless otherwise specified. One-way analysis of variance (ANOVA) and student’s‘t’ test was applied where necessary. RESULTS The demographic and metabolic variables for the safety study are shown in table-1 for healthy volunteers and diabetes patients. As shown in table-1 no significant difference was observed in the overall obesity between healthy volunteers compared to type-2 diabetic patients, however, a significant difference was observed in abdominal obesity (p < 0.05). Mean systolic blood pressure was higher in diabetes patients (118.45 ± 6.82 vs 113.11 ± 10.28). Mean total cholesterol (163.70 ± 27.93 vs 181.03 ± 23.49) and LDL cholesterol (101.45 ± 28.67 vs 117.23 ± 20.92) were lower in diabetic patients when compared to healthy volunteers with no change in triglycerides and HDL cholesterol and there were no adverse events throughout the study. These results indicate that all the subjects, both healthy volunteers and diabetes patients, have similar overall metabolic profiles.

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Table.1 Demographic and metabolic profile of the study subjects (data as Mean ± SD) Characteristic value Age (years) Body Mass Index (Kg) Waist Hip Ratio (WHR) Systolic blood pressure Diastolic blood pressure Pulse rate Total cholesterol HDL cholesterol LDL cholesterol Triglycerides *p<0.05

Healthy volunteers (N=13) 37.28 ± 7.16 20.85 ± 2.30 0.848 ± 0.06 113.11 ± 10.28 78.03 ± 5.67 69.97 ± 4.91 181.03 ± 23.49 41.58 ± 5.68 117.23 ± 20.92 119.76 ± 38.93

Changes in the post-prandial blood glucose levels upon the consumption of NFB in both healthy and diabetic subjects were shown in table-2. These results show that consumption of two NFB decreased the post-prandial blood glucose (P < 0.0001) in diabetic subjects. Further, administration of 3 and 4 NFB bars moderately elevated the post-prandial glucose; however this elevation did not increase the base-line post-prandial glucose levels in diabetic subjects. In healthy volunteers the consumption of two NFB decreased the blood

Type-II Diabetics (N=10) 45.80 ± 5.27 20.92 ± 2.60 0.890 ± 0.04* 118.45 ± 6.82 78.95 ± 5.23 70.80 ± 4.66 163.70 ± 27.93 42.50 ± 4.77 101.45 ± 28.67 126.95 ± 44.26

glucose. Subsequent administration of 3 and 4 fruit bars showed an elevation from the base line glucose levels with some fluctuations in healthy volunteers. Overall, the F-values indicate that the fluctuation in post-prandial blood glucose levels from the base line values is significant in both healthy and diabetic subjects. Because two fruit bars were preferred by a majority of the subjects, this dose was chosen for the efficacy study (12-week) in type-2 diabetes patients.

Table.2 Mean values of fasting and post-prandial blood glucose levels in healthy and diabetic subjects (data as Mean ± S.D) Healthy Volunteers

Type II Diabetes

(N=13)

(N=10)

Fasting

74.0 ± 10.6

117.2 ± 8.5

Postprandial (2hrs) after test glucose:

96.3 ± 14.2*

145.1 ± 9.1*

2 NFBs

91.9 ± 13.0*

126.1 ± 9.0

3 NFBs

95.3 ± 11.3*

129.0 ± 7.4

4 NFBs

99.0 ± 14.3*

131.6 ± 6.4*

*F value for post-prandial blood glucose in healthy subjects=7.60; P < 0.05 *F value for post-prandial blood glucose in diabetic subjects=15.44; p < 0.05

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Mean differences for overall and abdominal obesity as well as on blood pressure did not show significant variation between day ‘zero’ and at the end of the 12--week study. Significant (p < 0.05) reduction in fasting blood glucose levels from day ‘zero’ to 12 weeks was observed in type-22 diabetes patients. There was a decrease in HbA1C (p < 0.11) and an increase in insulin levels between day ‘zero’ and towards the end of the study (p < 0.08). The extremities xtremities in mean difference (95 % CI) with altered S.E failed to show significant difference for HbA1C and insulin between day ‘zero’ and at the end of the study. Similarly, there was no significant changes in lipid profile (Cholesterol, HDL-cholesterol, LDL-cholesterol cholesterol and

triglycerides) in the subjects following the consumption of two fruit bars for 12 weeks. Alterations in blood glucose levels in typetype 2 diabetes subjects upon consumption of two NFB for twelve weeks are shown in Fig.1. A gradual decrease ase in both fasting and postpost prandial blood glucose levels was observed in diabetic patients upon consumption of NFB. Overall these results indicate that the window between the fasting and post-prandial post glucose levels decreased from day ‘zero’ to 12th week (p < 0.05). In order to attain the linear growth rate for both fasting blood sugar (FBS) and post-prandial (PPBS) S) a regression line is fitted and observed that the rate of decrease for FBS is – 0.7 and for PPBS S it is −2.0.

Figure 1: Alterations in blood glucose levels in type type-2 2 diabetes patients administered with Naturo Fruit Bars for twelve weeks

Fig 1

FBS

PPS

180.0

Blood glucose (mg)

160.0

140.0 y = -2.9328x + 168.78 120.0

100.0 y = -1.0157x + 116.14 80.0

60.0 1

2

3

4

5

6

7

8

9

10

11

12

Visits (in weeks)

Changes in the levels of fasting and post post-prandial blood glucose levels in type-2 2 diabetes patients administered with two Naturo fruit bars for a period of 12 weeks. Each point in the graph represents mean values observed on every week. One-way way analysis of variance was applied to find out the chan changes ges in fasting blood glucose (F(F value = 2.252: P < 0.01) and post-prandial prandial blood gluco glucose (F-value = 4.109: P < 0.001). The regression equation is  /  )*100} for FBS = − 0.7 and for PPBS specified for FBS and PPBS S separately. Linear Growth Rate {( PP = − 2.0

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DISCUSSION Since prevalence of type-2 diabetes mellitus is increasing, offering nutritional supplements that improve glycemic control without any adverse effects is critical in the long- term management of diabetes patients. In the present study, the safety of NFB consumption in type-2 diabetes patients was evaluated in a well-controlled clinical trial. No adverse events were recorded throughout the study. The results presented in the safety study demonstrate that, consumption of two/three/four NFB did not increase the postprandial blood glucose levels in healthy volunteers and diabetes subjects. Based on the results of the safety study, an efficacy study over a period of 12 weeks (two fruit bars/day) was performed. The objective of this study was to determine whether consumption of NFB for longer periods of time has any effect on blood glucose levels and metabolic profile in type-2 diabetes patients. In addition, this study allowed us to monitor for any adverse effects and changes in key biochemical parameters. The results of the long-term study demonstrate that consumption of two NFB everyday over a period of 12 weeks did not increase the blood glucose levels and has no adverse effects in type-2 Diabetes mellitus patients. Interestingly, NFB produced a significant decrease in both fasting and postprandial blood glucose levels after 12 weeks of consumption compared to day ‘zero’ (Fig.1). These results are in agreement with earlier studies by Gonza´lez-Ortiz et al., 2006, that nutritional supplements containing carbohydrates of low glycemic value has decreased the post-prandial blood glucose. In our earlier studies the GI value of NFB (apple flavor) was found to be 38.50 (Reddy KK et al., 2009). Peters et al., 1992 showed that Glucerna (70 g carbohydrates/L) in type-1 diabetes patients produced a decrease in basal glucose in comparison with Enrich and Ensure HN supplements, which contain 146 and 141 g of carbohydrates/L respectively. Similarly, Tsai et al., 1987 in their study shown that the

supplementation of 10 g of soy polysaccharides (low GI value) showed a diminished postprandial glucose with no change in insulin in type-2 diabetes patients. The carbohydrate content of the Naturo fruit bar (15.73 g) was relatively lower when compared to HN supplements, which could be a key determinant in altering post-prandial glucose levels. A study (Jayagopal et al., 2002) showed that nutritional supplementation of soy protein (containing 132 mg/day isoflavones) reduced post-prandial glucose in post-menopausal women with type-2 diabetes. Similar to this study we also observed decreased post-prandial blood glucose in diabetic patients consuming Naturo fruit bars for 12 weeks. However, we observed no significant decrease in HbA1C levels. Future studies with large population size may explain the significance of this observation. Previous studies of Gonza´lez-Ortiz et al, (2006) using Glucerna-SR showed that administration of 75 g of glucose decreased the blood glucose and insulin concentration in addition to the total insulin secretion with an increment in the insulin sensitivity in healthy volunteers. The results from our study are in agreement with the above studies, in that a 12week supplementation of a Naturo fruit bar (containing 15.73 g of carbohydrate, 65.48 calories and 1.55 g of dietary fiber) reduced the fasting and post-prandial blood glucose levels (p < 0.05). Nutritional supplementation with 28 % dietary fiber has been shown to reduce total cholesterol and LDL levels (Brown L et al., 1999). In the present study, supplementation of NFB did not affect blood lipids (Cholesterol, HDL cholesterol, LDL cholesterol and Triglycerides) and obesity in type-2 diabetes patients. It is possible that intake of NFB for longer-term (more than 12 weeks) may have some influence on insulin and HbA1C. CONCLUSION In conclusion, the study demonstrates that consumption of NFB (Apple flavor without added sugar) does not increase blood glucose

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levels and may have beneficial effects and may be used as a substitute for healthy food in type2 diabetes patients without any side-effects. We hope that studies like this on indigenously developed products would offer high benefit to the increasing number of diabetes patients (~40 million) in India.

ACKNOWLEDGMENT The authors gratefully acknowledge the financial support by Naturo Food and Fruit Products Pvt. Ltd, Bangalore, India. We thank Prof K.V.S Sharma, Department of Statistics S.V University, Tirupati for his critical input in plotting data as figure.1.

REFERENCES American Diabetes Association Nutrition principles and recommendations in diabetes (2004). Diabetes Care. 27: S36–S46. Brown L, Rosner B, and Willett WW, Sacks FM (1999). Cholesterol-lowering effects of dietary fiber: a meta-analysis. Am J Clin Nutr. 69: 30–42. Fix BM, Low W, Cockram DW (2000). Effects of a liquid nutritional supplement containing a novel carbohydrate system on glucose tolerance in subjects with type 2 diabetes [Abstract]. Ann Nutr Metab. 45(Suppl.1): 277. Friedewald. W.T., Levy,R.I. and Friedrickson,D.S. (1972). Estimation of the concentration of Low density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge, Clin.Chem.18, 499– 502. Gonza´lez-Ortiz M, Martı´nez-Abundis E, Herna´ndez-Salazar E, Kam-Ramos AM and Robles-Cervantes JA (2006). Effect of a nutritional liquid supplement designed for the patient with diabetes mellitus (Glucerna SR) on the postprandial glucose state, insulin secretion and insulin sensitivity in healthy subjects. Diabetes Obesity and Metabolism. 8:331–335. Jayagopal V, Albertazzi P, Kilpatrick ES, Howarth EM, Jennings PE (2002). Beneficial effects of soy phytoestrogen

intake in postmenopausal women with type 2 diabetes. Diabetes Care. 25:1709–1714. Jeyaseelan L, Rao PSS (1989). Methods of determining sample sizes in clinical trial. Indian Pediatr. 26: 115–121. Kaur J, Singh P, Sowers JR (2002). Diabetes and cardiovascular diseases. Am J Ther. 9: 510–515. King H, Aubert RE, Herman WH (1998). Global burden of diabetes, 1995–2025: prevalence, numerical estimates, and projections. Diabetes Care. 21: 1414– 1431. Mohan V, Deepa M (2010). Prevalence of diabetes and metabolic syndrome among Asians. Int J Diab Dev Ctries. 30:173–175. Peters AL, Davidson MB, Isaac RM (1989). Lack of glucose elevation after simulated tube feeding with a low carbohydrate, high-fat enteral formula in patients with type 1 diabetes. Am J Med. 87: 178–182. Peters AL, Davison MB (1992). Effects of various enteral feeding products on postprandial blood glucose response in patients with type I diabetes. PEN 1992; 16: 69–74. Reddy

KK, Nagaraju I, Giriprasad B, Lakshminarayanan G, Chandra Sekhar R, Damodar Reddy C (2009). Determination of Glycemic Index Value

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of Naturo Fruit Bars. J Physicians India. 57:748–750.

Assoc

Rose GA, Blackburn HG, Gillum RF, Prineas RJ (1982). Cardiovascular survey methods. WHO Publication, 2nd Edn, No. 56. Geneva: World Health Organization. Shaw J.E, Sicree, R.A, Zimmet P.Z (2010). Global estimates of the prevalence of diabetes for 2010 and 2030. Diab Res and Clin Practice, 87: 4–14. Singh RB, Ghosh S, Niaz MA, Rastogi V (1997). Validation of physical activity and socioeconomic status questionnaire in relation to food intakes for the five city study and proposed classifications for Indians. Assoc Physicians India. 45: 603–607.

Source of Support: Nil

Tsai AC, Vinik AI, Lasichak A (1987). Effects of soy polysaccharide on postprandial plasma glucose, insulin, glucagon, pancreatic polypeptide, somatostatin and triglyceride in obese diabetic patients. Am J Clin Nutr. 45: 596–601. Vanlandingham (1981). patients feeding.

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Wasir JS and Misra A (2004). The Metabolic Syndrome in Asian Indians: Impact of Nutritional and Socio-economic Transition in India. Metabolic Syndrome and Related Disorders. 2: 14–23. Weiner JS and Lourie JA (1981). General medical examinations, Techniques. Practical Human Biology. Academic press, London.

Conflict of Interest: None Declared

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Original Research Article HYPOGLYCEMIC EFFECTS OF AQUEOUS AND ETHANOLIC EXTRACTS OF DANDELION (TARAXACUM OFFICINALE F.H. WIGG.) LEAVES AND ROOTS ON STREPTOZOTOCIN-INDUCED ALBINO RATS Chinaka Nnamdi C1*, Uwakwe A. A2. and Chuku L. C3. 1, 2, 3

Department of Biochemistry, University of Port Harcourt, Rivers State, Nigeria. *Corresponding author: E-mail: cn_chinaka@yahoo.com, (+234)8039397700, (+234)8027205705. Received: 06/04/2012; Revised: 20/05/2012; Accepted: 31/05/2012

ABSTRACT The effects of aqueous and ethanolic extracts of Taraxacum officinale F.H. Wigg. leaves and roots on fasting blood glucose (FBG) levels of normal and streptozotocin-induced diabetic Wistar albino rats (Rattus rattus) were studied. Exactly 75 Wistar albino rats weighing between 100–225 g were used for the study, and a total of four groups were created. Two groups were divided into six sub-groups of five rats each for the leaf and root extracts respectively, with the remaining two groups being the normal control rats (NCR) and diabetic control rats (DCR). The two sub-groups were thus; sub I, comprising of sub-groups 1–4 which were for diabetic test rats (DTR) on 6% and 10% of aqueous and ethanolic extracts of leaves respectively, while sub-group 5 and 6 were normal test rats (NTR) on 10% of both extracts of leaves respectively. Same was applicable for sub II which represents the roots extracts. After STZ-induction, the course of hyperglycemia was monitored by estimation of FBG. Then administration of T. officinale leaf and root extracts (Aq. and Et.) commenced and lasted for 21 days. Changes in FBG concentration between the NCR and DCR against NTR and DTR treated with two doses (300 mg/kg and 500 mg/kg bodyweight twice a day) of the extracts were evaluated using one way Analysis of Variance (ANOVA). When compared, the FBG levels of the DCR and DTR varied significantly (P < 0.05). Whereas the mean conc. of FBG levels of NCR was (4.4 ± 0.12 mmol/l) that of DCR was (27.1 ± 1.59 mmol/l). On administration of 6% and 10% concentration of the extracts to the DTR and NTR, statistically significant decrease (P ≤ 0.05) in glucose concentration was observed. The FBG level of DTR on 6% leaf extract dropped from 17.1 ± 0.18–9.3 ± 1.86 mmol/l, those on 10% dropped from 18.3 ± 2.58– 9.9 ± 1.00 mmol/l. The NTR on 10% leaf extract dropped from 9.9 ± 0.76–7.9 ± 1.00 mmol/l. For the root extract, FBG levels of DTR on 6% and 10% extracts dropped from 15.8 ± 0.18– 9.6 ± 2.10 mmol/l and 16.0 ± 0.71–7.5 ± 1.46 mmol/l respectively. The NTR on 10% roots extracts fell from 7.55 ± 1.61–3.4 ± 1.23 mmol/l. The results of this study strongly suggest that although T. officinale leaves and roots possess hypoglycemic properties, the roots of ethanolic extraction are relatively more potent and may be beneficial in the management of diabetes.

Keywords: Aqueous (Aq.), Ethanolic (Et.) Extract, Fasting blood glucose (FBG), Hyperglycemia, Hypoglycemia, Streptozotocin (STZ), Taraxacum officinale. Global Journal of Research on Medicinal Plants & Indigenous Medicine


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GJRMI, Volume 1, Issue 6, June 2012, 211–217

INTRODUCTION

The use of plants as a natural cure for several ailments and diseases is one of the oldest practices by mankind. Hence the need to study medicinal herbs and the knowledge of their uses are imperative over the years since in treatment of disease, it is not the drug used to treat and cure the disease that matters, but while doing this, the conservation of the body organisms is the paramount issue (Robert et al., 2007). Humans are always in need of help for their many diseases and illnesses and this help comes from nature. The study and understanding of ethno-botanical information, chemical constituents of plants, and the probable therapeutic applications of the plant drug help in improving health problems. Plants play important roles as vessels with chemical constituents that possess pharmacological potentials (Maurice 2003). Taraxacum officinale F. H. Wigg., commonly known as Dandelion (from the French dent-de-lion meaning lion’s tooth) is thought to have evolved about thirty million years ago in Eurasia. They have been used by humans as food and as a herb for much of recorded history (Dijk et al., 2003). It is an herbaceous perennial plant of the family Asteraceae (compositae), and two major species, T. officinale and T. enythrospermum, are found as weed worldwide. Both species are edible in their entirety. Like other members of the Asteraceae family, they have very small flowers which are yellowish to orange yellow in colour collected together into a compositae (flower head). Each single flower in a head is called floret and they number 40 to over 100 per head. They grow generally unbranched taproots, producing one to more than ten stems that are typically 5–40 cm tall and sometimes up to 70 cm tall. The leaf margins are typically shallowly lobed to deeply lobed and often lacerate or toothed with sharp or dull teeth. The chief constituents of dandelion root are taraxacin, taraxacerin, and inulin (a sort of

sugar which replaces starch in many of the Dandelion family, Asteraceae), gluten, gum and potash. Dandelions are one of nature’s richest green vegetable sources of betacarotene, from which vitamin A is created (14 000 iµ/100 g leaves vs. 11 000 iµ/100 g in carrots). It is a valuable herb and extremely versatile, as the whole plant can be used for medicinal as well as culinary purposes. Medicinally, dandelion has been considered to be an aperient, diuretic, stimulant, stomachic, tonic, anti-diabetic and detoxicant (Clarke 1997). Forty percent of the mature root is inulin, a mixture of complex carbohydrates known as fructo-oligosaccharides (FOS). Based on clinical studies, intake of FOS significantly increases beneficial bifido-bacteria within the gastrointestinal tract and eliminates pathogens. FOS also stimulates the immune system, increases mineral absorption and suppresses abnormal cell growth. The high levels of FOS in dandelion root and its water extract also help to keep blood sugar levels constant and reduce hyperglycemia (Yashpal 2004). 2.0

MATERIALS AND METHODS

2.1.0 Plant Source and Identification The plant Dandelion (Taraxacum officinale) leaves and roots were sourced from farm lands at Umuzi and Umudim villages in Umudioka Ancient kingdom, Orlu local government area of Imo State and the species was identified and confirmed by Dr. F. N. Mbagwu of the Department of Plant Sciences and Biotechnology, Imo State University, Owerri, Imo State. 2.1.1 Chemicals and Reagents All chemicals and reagents used were of analytical standard and were obtained from reputable sources. 2.2.0 Preparation and Administration of Streptozotocin (STZ) The range of diabetogenic dose of STZ is quite narrow and a light overdose may cause

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the death of many animals (Lenzen et al., 1996). 5 g of STZ was dissolved in 100 ml of distilled water to give a 5% stock solution of which a single dose of 70 mg/kg body weight was injected intraperitoneally to the rats. 2.2.1 Preparation of Plant Extract Aqueous Extract Fresh leaves and roots of the plant (Taraxacum officinale) were washed with distilled water to remove debris and contaminants, after which they were dried. The leaves and roots were homogenized into fine powder respectively. The aqueous pulverized plant leaves and roots were respectively prepared by weighing out 100 g of pulverized leaves and roots into 1 l of distilled water respectively. The resultant mixture was allowed to stand for 24 h with occasional shaking after which it was filtered. The filtrate was evaporated and dried to powder with the aid of a thermostatic water bath at a temperature of 50°C. An aliquot of the extract was prepared by dissolving 6 g in 50 ml and 10 g in 50 ml of distilled water respectively to form the two concentrations which served as stock crude drug and stored at 4°C. Ethanolic Extract Fresh leaves of the plant (T. officinale) were washed with distilled water to remove debris and contaminants, after which they were dried. The leaves and roots were homogenized into fine powder respectively. 100 g of powdered leaves and roots were soaked respectively in 500 ml of absolute ethanol and the resultant mixture was allowed to stand for 24 h with occasional shaking, after which it was filtered. The filtrate was evaporated with rotary evaporator and dried to powder with the aid of a thermostatic water bath at 45°C. 2.3 Extract Administration The test rats were administered 300 mg/kg and 500 mg/kg body weight of concentrations

of aqueous and ethanolic leaves and roots respectively twice daily using a gavage via intubation for 21 days, according to the experimental plan/grouping. 6% aqueous extract of leaves and roots were prepared respectively by weighing 6 g of the various extracts (aqueous and ethanolic) and dissolved in 50 ml of water. [6000 mg in 50 ml (3000 mg in 25 ml)] Each rat was administered 0.5 ml (e.g. 200 g rat) of the solution via intubation twice daily for 21 days. 10% aqueous extract of leaves and roots were prepared respectively by weighing 10 g of the various extracts (aqueous and ethanolic) and dissolving in 50 ml of distilled water. [10000 mg in 50 ml (5000 mg in 25 ml)] Each rat was administered 0.5 ml (e.g. 200 g rat) of the solution via intubation twice daily for 21 days. The mode of administration and treatment of the animals according to their experimental regimen/groups is shown in the Table 2.0 2.4 METHOD OF BLOOD COLLECTION Blood used for analysis was collected via the tail vein by dilating the tail veins with methylated spirit and xylene after which the tip of the tail is cut off and analysis done immediately with the blood using an automated Accu chek glucometer and strips for fasting blood glucose. 2.4.1 ASSAY METHOD 2.4.2 Fasting Blood Glucose (FBG) Glucose concentration was determined after an enzymatic oxidation in the presence of glucose oxidase. The hydrogen peroxide formed reacts under catalysis of peroxide with phenol and 4aminophenazone to form a red violet quinoneimine dye as an indicator which is measured glucometrically and the results are expressed in mmol/l.

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www.gjrmi.com Reaction Principle; Glucose + O2 + H2O

GoD

gluconic acid + H2O2 PoD

2H2O2 + 4-aminophenazone + phenol

quinonemine + 4H2O.

Table 2.0

Feeding illustration

GROUPS 1

2

3

4

5

6

7

8

9

10

11

12

13

14

5

5

5

5

5

5

5

5

5

5

5

5

5

5

Feed + water

+

+

+

+

+

+

+

+

+

+

+

+

+

+

STZ(70 mg/kg) 0.2 ml

-

+

+

+

+

+

+

+

+

+

Aq. leaves extract 300 mg/kg

+

Aq. leaves extract 500 mg/kg

+

+

Et. leaves extract 300 mg/kg

+

Et. leaves extract 500 mg/kg

+

+

Aq. roots extract 300 mg/kg

+

Aq. roots extract 500 mg/kg

+

+

Et. roots extract 300 mg/kg

+

Et. roots extract 500 mg/kg − − − Key: + Indicates that item was administered; – Indicates that item was not administered.

+

+

TREATMENT No. of rats per group

3.0 RESULTS AND DISCUSSION Table 3.2.1 FBG values (in mmol/l) of normal and diabetic controls compared to diabetic test and normal test rats treated with aqueous and ethanolic extracts of T. officinale leaves. WEEK 1

2

3

4.5 ± 0.47a 23.5 ± 0.87c 14.2 ± 2.01b 12.9 ± 2.85b 13.6 ± 2.19b 13.4 ± 3.39b 6.7 ± 0.50a 13.1 ± 1.01b

4.5 ± 0.74a 24.1 ± 3.00c 13.9 ± 1.59b 12.6 ± 2.98b 10.6 ± 2.53ab 10.8 ± 1.21ab 6.0 ± 0.55a 12.2 ± 0.79b

4.3 ± 0.18a 33.6 ± 0.91d 13.3 ± 1.83b 12.1 ± 2.96b 9.3 ± 1.86a 9.9 ± 1.00ab 5.0 ± 0.84a 10.7 ± 1.16ab

GROUP NCR DCR DTR on 6% Aq. Extract DTR on 10% Aq. Extract DTR on 6% Et. Extract DTR on 10% Et. Extract NTR on 10% Aq. Extract NTR on 10% Et. Extract

Results are Means Standard Deviation of triplicate determinations. Values in the same column with different superscripts letters are statistically significantly at 95% confidence level (P ≤ 0.05).

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Table 3.2.2 FBG values (in mmol/l) of normal and diabetic controls compared to diabetic test and normal test rats treated with aqueous and ethanolic extracts of T. officinale roots. WEEK 1

2

3

4.5 ± 0.47a 23.5 ± 0.87c 12.7 ± 3.96b 9.8 ± 1.98b 9.2 ± 1.02a 8.4 ± 0.46a 9.7 ± 2.47ab 5.4 ± 0.74a

4.5 ± 0.74a 24.1 ± 3.00c 12.6 ± 3.73b 10.0 ± 2.30ab 8.1 ± 0.59a 6.8 ± 0.75a 8.5 ± 2.35a 4.1 ± 1.18a

4.3 ± 0.18a 33.6 ± 0.91d 12.0 ± 3.86b 9.4 ± 2.08a 7.2 ± 0.25a 5.5 ± 0.83a 6.2 ± 0.70a 3.4 ± 1.23a

GROUP NCR DCR DTR on 6% Aq. Extract DTR on 10% Aq. Extract DTR on 6% Et. Extract DTR on 10% Et. Extract NTR on 10% Aq. Extract NTR on 10% Et. Extract

Results are Means ± Standard Deviation of triplicate determinations. Values in the same column with different superscripts letters are statistically significantly at 95% confidence level (P ≤ 0.05). DISCUSSION The success recorded in the use of streptozotocin (STZ) for the induction of diabetes mellitus through the administration of 70 mg/kg body weight can be attributed to the work of Ferreira et al., 2002. This achievement was confirmed by evaluation of fasting blood glucose concentration. Experimental rats having blood glucose concentrations above 10 mmol/l were considered diabetic as this represents the threshold (Al-Awadi et al., 1991). Normal control rats maintained a fairly stable level of glucose throughout the study period with a mean value of 4.4 ± 0.12 mmol/l. There was however sustained rise in the level of glucose concentration for the diabetic control rats reaching a hyperglycemic mean level of 33.6 ± 0.91 mmol/l on the last day of the study i.e. third week (see Table 3.2.1). On administration of Taraxacum officinale leaf and roots extracts (aqueous and ethanolic), on the normal treated rats, their fasting blood glucose levels dropped, indicating that the plant especially the root extract has the potency to keep blood glucose levels at normal concentration. However, on administration of the 6% and 10% of the leaves and roots

extracts, there was a remarkable decrease (P ≤ 0.05) in the glucose concentration (especially those treated with 10% ethanolic root extracts) over the period of study (see Tables 3.2.1 and 3.2.2). A major effect of streptozotocin on the pancreatic system was observed and this helped in maintaining a steady increase on the glucose level of the diabetic control rats. Since the glucose level of the diabetic treated rats dropped over the period, when compared to that of the diabetic control rats, it gives credence and suggests a possible β-cell recovery (Okamoto 1970). Normal control rats given water had no significant change (P > 0.05) in the fasting blood glucose levels, while the normal test rats following administration of the leaves and roots extracts (aqueous and ethanolic) showed significant decline in fasting blood glucose. It has been suggested that tannins may lower the rate at which starch is digested and hence blood glucose level by the same mechanism that makes them anti-nutrients. It is also known that tannins may bind directly with pancreatic amylase (the principal enzyme in starch digestion) thus inactivating it. Tannins may also bind with calcium which is needed to stabilize amylase activity or with starch to

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influence its degree of gelatinization or its accessibility to the digestive enzymes (Thompson 1993). In summary, this study has provided some evidence for the hypoglycemic effects of Dandelion (Taraxacum officinale) leaves, compared to its roots and the most effective mode of extraction (aqueous or ethanolic) as well as the most effective dosage to be used in terms of drug other than food, in the management of diabetes mellitus. 4.0

CONCLUSION

This study has provided comparative evidence of the hypoglycemic effects of Taraxacum officinale leaves and roots of

aqueous and ethanolic extraction, on normal rats and streptozotocin-induced diabetic rats. It has shown that the extracts (especially the ethanolic extract) has the potential to improve carbohydrate metabolism. It also indicates that ethanolic extraction of this plant is most suitable compared to aqueous and the roots are more effective compared to the leaves in the management and treatment of diabetes. The experimental findings also indicate that Taraxacum officinale extracts is dose dependent. ACKNOWLEDGEMENT The authors acknowledge support from the World Bank and the Federal Republic of Nigeria under the World Bank step B project.

REFERENCES Al-Awadi, F., Fatana, H. and Shamte, U. (1991). The effect of a plant mixture extract on liver gluconeogenesis in STZ-induced rats. Diabetes Res. 18:163–168. Annapurna, A., Kanaka, Mahalakshimi, D., Murali Krishna, K. (2001). AntiDiabetic activity of a polyherbal preparation (Tinture of Punchparna) in normal and diabetic Rats. Ino Exp. Biol. 39:500–5002. Bailey, C.C. (1947). Streptozotocin diabetes. In Joslin E.P., Root, H.F., White, P. and Mark, A. (eds), The treatment of diabetes, 8th ed. Lea and Febiger, Philadelphia. pp 178–192. Bailey, C.C. and Bailey, O.T. (1943). The production of diabetes mellitus in rabbits with streptozotocin .J. Amer. Med. Ass. 21:1165. Bhattacharya, S.K., Satyan, K.S., Chakibarti, A. (2001). Effects of Trasina on Ayurvedic herbal formulation of pancreatic islet superoxide dismutase activity in hyperglycemia. Indian J. Exp. Biol. 35:297–299.

Clarke, C. B. (1997). Edible and useful plants of California. Berkeley: University of California press. pp 191. Dijk, P. J. Van. (2003). “Ecological and evolutionary opportunities of apomixes: Insights from taraxacum and chondrilla” Philosophical transactions of the Royal Society. Biol. Scie. 358 (1434):1113. Duff, G.L. (1945). The pathology of the pancreas in experimental diabetes. Am. J. Med. Sci. 210:381–397. Elder, C. (2004). Ayuveda for diabetes mellitus. A review of the biomedical literature alternative health med. 10:44–50. Fajan, S.S. and Vinik, A. I. (1989). Insulinproducing islet cell tumors. Endocrinol. Metab. Clin. North Am. 18:45–74. Gray, A. M. and Flatt, P. R. (1997). Pancreatic and extra-pancreatic effects of the traditional antidiabetic plant medica sativa (Lucerne). Br. J. Nutr. 78(2):325–334.

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Gray, A. M. and Flatt, P. R. (1998). Insulinsecretion activity of traditional and antidiabetic plant viscum album Dandelion. Journal of Endocrin. 160:207–212. George, D. and Pamplona-Roger, M. D. (1999). Encyclopedia of medical plants. pp.246–7, 227. Harborne, J. B. (1998). Phytochemical method. A guide to modern techniques of plant analysis (3rd edition). Http://akweeds.uaa.alaska.edu Jia, W., Gao, W.Y., Xiao, P.G. (2009). Autodiabetic drugs of plant origin used in China composite pharmacology and hypoglycemic mechanism. Zhongguo Zhong Yaoza Zhi; 2:113. Lenzen, S., Tiedge, M., Jones, A., Munday, R. (1996). Alloxan derivatives a tool for elucidation of the mechanism of the diabetogenic action of alloxan; In lesson for animal diabetes. E. Shafrir (ed.). Boston Birkhauser. pp. 113–122. Maurice, A. E. (2003). Some common locations of Dandelion in Africa. African handbooks: Volume 12. Mordes, J.P. and Rossini, A.A. (1981). Animal models of diabetes. Am. J. Med: 70:353–360. National Institute of Health (1986). Diet and exercise in non-insulin dependent diabetes mellitus. Concensus development conference statement Vol. 6 No 8. Okamoto, K. (1970). Experimental production of diabetes in: Ellenberg, M. and Rifkin, H. (eds.). Diabetes mellitus: theory and practice, Blackiston publication, Mc Graw-Hill Book Company, New York. pp. 230–243. Source of Support: Nil

Omeodu, S. I. (2006). “The effects of aqueous extracts of mistletoe and garlic on serum AST, ALT and ALP of wistar albino rats with CCl4-induced liver damage. Department of Biochemistry, University of Port Harcourt, Nigeria. pp. 2–8. Robert, F., Barnes, C., Jerry, N., Kenneth, J.M., and Michael, C. (2007). Forages: The science of grassland agriculture. WileyBlackwell pp.11. Schutz, K., Carle, R. and Schieber, A. (2006). Taraxacum- A review on its phytochemical and pharmacological profile. J. Ethnopharmacol. 10–11, 107(3):313–323. Shanmugasanderam, E.R., Gopith, K.L., Radha, S.K. and Rajendram, V.M. (2000). Possible regeneration of the islets of Langerhans in streptozotocin diabetic rats given gymneama sylvestere leaf extracts. J. Ethnopharn; 30:265–9. Sofowora, A.O. (1982). Medicinal plants and traditional medicine in Africa. John Wiley and Sons. pp. 204–208. Takasaki, M., konoshima, T., Tokuda, K., Arai, Y., Shiojima, K. and Agate, H. (1999). Anti-carcinogenic activity of Taraxacum plant. I and II. Biol. Pharm. Bull. 22(6):602–5; 606–10. Thompson, L.U. (1993). Potential health benefits and problems associated with nutrients in foods. Food Res. Inter. 26:131–149. WHO study group. Diabetes Mellitus (1996). WHO Tech. Report Ser. No 727. Yashpal, P.C. (2004). Chemical constituents of dandelion. pp. 12–14.

Conflict of Interest: None Declared

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Original Research Article ANTIBACTERIAL EVALUATION OF ETHANOLIC EXTRACT OF CYNODON DACTYLON (L.) PERS. Kashyap Pranita1*, Sawarkar Hemant A2, Mishra Koushlesh K3 1, 2, 3

Dept. of Pharmacognosy, Shri Rawatpura Sarkar Institute of Pharmacy, Behind Holiday Resort, Kumhari â&#x20AC;&#x201C; 490024, (C.G.), India. *Corresponding author: Ph. No.- 09039823698

Received: 30/03/2012; Revised: 20/04/2012; Accepted: 26/05/2012

ABSTRACT Cynodon dactylon (L.) Pers. of the family Poaceae is extensively used in clinical practice and has reported various pharmacological activities with persistent findings till date. It is a very familiar plant of our surroundings and almost available in all corners of the world. In ethno-medicinal practices, the juice of the plant is used as an astringent and is applied to fresh cuts and wounds. It is used internally in the treatment of chronic diarrhoea and dysentery. It is also useful in the treatment of catarrhal ophthalmia. The leaves of C. dactylon (L.) Pers. are also used in the treatment of hysteria, epilepsy and insanity. In this experimentation, first the whole plant was subjected to ethanolic extraction and then phytochemical screening of the extract was performed for primary & secondary metabolites. The in-vitro evaluation of antibacterial activity was done against E. coli, S. aureus & S. pyogenes. 10% concentration of extract was found to be most effective as antibacterial when compared to other concentrations, standards & controls used in this study.

Key words: Cynodon dactylon, Ethanolic extract, S. aureus, S. pyogenes, E. coli

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INTRODUCTION: Reported in the literature, according to an estimation of the World Health Organization, about 80 percent of the world’s population relies on herbs for its primary healthcare needs. More than 35,000 plant species are being used around the world for the medicinal purposes in traditional and ethnomedicinal practices (Duke, J. (1983). Currently, there is a renewed global interest in the study and use of medicinal plants because such investigations provide important, new lead on novel, active molecules of therapeutic importance. For this reason traditional plant based remedies are back and find increasing application as source of direct therapeutic agents. Considerable scientific data have been generated worldwide in this direction and there is a spurt on natural product study. Based on this rationale, the study has been compiled and provides data on potential medicinal properties, action and uses of plants in order to contribute material, leading to the discovery of new drugs of natural origin (Khosoo,T.N., 1999). Among numerous species of plants growing in the wild in India, Doob Ghas, or Durva or taxonomically Cynodon dactylon (L.) Pers., family Poaceae occupies its unique place and key position in ethnomedicinal practices and traditional medical (Ayurvedic, Unani, Nepalese, and Chinese) knowledge systems. The herbal preparations of this grass are being based on folklore and traditional wisdom (Mishra, M.P.,2006). Literature survey also reveals that fresh leaf’s paste of whole plant is beneficial in the treatment of wounds & infections (http://botanical.com/site/column_poudhia/111 doobi.html, 12 sep 2010). Although the antibacterial activities of hydroalcoholic extract of the whole plant have been reported, no systematic study has been reported on the ethanolic extract of the plant. Hence the present investigation deals with evaluation of antibacterial activity of the whole plant, ethanolic extracts of C. dactylon (L.) Pers. against S. aureus, S. pyogenes, E. coli, three causative organisms well known for their infections caused in Human beings.

Chemical Composition of C. dactylon (L.) Pers. It contains essential oil triticin 12.4%. The other chemical constituents are glycosides, saponins, tannins, flavonoids, & carbohydrates (Basu & Kirtikar 2000). It also contains agropyrene, arunodin, furfural, furfural alcohol, β-ionine, 2-(4’hydroxy phenyl) propionic acid, 2-(3’methoxy-4’hydroxy-phenyl) propionic acid, 3-methoxy-4-hydroxy benzoic acid, phytol, β-sitosterol-D-glucoside, stigmasterol acetate, phagostimulant phytone (6,10-14trimethyl pentadecane-2-one) (Mukherjee, Pulok K., 2002). Cuticular wax contains triacontane, docosanol. tetracosanol, hexacosanol, octacosanol, eicosanic acid & docosanoic acid (spencer & chapmann) (The wealth of India 2nd supplement series). MATERIALS AND METHODS: Collection of Plant material: The whole plant of Cynodon dactylon (L.) Pers. were collected from the Botanical garden of Shri Rawatpura Sarkar Institute of Pharmacy, Kumhari, Dist.- Durg, Chhattisgarh, India in September 2010, cleaned and dried at room temperature in shade and away from direct sunlight. The plant was authenticated by Dr. Mrs. Ranjana Shrivastava, HOD, Dept. of Botany, Govt. V.Y.T. P.G. Autonomous College, Durg (C.G.), by comparing morphological features and a sample voucher specimen of the plant was deposited for future reference. Preparation of Extract The whole plant of C. dactylon (L.) Pers. were collected and dried in the shade and then pulverized in a grinder. The powdered drug was utilized for extraction. Material was sieved with 120 mesh to remove fine powders and coarse powder was used for extraction. A method (described in Mukherjee) was used for extraction of powdered material. Extraction was done by using solvent Ethanol.

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Table 1: Results of Antibacterial activity against E.coli. Zone of inhibition (mm) Extract against E.coli 1

2

3

4

5

Mean

S.D

Result

F-I: 5 mg/ml

13

14

14.5

14

13.5

13.8

0.3

13.8 ± 0.3

F-II: 7.5 mg/ml

15

16.5

16

15

16

15.7

0.5

15.7 ± 0.5

F-III: 10 mg/ml

18.5

17

18.5

17.5

17

17.7

0.2

17.7 ± 0.2

Standard

14.5

15

15.5

14

15

14.8

0.4

14.8 ± 0.4

Water (−ve control)

10

10

10

10

10

10

0

No activity

M.P + P.P

16

17

16.5

16

15.5

16.2

0.7

16.2 ± 0.7

(+ve control)

Table 2: Results of Antibacterial activity against S.aureus. Zone of inhibition (mm) Extract against S.aureus 1

2

3

4

5

Mean

S.D

Result

F-I: 5 mg/ml

14

15

14.5

15

13

14.3

0.4

14.3 ± 0.4

F-II: 7.5 mg/ml

16

16.5

15

16.5

16

16.1

0.2

16.1 ± 0.2

F-III: 10 mg/ml

17.5

18

17

18

17

17.5

0

17.5 ± 0

Standard

15

14.5

16

15.5

15

15.2

0.3

15.2 ± 0.3

Water (−ve control)

10

10

10

10

10

10

0

No activity

M.P + P.P

16

17

16.5

17

16.5

16.6

0.5

16.6 ± 0.5

(+ve control)

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Table 3: Results of Antibacterial activity against S.pyogenes. Zone of inhibition (mm) Extract against S.pyogenes 1

2

3

4

5

Mean

S.D

Result

F-I: 5 mg/ml

13

12.5

13.5

14

13.5

13.3

0.3

13.3 ± 0.3

F-II: 7.5 mg/ml

14.5

16

15

16

15

15.3

0.4

15.3 ± 0.4

F-III: 10 mg/ml

17

17.5

16

17

16

16.7

0.6

16.7 ± 0.6

Standard

14

14.5

13.5

14.5

13.5

14.0

0.4

14.0 ± 0.4

Water (−ve control)

10

10

10

10

10

10

0

No activity

M.P + P.P

15

14

16

15

15

15

0.5

15 ± 0.5

(+ve control) Diameter of hole (well) - 10mm; S.D - Standard Deviation. Deviation

Figure 1 (a&b); Antibacterial activity of Ethanolic extract against Escherichia coli.

1 (a)

1 (b)

Preliminary Phytochemical Screening Phytochemical screening was carried out according to standard methods. Extracts shows the presence of carbohydrates, glycosides, flavonoids, saponins, alkaloids, phenolic compounds, tannins, fixed oil, & mucilage (Wallis T. E., 2000). ). Inorganic constituents present in selected plant C. dactylon are calcium, magnesium, chloride, iron & ssulphur (Trease And Evans., 2002).

Thin layer chromatography of various extracts of C. dactylon has confirmed the presencee of Secondary metabolites like: alkaloids, glycosides, saponins, tannins & flavonoids as the colour of spots & their Rf value of standard (Ayurvedic Pharmacopeia of India) matches to that of sample (Khandelwal 2005).

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Evaluation of Antimicrobial Activity The in-vitro antibacterial activity of Cynodon dactylon (L.) Pers. extract was carried out by using the cup plate: Agar well diffusion method (Acumedia, Neogen corporation, google.com). All the extracts were separately dissolved in ethanolic medium in 3 dif different concentrations 5 mg/ml, 7.5 mg/ml, and 10 mg/ml solutions. Povidone-Iodine Iodine (5 mg/ml) was used as standard antibacterial agent. The antibacterial activity was investigated by using 3 different microorganisms (bacteria), gram negative (E. coli), gram positive ((S. aureus), and haemolytic microorganism ((S. pyogenes). For antibacterial activity 6 groups were made, in which first group was standard: (Cipladine) Povidone-iodine, iodine, second group was positive control: Methyl Paraben + Propyl Paraben (1:2), third group was negative control: Water and the rest 3 test groups were: Formulation Formulation-I,

Formulation-II, II, Formulation-III. Formulation Antibacterial activity was found satisfactorily in all the 3 microorganisms. The antibacterial activity was evaluated by employing 24 h cultures of Escherichia coli using MacConkey Agar media (Biokar diagnostics, biokardiagnostics.com), Staphylococcus aureus using Nutrient Agar media and Streptococcus pyogenes using Blood Agar Base se media (www.biosynth.com). 10 mm bore was made in a petridish. The microorganism inoculated plates were maintained ained at room temperature for 2 h to allow the diffusion of solution into the medium. The petri dishes used for antibacterial screening for E. coli was as incubated at 36+10 for 24 h (Ruoff K.L. et al., 2003), 2003 for S. aureus at 37+10 for 7 days and for S. pyogenes incubated at 37+10 for 3 days. The diameters of zone of inhibition surrounding each of the wells were recorded. (Bose and Bose 2008).

Figure 2 (a&b): Antibacterial activity of Ethanolic extract against Staphylococcus aureus.

2 (a) Figure 3 (a&b):

2(b)

Antibacterial activity of Ethanolic extract against Streptococcus pyogenes.

3 (a)

3 (b)

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RESULTS & DISCUSSION The antibacterial results reveals that [Table 1, 2, and 3] the activity of the crude extract of C. dactylon plant is encouraging. Antibacterial activity was done by using Agar well diffusion method; Povidone-Iodine was used as standard for comparing results for antibacterial activity. The zone of inhibition for E. coli of F-I: 5 mg/ml is 13.8 mm, F-II: 7.5 mg/ml is 15.7 mm and F-III: 10 mg/ml has 17.7 mm, for S. aureus of F-I: 5 mg/ml is 14.3 mm, F-II: 7.5 mg/ml is 16.1 mm and F-III: 10 mg/ml has 17.5 mm, and for S. pyogenes of F-I: 5 mg/ml is 13.3 mm, F-II: 7.5 mg/ml is 15.3 mm and FIII: 10 mg/ml has 16.7 mm. The zone of inhibition of standard drug Povidone-iodine has 14.8 mm for E. coli, 15.2 mm for S. aureus and 14 mm for S. pyogenes and of positive control m.p + p.p has 16.2 mm for E.coli, 16.6 mm for S.aureus and 15 mm for S.pyogenes. All the three concentration of extract shows antibacterial activity but 10% of ethanolic extract shows better antibacterial activity against Escherichia coli, Staphylococcus aureus & Streptococcus pyogenes. For antibacterial activity, formulation-III which contains 10% Ethanolic extract of C. dactylon (L.) Pers. has shown maximum activity, against all the three microorganisms which were selected for the activity. The zone of inhibition for E. coli was (17.7 mm), for S. aureus it was (17.5 mm), and for S. pyogenes it was (16.7 mm) which shows the maximum range than

rest of all the concentrations. The zone of inhibition of standard drug Povidone-iodine has between F- I and II for all the microorganisms. On the basis of zone of inhibition Cynodon dactylon (L.) Pers. showed better antibacterial activity when used in concentration of 10 mg/ml against E. coli, S. aureus, and S. pyogenes when compared to standard & positive control. CONCLUSION From the above results, it was concluded that the plant C. dactylon (L.) Pers. showed significant antibacterial activity. The experimental evidence obtained in the laboratory model could provide a rationale for the traditional use of this plant as antibacterial for wound healing. The activity was performed about to the standard laboratory procedures. The plant may be further explored for its phytochemical profile to recognize the active constituent accountable for antibacterial activity. Thus the present experiment from ethanolic extract of C. dactylon (L.) Pers. has been scientifically proved to be beneficial as an antibacterial for specific bacterial infections. ACKNOWLEDGEMENT Authors sincerely thanks to the editorial team for peer review & Dr. Mrs. Pranita Kashyap for her professional expertise guidance and Mr. H. A. Sawarkar for his prompt suggestions & help.

REFERENCES Acumedia, (2008), Neogen corporation PI 7150, Rev 04, October, www.google.com.

Biokar diagnostics, Reu des Quarante Mines, France; www.biokardiagnostics.com.

Aneja

Duke, J. Handbook of Energy Crops, (1983), Internet Publication- Friends of Plants for future species Database, 1997–2000.

KR, (2007), Experiments in Microbiology Plant Pathology and Biotech, 4th edition, New Age international Publisher,

Basu, Kirtikar (2000), Indian Medicinal Plants. 2nd ed. Elsevier publications Vol. 3. p. 1042–43

Internet resources, Traditional medicinal knowledge about useful herb Doobi (Cynodon dactylon) in Chhattisgarh, India.

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GJRMI, Volume 1, Issue 6, June 2012, 218–224 Journal of Pharmaceutical Sciences. Vol. 70 (6), 821–823.

http://botanical.com/site/column_poudh ia/111 doobi.html, 12 sep 2010. Khandelwal K R. (2005) Practical Pharmacognosy Techniques and th Experiments, 19 edition, Nirali Prakashan, Page No. 149–156Khosoo, T. N. (1999), Dharma of Ecology. Current Science.Vol.77 No. 9:1147– 1153. Mishra, M. P. (2006), Succession of fungi and their eco-microbial involvement in the decay of Cynodon dactylon Pers., Ph.D. Thesis: 14–21. Mukherjee, Pulok K. Quality Control of Herbal Drugs 2002. Business Horizon, 5-34, 97, 98, 181–195, 248, 249, 562–567. Ruoff K L, Whiley R A, Beighton D. Streptococcus. In: Manual of Clinical Microbiology. Murray PR, Baron EJ, Jorgensen JH, Pfaller MA, Yolken RH, eds. 8th ed. ASM Press. 2003: 405– 421. Bose S. and Bose Arti . (2008). Antimicrobial activity of Acanthus ilicifolius. Indian

Source of Support: Nil

The

Ayurvedic Pharmacopoeia of India, Government of India Ministry of Health & Family Welfare. 1st ed. Part-1, Vol.II, p. 62.

‘The Wealth of India’, First supplement series (Raw Materials), (2007) Second print, Vol-1: A-Ci, National Institute of Science Communication & Information Resources, CSIR, Page No.17. The wealth of India 2nd supplement series, Raw materials, CSIR, Vol. 1 page no.331– 332 Trease

and Evans (2002), Textbook of Pharmacognosy, Elsevier; 15th ed. p. 204–205, 479

Wallis

T.E. (2000): Text book of Pharmacognosy, Plant profile for Cynodon dactylon (Bermudagrass) USDA Plants.html, 5th ed. Vol.-II, p. 243–244.

Www.biosynth.com/index.asp.topic_id 177 & g 19 & m 254.

Conflict of Interest: None Declared

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Review Article SACRED TREES OF TEMPLES OF TIRUCHIRAPPALLI, TAMIL NADU â&#x20AC;&#x201C; THE NATURAL AND ECOLOGICAL HERITAGE OF INDIA

Umavathi R1, Parvathi A2* 1,2

Department of Botany, Holy Cross College (Autonomous) Tiruchirappalli - 620 002, Tamilnadu, India. *Correponding author E.mail- parvathi.adapa@gmail.com

Received: 30/04/2012; Revised: 31/05/2012; Accepted: 03/06/2012

ABSTRACT Sacred trees (Sthalavrikshas) representing one form of nature worship are common in the temples of Tamil Nadu and other parts of India. Sacred trees (Sthalavrikshas) are important not only for their botanical, economical, medicinal, environmental, religious and mythical values but also forms an important biological heritage of our nation that plays role in the conservation of environment and biodiversity. The present study dealt with the survey of the trees of some temples in and around Tiruchirappalli, Tamil Nadu. The study documented about fifteen plants associated with twenty temples.

Key words: Sacred Trees, Biological uses, Ecological heritage, sthalavrikshas, Tiruchirappalli.

.

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INTRODUCTION Sacred Trees (Sthalavrikshas) have a unique place in the fabric of Indian heritage. Trees symbolize knowledge and spirituality. In ancient times, the trees were worshipped along with Gods and established in the temples as Sthalavrikshas i.e. trees associated with the deity in the temple and become an inseparable part of the faith. Such trees were guarded and their saplings from mother plants were given to other temples. Trees such as Ficus, Neem and Tamarind are the abode trees of spirits and propitiated by tying toy cradles when desiring a child or at the birth of an infant. Similarly, they tie a black cloth sometimes with salt in it to ward off the evil eye. Similarly, yellow and white cloths are tied in fulfillment of a vow. Ayurvedic and Siddha physicians come to the temple market, not only to gather herbs but also to swap information about many medicinal properties. They also conduct clinics during sojourn. Many temples have Sthalavrikshas, as in worship of Shiva, Aegle marmelos (Vilvam) is preferred. Similarly, Ocimum sanctum (Tulasi) is favoured for the worship of Vishnu, while Nymphaea stellata (Lotus) is a popular choice for offering to all deities. The leaves, flowers and roots may also be added to create fragrant garlands and garlands may be wrapped in either leaf or bark of the plantain. The sheath of the leaf base of Areca tree may also be used to pack and transport flower garlands. The common plants in use are Jasmine, oleander, marigold, leafy branches of Tulasi, tender shoots of Marikozhundhu, Vilvam and roots of Vettiver. Sthalavrikshas are valued for their botanical, medicinal, environmental, religious and mythical importance. The Sthalavrikshas of Tamil Nadu constitute a part of genetic resources for the conservation of species diversity. Many living organisms including man depend upon the plants for their food, shelter and medicine. Propagation of Sthalavrikshas in temples contributes to the conservation of our floral diversity. Some trees are important for their economic role in ship building or in the timber industry, some for their produce, some for providing homes for

various animals, birds and others for their medicinal and air purifying qualities. Usually Ocimum sanctum and Aegle marmelos are given to the worshippers after their prayer in Vishnu and Shiva temples respectively. This habit shows characteristically the importance of medicinal plants in Indian System of Medicine. Medicinal parts of the Sacred Trees (Sthalavrikshas) are used in different forms. It is given in the form of paste, juice, dried powder and made into tablets and juices mixed with sugar and honey to cure various diseases. (Trivedi Priya Rajan 1996; Amirthalingam 1998; Jain 2004). Since no serious attempt has been made so far to study the Sacred Trees (Sthalavrikshas) of Tiruchirappalli, the present study was under taken to survey and document the Sacred Trees associated with various temples. MATERIALS AND METHODS Twenty temples of Tiruchirappalli were visited during the period of June 2008 to May 2009 and sixteen Sacred Trees were recorded (Table 1) & (photo slides 1â&#x20AC;&#x201C;4). It was noted that the same species appeared in more than one temple. Herbarium of the Sacred Trees was preserved at the Department of Botany, Holy Cross College (Autonomous), Tiruchirappalli, Tamil Nadu. Tiruchirappalli, an ancient city of the Cauvery delta is one of the greatest cities of south India situated at the very centre of Tamil Nadu, is equidistant from almost all the major cities of the south including Chennai, Bangalore, Tirupati and Trivandrum. It enjoys a strategic importance which is in tune with its age old agriculture pre-eminence, and its ever growing industrial, commercial and educational advancement. Tiruchirappalli not only abounds in temples but also harbours lot of medicinal plant wealth. It is one of the places of historical importance in Tamil Nadu. The temples in and around the town, the Mainguard gate and the church with its tall pinnacle speak of its glorious past. It is a place of pilgrimage for people of all religions. It is encircled by a number of other pilgrim centers like Srirangam, Thiruvani kovil, Samayapuram and Vayalur.

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GJRMI, Volume 1, Issue 6, June 2012, 225â&#x20AC;&#x201C;233 Photo slides 1 & 2

1. Samayapuram â&#x20AC;&#x201C; Mariayamman Kovil- Azadirachta indica,

2. Srirangam - Sri Ranganathar Kovil- Calophyllum inophyllum L

RESULTS AND DISCUSSION The Sacred Trees documented from various temples visited are presented in Table-1 along with the name of the place, temple, deity, binomial and common name. The botanical features, ecological, economic and medicinal potentials were enumerated based on the literature surveyed (Chakraberty 1993; Gamble 1967; Airy Shaw 1973; Ambasta 1986; Bentley Robert & Trimen Henry 2000; Mathew 1983; Sood et al., 2005). Aegle marmelos Corr. (Rutaceae) is a small medium sized thorny tree with a spreading crown. The transpiration of leaves helps in maintaining the humidity of the immediate surroundings. The root system helps in retention of water in the soil. The wood is suitable for making charcoal. The gummy substances in which the seeds remain embedded are used as an adhesive, varnish and in cementing. The yellow dye is obtained from the unripe fruits. A drug balae fructose extracted from the fruits with mucilage and pectin content is very useful for treating chronic diarrhoea, dysentery, hemorrhoids and swellings. The leaf juice mixed with black pepper is used to treat jaundice. The antibiotic

potential of the leaf, fruit and root helps in curing diarrhoea, dysentery, asthma and fever. The bark and root are soaked in water over night in a copper vessel are used to cure blood pressure, diabetes and leprosy (Venkatesan et al., 2009; Rajan et al., 2011). Ficus religiosa L. (Moraceae) is a large tree with well-developed crown and few low buttresses. The tree releases a considerable amount of oxygen and one tree can supply the oxygen requirement of 6 persons per day. It is a suitable plant species to be planted around the industrial area. The smoke of the twigs possesses germicidal and antimicrobial effect that kills the germs of the environment. The twigs of the plant are used in yagnas. Leaves are used as good fodder for elephant and cattle. Wood is used for making packing cases, articles and match sticks. Bark is used in tanning. Fruits and tender leaf buds are eaten. Latex after hardening is used as lac or sealing wax. The juice of the bark is astringent and used as mouth wash for curing toothache and strengthening gums. The juice extracted by soaking pieces of tender bark in water over night is given for excessive urination during jaundice. The dried fruits are pulverized and given to women during child birth as folk remedy (Uma et al., 2009; Inder Kumar Makhija et al., 2010).

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No 1. 2.

3

4 5

6 7

8

9 10

Name of the Name of the temple place Anna Nagar Varasakthi Vinayagar

Deity of the temple Vinayagar &Shivan BHEL Sri Dharmasasta Thiru Shivan & Kovil & arulmigu Murugan Thiru Murugan Edastreet SriSelvaVinayagar Vinayagar Market Kovil Sri Mahakaliyamman Kaliyamman Kovil Gandhi MarKet Sri Kaliashanathar Shivan Thiru Kovil Keelaputhur Sri Bala Vinayagar, Vinayagar Sri Selva Mariyamman Mariayamman Sri Vallitheivanai Theivanai Kovil MalaiKottai Sri Selva Vinayagar Vinayagar Kovil OFT (Ordnance Raja Vinayagar Kovil Vinayagar Factory Tiruchirappalli) Palanganangudi (i)Thanagamani Kovil Thanagamani (ii)Mariayamman Mariayamman Kovil Samayapuram Mariayamman Kovil Mariayamman Sangilianda Puram

11

Srirangam

12

Thiruvanai Kovil

13

Thiruverumbur

14

Thiruppanjali

15

Uttamar Kovil

Name of the Sacred Common name trees (Sthalavrikshas) Aegle marmelos,Corr. Vilva maram Aeglemarmelos,Corr. Ficus religiosa,L.

Vilva maram Arasa maram

Ficus religiosa,L.

Arasa maram

Azadirachta indica,A.Jus s Aegle marmelos, Corr.

Vembu

Musa paradisiaca,L.

Vazhai

Ficus religiosa,L.

Arasa maram

Ficus religiosa,L. Ocimum sanctum,L.

Arasa maram Tulasi

Azadirachta indica,A.Juss Ficus virens,Aiton Azadirachta indica,A.Jus s Sri Selva Kaliyamman Kaliyamman & Ficus religiosa,L. Kovil Vinayagar Azadirachta indica,A.Juss Sri Ranganathar Kovil Ranganthar Calophyllum inophyllum L Jumbukeshwarar & Shivan & Aegle marmelos,Corr. Akilandeshwari Kovil Akilandeshwari Syzygium cumini(L)Skeel s Sri Jumbukeswarar Shivan Aegle marmelos,Corr. Kovil Neelivaneswarar Shivan Aegle marmelos,Corr. Kovil Musa paradisiacal, L. Uttamar Kovil Shivan Aegle marmelos,Corr.

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Vilva maram

Vembu Itchi maram Vembu Arasa maram Vembu Pinnai Vilva maram Naval Vilva maram Vilva maram Vazhai Vila maram


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Anthyodaya Medicinal Sukkiran Farm- Navagraghas (Shukra)

Ficus racemosa,L.

Atti

Chandran (Chandra)

Butea monosperma, (Lamk.)Taubert.

Purasu (purasa maram)

Sevvai (Mangala)

Diospyros ebenum,J. Konig ex Retz.

Thaali

Raahu

Cynodon dactylon ,Pers.

Arugam pilu

Sani

Allmania nodiflora,(L.) R.Br.ex Wight.

Vanni keerai

Kethu Guru

Imperata cylindrical,(L.) Tharpaipullu Raeusch Ficus religiosa,L. Arasa maram

Puthan (Budha)

Achyranthes aspera,L.

Suriyan (Surya)

Calotropis gigantea,(L.) Yerukku Dryand.

Azadirachta indica A. Juss. (Meliaceae) is a large evergreen tree with an irregularly rounded crown. It releases lot of oxygen and purifies the environment. The wood is used for carving the images of the Gods, toys and for making agricultural implements, carts, boards and panels. The mature leaves and seed cake is used to feed cattle to increase the output of milk. The bark is used in preparing a dye for colouring fine textured fabrics. The seeds yield deepest yellow oil used as an antiseptic and used in preparing tooth paste and soap making. Capsules of neem oil have been recommended to treat diabetes. Sodium nimbate obtained from neem oil is used in birth control. Nimbin and Nimbidine obtained from the seeds are

Nayurivi

used in curing skin diseases. Neem oil is also applied as an antiseptic dressing in leprosy, scabies and ring worm (Khan et al., 1999; Amal kumar Ghimeray et al., 2009). Ficus virens Aiton (Moraceae) is a densely foliaceous tree with a crown of equal spread. The plant releases good amount of oxygen and maintains the moisture content of the atmosphere. Fruits are edible and the succulent leaves are relished by animals. Wood and bark are good source of paper pulp. The bark is diaphoretic. Decoction of bark is used in sore throat.

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Calophyllum inophyllum L. (Clusiaceae) is an evergreen tree with sweet scented flowers and the spreading crown. The scent emitted from the flowers kills the germs around. The timber is fairly hard and is used in ships and boat making, railway sleeper and plywood. The oil obtained from the seeds is used for soap making and illumination purposes. The fruit

yields valuable gum. The bark of the tree has an antimicrobial effect. Oil extracted from the seed kernel is used as a remedy for pain in the joints and muscles. The refined oil is injected intramuscularly to relieve pain in leprosy. The flower juice is used to treat nervous disorders and paralysis (Minh Hienhaetar et al., 2009; Uma Shankar Mishra et al., 2010).

Photo Slides 3 & 4 3. Anna Nagar - Varasakthi Vinayagar temple- Aegle marmelos

4. Malai Kottai â&#x20AC;&#x201C; Sri Selva Vinayagar Kovil- Ficus religiosa L

Syzygium cumini (L.) Skeels (Myrtaceae) An evergreen tree with a round crown, white flowers and purple fruits. The tree is pollution tolerant, assimilates more carbon dioxide from the atmosphere and liberates oxygen, balancing the ratio of oxygen and carbon dioxide in the atmosphere. Fruits are edible, good in taste and used in puddings. Wood is hard and durable. Used for building and agricultural purposes. Leaves are used as fodder. Fruit pulp is useful in treating diabetes and the decoction of the bark is used as a mouth wash to treat spongy gums (Jeethu Anu Mathews et al., 2011; Shanbhag et al., 2011).

the lac insects. Leaves are used for making plates and cups. Dried leaves are used as beedi wrappers. Ropes are made out of roots. The coarse fibre of the inner bark is used for boat making. The gum obtained from the tree is rich in gallic and tannic acids. The seeds contain proteolytic and lipolytic enzymes. The bark is astringent and used to treat tumors and menstrual disorders (Ambersing Raj put et al., 2011; Rmanjaneyulu et al., 2011).

Ficus racemosa L. (Moraceae) is a large tree with a well developed crown. It is pollution tolerant and absorbs pollutants from the atmosphere. It is a suitable plant to be planted in the industrial area. Wood is used for making packing cases and house hold articles. Roasted fruits are used as breakfast food. The root is used in diarrhoea and diabetes. Fruits are carminative (Krishna Murti et al., 2011; Poongothai et al.,2011).

Musa paradisiaca L. (Musaceae) is a large herbaceous plant with a pseudo stem and a globose perennial rhizome. The flowers and pseudo stem are used as a vegetable. The fibre is used for making ropes, mats, coarse paper and pulp. Peeled leaf sheaths are used as packing material. The juice of the stem dissolves kidney stones and acts as an antidote for snake biting. Unripe fruits and cooked flowers are good for diabetes (Azizah Mahmood et al., 2011; Mohammed Zafar Imam et al., 2011).

Butea monosperma (Lamk.) Taubert. (Fabaceae) is a medium sized tree with compound leaves. It is a valuable host tree for

Diospyros ebenum J.Konig. ex Retz. (Ebenaceae) an evergreen tree with a spreading crown. Fruits are edible. Tannin from raw fruits is used as preservative for wood.

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Ocimum sanctum L. (Lamiaceae) it is an aromatic shrub and environment purifier. The stem is made into beads and used as rosaries by Hindus. The leaf juice mixed with honey cures cough, cold, fever and bronchitis. Oil obtained from the leaf is an antiseptic and has an antimicrobia l and insecticidal property (Baskaran 2008; Dipak koche et al., 2011).

Achyranthes aspera L. (Amaranthaceae) a branched woody herb with spicate inflorescence. The plant is rich in potash and valued as green organic manure. The plant is pungent, purgative, diuretic and astringent. The flowering spikes ground into paste are used as an external application for poisonous insect bites (Niranjan Sutar et al., 2011; Sharma Raj Neeta et al., 2011).

Calotropis gigantea (L.) Dryand. (Apocynaceae, previously under Asclepiadaceae) is a large milky shrub and plant parts are covered with loose soft white wool. The latex is used in tannin industry to decolorizing, for removing hair and imparting yellow colour to hides. The floss obtained from the seeds is used for stuffing purposes (Chandrabhan Seniya et al., 2011; Himanshu Josi et al., 2011).

CONCLUSION

Imperata cylindrica (L.) Raeusch. (Poaceae) an erect perennial herb with a leafy culm. It is a good sand binder and protects the soil from erosion. The root of the grass is sweet, has a cooling effect and increases the flow of milk. Good fodder for cattle. The plant is used in Unani drug for treating snake bite. It is recommended in urinary disorders. Allmania nodiflora R. Br. Ex. Wight (Amaranthaceae) an erect straggling herb. Spikes yellowish flame coloured. The leaves are used as vegetable. It gives good organic manure to soil.

The temples of India are the hoary treasure not only for this country but also for the whole world. The present work ensures that our ancient ecological tradition and respect for the natural heritage is not lost in today`s fast changing life style and reminds the duty of every citizen to protect it. These Sacred Trees preserved through millennia by our ancestors as potential bio resources should be respected and conserved for the future generations. ACKNOWLEDGEMENTS The authors are grateful to UGC, New Delhi for financial assistance in the form of Major Research Project. Also thankful to the Head, Department of Botany and principal of Holy Cross College (Autonomous), Tiruchirappalli for the facilities provided and encouragement.

REFERENCES Amal Kumar Ghimeray., cheng – Wujin., Bimal Kumar Ghimire and Dong Ha Cho (2009). Antioxidant activity and quantitative estimation of azadirachtin and nimbin in Azadirachta indica A.Juss. grown in foot hills of Nepal. African journal of biotechnology. 8 (13): 3084–3091. Ambasta S P (1986). The useful plants of India. Publications & Information Directorate, CSIR, New Delhi. PP.16–405.

Ambersing Raj put., Suboth Chandra Pal and Bhagavan Patil (2011). Phytochemical

evaluation of leaves of Butea monosperma. International journal of pharmacy and pharmaceutical sciences. 3(3):189–191. Amirthalingam M (1998). Sacred trees of Tamil Nadu. The C.P. Ramaswamy Aiyar Foundation, Chennai.

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Mahmood., Nurziana Nagah and Muhammed Nor omar(2011). Phytochemical constituents and antioxidant activities in Musa paradisica flower. European journal of scientific research 66(2). 311–318.

Baskaran X (2008). Preliminary phytochemical studies and antibacterial activity of Ocimum sanctum L. Etanobotanical leaflets. 12:1236–1239. Bentley Robert and Trimen Henry (2000). Medicinal plants. Vol.I – IV. Periodic Expert Book Agency, New Delhi. Chakraberty C (1993). Ancient Hindu Medicine.1sted. Low price publications; Delhi. Chandrabhan Seniya., sumint singh Trivedia and Santiosh Kumar Verma(2011). Antibacterial efficacy and phytochemical analysis of organic solvent extracts of Calotropis gigantean, .journal of chemical pharmaceutical Research. 3(6): 330– 336. Dipak Koche., Syed Imran., Rupali Shirsat and Dyaneshwar Bhadange (2011). Comparative Phytochemical and Nutritional Studies of Leaves and Stem of Three Lamiaceae Members. Research Journal of Pharmaceutical, Biological and Chemical Sciences.2(3):1–4. Gamble J S (1967). Flora of the presidency of Madras. Vol.I – III Botanical Survey of India, Calcutta. Himanshu Joshi1., Gururaja M P. and Divya Suares ( 2011). Calotropis gigantea R. Br. (Asclepiadaceae): A Review. International Journal of Pharmaceutical Research. 3(1):10–14. Inder Kumar Makhija., Indra Prakash Sharma and Devang Khamar (2010). Phytochemistry and Pharmacological properties of Ficus religiosa: an

overview. Scholars Research Library.1 (4) : 171–180. Jain S K (2004). A manual of ethnobotany. Scientific Publishers, Jodhpur. Jeethu Anu Mathews., Karthikeyan M. and Annamalai A (2011). Antibacterial activity of Eugeniajambolana plant leaves extracts. An international journal of pharmaceutical sciences. 1728–1738. Khan L., Shane N., Yunis M. and Arfan M( 1999). An overview of plants used in the traditional system of medicine for the control of diabetes. Hamdard medicus .45(3):PP.38–45. Krishna Murti A., Upendra Kumar B. and Raghuvir Singh C( 2011). Antimicrobial activity and reversal of dexamethasone depressed healing by roots of Ficus racemosa L. Asian Journal of Traditional Medicines. 6 (5): 224–230. Mathew KM (1983). Flora of the Tamil Nadu C arnatic. The Rapinat Herbarium, St.Jose ph`s College, Tiruchirappalli. Minh Hien Ha A., Van Thi Nguyen A., Khac Quynh Cu Nguyen B, Emily LC Cheah C., Paul WS. and Heng C (2009). Antimicrobial activity of Calophyllum inophyllum crude extracts obtained by pressurized liquid extraction. Asian Journal of Traditional Medicines.4 (4): 141–146. Mohammad Zafar Imam and Saleha Akter( 2011). Musa paradisiaca L. and Musa sapientum L.:A Phytochemical and Pharmacological Review. Journal of Applied Pharmaceutical Science. 1(5):14–20. Niranjan Sutar., Ranju Garai ., Uma Shankar Sharma., Priyanka Goyal and Gangotri Yadav (2011). Pharmacognostic Studies Of The Achyranthes aspera Leaves.

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International Journal Of Comprehensive Pharmacy. 5 (10): 1–3. Poongothai1 A., Sreena K P., Sreejith K., Uthiralingam M. and Annapoorani S (2011). Preliminary phytochemicals screening of Ficus racemosa Linn. Bark. International Journal of Pharma and Bio Sciences.2(2): 431–434. Rajan S.,Gokila M., Jency P., Brindha P. and Sujatha R K (2011). Antioxidant and phytochemical properties of Aegle marmelos fruit pulp .International journal of current pharmaceutical research.3(2): 65–69. Rmanjaneyulu K., Bhargavi A., Raj Varma Buddaraj P., Pavani Siri K. and Meharvineela P (2011). Evaluation of phytochemical analysis and anti bacterial activity of Butea monosperma extracts. IJRAP.2(5): 1563–1565.

Shaw H R Airy and Willis J C (1973). A Dictionary of the flowering plants and Cambridge University ferns.8thed. Press, Cambridge. Sood S K., Thakur Vandana and Lakhanapal T N (2005). Sacred and magico- religious plants of India. Scientific Publication Jodhpur. Trivedi Priya Ranjan (1996). International environmental Laws. APHPublishing Corporation in Association with Indian Institute of Ecology and Environment, New Delhi. Uma B., Prabhakar K. and Rajendran S (2009). Invitro Antimicrobial Activity and Phytochemical of Ficus religiosa L. and Ficus bengalensis L. against Diarrhoeal Enterotoxigenic E. coli. Ethnobotanical Leaflets. 13: 472–74.

Shanbhag D A. and Amit Narayan Khandagale (2011). Application of HPTLC in the Standardization of a Homoeopathic Mother Tincture of Syzygium jambolanum. Journal of Chemical and Pharmaceutical Research. 3(1):395– 401.

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Sharma Raj Neeta., Bala Jyoti., Singh Anjuvan and Kaur Prabhjot (2011). Antibacterial potential of Achyranthes aspera Linn procured from Himachal Pradesh, Punjab and Haryana, India. Research journal of chemical sciences. 1(8): 80– 82.

Venkatesan D., Karrunakarn C M., Selva kumar S. and Palani Swamy P T (2009). Identification of phytochemical constituents of Aegle marmelos responsible for antimicrobial activity against selected pathogenic organism. Ethnobotanical Leaf lets .13: 1362–72.

Source of Support: Nil

Conflict of Interest: None Declared

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Review Article CERTAIN INDIAN MEDICINAL PLANTS SAFEGUARD AGAINST CANCER Nema Rajeev1*, Khare Sarita2, Jain Parul3, Pradhan Alka4 1,2,3,4

Sarojini Naidu Government Girls Post Graduate (Autonomous) College, Center for Microbiology & BioTechnology Research and Training, Shivaji Nagar, Bhopal â&#x20AC;&#x201C; 462016 (M.P.) * Corresponding Author: Email - rrsht.nema@gmail.com

Received: 01/04/2012; Revised: 29/05/2012; Accepted: 03/06/2012

ABSTRACT Herbal products are considered to be symbols of protection in comparison to the synthetic products that are regarded as risky to human life and environment. Some Medicinal plants have therapeutic potential due to the occurrence of natural antioxidants functioning as reducing agents, free radical scavengers and quenchers of singlet oxygen. Majority of their natural activity is due to bioactive compounds viz. phenol, polyphenolic, alkaloids, Saponin, and other secondary metabolites. And now the major challenge in pharmaceutical industry is drug improvement and discovery. This review discusses about some Indian medicinal plants with regard to their anti-cancerous property. Keywords: Anti-cancerous activity, Indian Medicinal plants, Brassica nigra, Carum carvi, Phyllanthus emblica, Trachyspermum ammi, Coriandrum sativum, Allium sativum, Spinacia oleracea

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INTRODUCTION Plant kingdom is a potential source of chemical constituents with antitumor and cytotoxic activities owing to their enormous propensity, which synthesizes a variety of structurally diverse bioactive compounds (Kim et al., 2005; Indap et al., 2006). The discovery of new drugs and their development into commercial products takes place across the broad scope of the pharmaceutical industry and research institutes. Attempts are underway to work out the therapeutic and anti-neoplastic properties of medicinal plants (Abo et al., 2000; Graf, 2000; Ankli, 2002; Neto, 2002). Consequently, herbal medicines have received much attention as substitute anticancer drugs. The rich and diverse plant sources of India are likely to provide effective anticancer agents. Medicinal plants can reduce or minimize the toxic side effects of chemotherapy and radiation treatment by reinforcing their cancer killing action. Some of the medicinal plants available in India which are anti-cancerous have been reviewed in this article. MATERIALS Brassica nigra L. (MUSTARD) Scientific classification - Kingdom: Plantae; (unranked): Angiosperms; (unranked): Eudicots (unranked): Rosids; Order: Brassicales; Family: Brassicaceae Juss. Genus: Brassica; species: B. nigra L. Brassica vegetables are highly regarded for their nutritional value and it is a good medicinal source for many diseases. In addition phytochemicals investigation indicate oleic acid, phenolics, carotenoids, selenium, glucosinolates and vitamin C present in barssicaceae mustard seed (Conaway et al., 2001; Fimognari et al., 2002) and offer broadspectrum support for caring against the ubiquitous cancer . Mustard leaves have been reported to possess many bioactive substances and antioxidant properties (Kim et al., 2003). Earlier reviews appeared dealing with biocidal, bio-herbicidal, antioxidant, anticancer activities of glucosinolates and their products

from Brassicaceae (Fahaey et al., 2001; Zukalova and Vasak 2002; Halkier and Gershenzon 2006; Vig et al., 2009). Coriandrum sativum L. (CORIANDER) Scientific classification - Kingdom: Plantae; (unranked): Angiosperms; (unranked): Eudicots (unranked): Asterids; Order: Apiales; Family: Apiaceae; Genus: Coriandrum; Species: C. sativum L.. C. sativum is well known for its antioxidant properties and some of its active components have been identified. Coriander contains active phenolic acid compounds, including caffeic and chlorogenic acid. The flavonoids include quercetin, keampferol, rhamnetin and apigenin. Most of these compounds are known to inhibit free radicals generated in the cellar system, when they are obtained through the diet. C sativum has been conventionally referred to as anti-diabetic, anti-inflammatory and cholesterol lowering (Chithra and Leelamma, 1999). Various activities of Coriandrum sativum include anthelmintic (Eguale T et al., 2007), anti-implantation (Al-Said MS et al., 1987 ), anti-microbial (Begnami AF et al., 2010), antimutagenic (Cortes-Eslava et al., 2004), antioxidant, hepato-protective (Samojlik I et al., 2010), anxiolytic, sedative and muscle relaxant (Emamghoreishi M, 2005). Trachyspermum (AJWAIN)

ammi

SPRAGUE.

Scientific classification: Kingdom: Plantae; (unranked): (unranked): Angiosperms; Eudicots; (unranked): Asterids; Order: Apiales; Family: Apiaceae; Genus: Trachyspermum; Species: T. ammi.Sprague. Trachyspermum ammi (L.) a well known spice is a traditional herb widely used for curing various diseases in both humans and animals. Aromatic chemicals present in Ajwain; inhibit other undesirable changes in food, affecting its nutritional quality, texture and flavor. Decoction of Ajwain seeds is used for treatments of abdominal discomfort, diarrhea, cough and stomach troubles (Anikumar et al., 2009). Fruit of Ajwain is

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reported to have antiseptic, antifungal, antibacterial and anthelminitic effects (Morsi, 2000). In T. ammi (Ajwain) a major phenolic compound, Thymol is present and has been reported to be an antispasmodic, germicide and antifungal agent (Nagalakshmi et al., 2000). The essential oil of T. ammi, the principle active constituents of the oil is phenols, mainly thymol (35 to 60%) and some carvacrol (Tsimidou and Boskou, 1994). Both the phenols Thymol and carvacrol are responsible for the antiseptic, anti-tussive and expectorant properties (Treas and Evans, 2002). Ajowan is an aromatic seed spices, generally used as a digestive stimulant or to treat liver disorders. Carum carvi L. (CARAWAY) Scientific classification - Kingdom: Plantae; (unranked): Angiosperms; (unranked): Eudicots; (unranked): Asterids; Order: Apiales; Family: Apiaceae; Genus: Carum; Species: C. carvi.L. The phytochemical essential oil of C. carvi collected from various countries has been widely studied (Samojlik, I et al., 2010).Many data indicated the essential oil is useful as an antimicrobial, antifungal, molluscidal, nematicidal, antioxidant and anti-aflatoxigenic activities, as well a potential drug used as a cancer preventing agent (Samojlik, I et al., 2010 and Zheng, G et al., 1992). In addition it is most commonly used in cases of gastric problems, flatulence and indigestion. It has been very efficient in relief of flatulent colic in infants. It is also known to boost the immune system and possesses antibacterial (Zheng, G. et al., 1992), antiulcerogenic (Khayyal MT et al., 2001), antitumor (Zheng G et al., 1992), anti-proliferative (Nakano Y et al., 1998) and anti-hyperglycemic effects.(Eddouks M et al., 2004 ). Spinacia oleracea L. (SPINACH) Scientific classification – Kingdom: Plantae; (unranked): Angiosperms; (unranked): Eudicots; (unranked): Core eudicots; Order: Family: Amaranthaceae; Caryophyllales; Genus: Spinacia; Species: S. oleracea. L.

Spinacia oleracea L. is commonly reported to be a good source of minerals, vitamin B complex, ascorbic acid, carotenoids (bcarotene, lutein, zeaxanthine), flavonoids, apocyanin and p-coumaric acid (Bergan et al., 2001). The entire plant is used as a remedy for urinary calculi and the leaves are used for bowel and lung inflammation (Jain and De Fillipps, 1991). Apart from having nutritional value, it has been also credited with various biological activities like virus inhibitor (Adam G et al., 2008), anthelmentic (Patil UK et al., 2009), antioxidant, (Verma RK et al., 2003) hepatoprotective (Gupta R S et al., 2006)and reducing risk of breast cancer (Longnecker MP et al., 1997). Extracts of spinach leaves show high anti-oxidative activities and are well tolerated in animal studies (Lomnitski L, et al., 2003). Allium sativum L. (GARLIC) Scientific classification – Kingdom: Plantae; clade: Angiosperms; clade: Monocots; Order: Asparagales; Family: Amaryllidaceae; Subfamily: Allioideae; Genus: Allium; Species: A. sativum L. The effects of garlic on health with its possible preventive effects on the development of cancer in humans have been mentioned in previous reviews. In addition A. sativum has free radical scavenging activity, immune system modulation and direct cytotoxic effect on cancer cells (Abdullah et al., 1988). Now a day’s numerous epidemiological, clinical and laboratory studies have demonstrated the role of garlic in cancer prevention ( Flieischauer et al., 2001, Galeone et al., 2006.) especially in relation to digestive tract cancers, including esophageal and stomach cancers (Berspalov et al., 2004). There is also promising research evaluating the use of garlic in leukemic melanoma (Taylor et al., 2006) and neuroblastoma (Karmakar et al., 2007) cell lines.

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Phyllanthus emblica L. (AMLA OR INDIAN GOOSEBERRY)

cell lines at doses of 50–100 µg/ml. (Ngamkitidechakul et al., 2010).

Scientific classification – Kingdom: Plantae; Division: Flowering plant; Class: Magnoliopsida; Order: Malpighiales; Family: Phyllanthaceae; nTribe: Phyllantheae; Subtribe: Flueggeinae; Genus: Phyllanthus; Species: P. emblica L.

CONCLUSION

Preliminary research on Phyllanthus emblica in vitro demonstrates antiviral and antimicrobial properties (Saeed S et al., 2007). There is also evidence that its extracts induce apoptosis and modify gene expression(Penolazzi L et al., 2008). In addition Phyllanthus emblica leaves, bark or fruits have shown potential efficacy against laboratory models of disease, such as for inflammation, cancer, age-related renal diseases, and diabetes (Yokozawa T et al., 2007), ( Rao TP et al., 2005). The potential anticancer effects of aqueous fruit extract of P. emblica was tested in several different human cancer cell lines such as A549 (lung), HepG2 (liver), HeLa (cervical), MDA‐MB‐231 (breast), SKOV3 (ovarian) and SW620 (Colorectal). P. emblica extract significantly inhibited the growth of several human cancer

Although a large number of synthetic drugs are being added to the world of modern pharmacopoeia, but still no system of medicine in the world could solve all the health problems. Several diseases like AIDS, Birdflue, and Cancer etc., therefore the search for new therapeutic constituents from plants is genuine and urgent. In India, there is an ocean of knowledge about medicinal plants and rich medicinal flora, but still only a few pearls have been searched as therapeutic agents. There are large numbers of indigenous plants left which have not been investigated thoroughly from modern scientific view or their curative values have not been recognized. Thus there is an urgent need for systematic phytochemical investigation of those plants which have not been investigated systematically. ACKNOWLEDGEMENT The authors express gratitude Prof. Dr. Usha nair Department of chemistry government, M.V.M. College for her kind support.

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Zhao H, Lin J, Grossman HB, Hernadez L. M, Dinney CP and Wu X. (2007). Dietary isothiocyanates GSTM 1, GSTT 1, NAT 2 polymorphism and bladder cancer risk. International Journal of Cancer 120:2208–2213. Zheng G, Kenney PM, Lam LKT. (1992) Anethofuran, carvone, and limonene:

Source of Support: Nil

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Conflict of Interest: None Declared

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Short communication ETHNO BOTANICAL SURVEY OF MEDICINAL PLANTS OF SARGODHA REGION AND ITS VICINITIES Ghani Abdul1*, Hasan Naqi2, Ishtiaq M3 1, 2, 3 *

Department of Biological Sciences,University of Sargodha, Sargodha. Pakistan

Corresponding Author: ghaniuaf@yahoo.com; Phone: 0307-6746099 Received: 20/04/2012; Revised: 27/05/2012; Accepted: 31/05/2012

ABSTRACT In recent times, focus on plant research has increased all over the world and a large body of evidence has collected to show massive potential of medicinal plants used in various traditional systems. Medicinal plants based traditional systems of medicines are playing a vital role in providing health care to large sections of population, especially in developing countries. Interest in them and utilization of herbal products produced based on them is increasing in developed countries also. To gain maximum benefits it is necessary to have minimum basic level information on their different aspects. This article has been based on the results of an ethno-botanical research conducted in Sargodha region. The main objective of this study was to enlist the wealth of medicinal plants. In total 23 species, belonging to 17 families of wild herbs, shrubs and trees, were found to be used as medicinal plants by the inhabitants in the Sargodha region.

Keywords: Sargodha region, identification and uses of medicinal plants

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INTRODUCTION Medicinal plants are consumed worldwide for the treatment of several diseases and plant species form an important source of raw material for pharmaceutical industries. In recent decades the use of herbal medicines has increased worldwide. This may be because of the fact that the side-effects of these materials are often lower than synthetic drugs apart from the higher costs of many conventional pharmaceutical formulations (Rates, 2000). Therapeutic plants have always been valued as a mode of treatment of variety of ailments in folk cultures and have played a very important role in discovering the modern day medicines with novel chemical constituents (Chan, 2003; Haider et al., 2004; Devi et al., 2008; Shirin et al., 2010). Although the efficacy of medicinal plants for curative purposes is often accounted for in terms of their organic constituents like essential oils, vitamins, glycosides etc. Medicinal plants come into preparation of various drugs singly or in combination or even are used as the principal source of raw material for the other medicines (Mohanta et al., 2003). Sargodha is a district of Punjab province, Pakistan. It is an agricultural district. Wheat, rice, and sugarcane are its main crops. The Sargodha district and region are also famous for citrus fruit. The district has an area of 5,864 km2. The term Sargodha has its origins in the words Sar, meaning pond (Talab), and Godha, meaning sadhu (Baba). Just over a century ago, there was a well owned by a Hindu named Goodha, the well was known as "Gool Khuuh" (round well), and travelers used it as a resting place. There is a market at the place where Gool Khuuh used to exist and a grand mosque above the market and it is known as "Goal Chowk", which is the center of downtown Sargodha today. Sargodha mainly

comprises of flat, fertile plains, although here are a few small hills on the SargodhaFaisalabad Road. The River Jhelum flows on the western and northern sides, and the River Chenab lies on the eastern side of the city. The city has a climate of extreme heat and cold. MATERIALS AND METHODS This research was conducted during summer 2001–2002 in Sargodha valley. The plants were collected, dried and preserved for identification. They were identified with the help of available literature [Stewart 1972, Nasir and Ali 1971–95]. The information about the medicinal uses of the plants was obtained from the local people through questionnaires. The outcome of the results were rechecked and compared with literature like that of Rubina [1998], Ali and Fefevre [1996] and Khalid [1995]. The data was analyzed and indigenous knowledge was documented. RESULT AND DISCUSSION The importance of the herbal medicine can be well understood by the saying of Hippocrates “Let medicine be your food and food your medicine”. The botanical sources of large number of folk medicines, found therapeutically effective in indigenous systems, are still unknown (Ahmad et al., 2008, 2009; Abbasi et al., 2009; Shah et al., 2009). The research revealed that local folk utilizes 41 species of plants belonging to 29 families for various purposes (Table 1). The people of the region are generally ignorant about the medicinal and economic importance of these plants. Out of the 41 medicinal plants, only 20 species were known to the locals and the rest of the species of high medicinal and economic values were completely unknown to the local community of the area.

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TABLE 1: CHECK LIST OF MEDICINAL FLORA S.No

Botanical Name

Vernacular Name

Family

Part Used

Ethno medicinal Uses

1.

Plantago ovata Peganum harmala Boerhavia diffusa Spinacea oleracea Solanum nigrum

Ispaghul

Plantaginaceae

Hermal

Zygophyllaceae

It sit

Nyctaginaceae

Palak

Chenopodiaceae

Makoo

Solanaceae

Seeds fruit and leaves Whole Plant Leaves, Bark Leaves and stem Whole Plant

Tribulus terrestris Cynodon dactylon Agaricus campestris L.

Bhakra

Zygophyllaceae

Khuble ghas

Poaceae

Whole Plant Roots

Antidiarrhoeal, Constipation and laxative. Brain tonic, insecticidal and Viral diseases. Antiamoebic, Antibacterial, pain relief, improves and protects eyesight. Anemia, Bone's Tonic and produce fresh blood. Antiseptic, tonic in fever, dyspepsia, scabies, skin infection, dental problem. Painful urination

Khunbii Mushroom

Agaricaceae

Whole part

Alliaceae

Bulb, leaves and seeds.

Alliaceae

Bulb

2. 3. 4. 5.

6. 7. 8.

9.

Allium L.

cepa Piaz

10.

Allium sativum L.

11.

Beta vulgaris Choqandar L

Chenopodiaceae

Roots and leaves

12.

Citrulus lanatus

Tarbooz

Cucurbitaceae

Fruit and seeds.

13.

Hordeum vulgare L

Jao

Poaceae

Fruit

14.

Lagenaria siceraria

Kaddo

Cucurbitaceae

Fruit and leaves

15.

Lawsonia inermis

Mehndi

Lythraceae

Leaves, branches and flowers

16.

Citrus aurantifolia

Laymun

Rutaceae

Fruit, juice, bark and seeds

Lahson

Diuretic and Laxative Eye diseases, germicide, arthritis, paralysis, parkensis, muscle pain, headache and dizziness. Stomach diseases, cholera, Diarrhea, throat infection, common cold cough, fever, influenza, ear pain, appetizer, headache, hepatitis, piles, eye diseases. Wound healer, paralysis, digestive problems asthma, intestinal pain worms, cough, headache, tuberculosis. Liver infection, muscle weakness, skin disease, hepatitis, kidney pain, headache. Clears the urine and stomach, facial beauty, increase immunity, jaundice and have cold effects. Fever, weakness, increase immunity, heart diseases, kidney pain, intestinal ulcer maintain cholesterol level, jaundice and have cooling effect. Headache, fever, Madness, Piles, lungs infection, common cold, kidney and liver disorder and heart diseases. Wound healer, headache, increase memory, cool effect, protect the rupture of skin, skin softer; heal the muscle and chicken pox. Antidote, remove spot from skin, piles, stomach disorder, hepatitis, heart diseases, intestinal pain, pimples.

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Lens culinaris

Masoor

Papilionaceae

18.

Nigella sativa

Clongy

Ranunculaceae

19.

Phoenix dactylifera

Khajoor

Arecaceae

20.

Trigonella foenumgraecum L.

Maithi

Papilionaceae

21.

Zingiber officinale Roscoe.

Adrak or Sonth

Zingiberaceae

22.

Cucumis Sativus

Cucumber

Liliaceae

23.

Zizyphus jujuba

beri

Rhamnaceae

CONCLUSION The area needs proper protection for the conservation and survival of bio-resources with the help of local people. Regular chemical screening of medicinal plants and their useful parts collected from the fields in different seasons should be done. Small scale processing units and regional repositories for the valued

Seeds

Maleness, measles, paralysis, common cold, parkensis, face clearness, eye infection, digestive diseases. Seeds Hysteria, common cold, asthma, constipation, urine blockage, gastric acute, gastro enteritis, diabetes, kidney stone, baldness, gas trouble, brain disorders, pneumonia, cough. Fruit Heart diseases, skin diseases, antidote, swelling of kidney, intestinal pain, heart attack, wound healer, diarrhea, stomach pain, piles, liver disorders. Seed and Throat infection, swelling, body pain, leaves. cough, stomach pain, piles, dandruff baldness, lungs infection, diabetes, ulcer, diarrhea and gas trouble, back pain, seeds in powder form used for diabetes. Fruit Intestinal pain, anorexia, dyspepsia, headache, diarrhea, constipation, intestinal swelling, digestive stimulant, cooling effect on body. fruit. Sedative and diuretic, anti-rheumatic and anti-inflammation of the eyes. Stimulant appetite. Fruit, Jaundice, Skin disease, Carminative, leaves and Sedative. roots

Drugs should be established. To prevent the extinction of these medicinal species, ex-situ conservation efforts may be undertaken to grow the sensitive species by acclimatizing them and in situ conservation of as many species possible could be done with the help of voluntary organizations and local NGO’s. If done this could be an asset for the future generations to come.

REFERENCES Abbasi AM, Khan MA, Ahmad M, Zafar M, Khan H, Muhammad N, Sultana S (2009). Medicinal plants used for the treatment of jaundice and hepatitis based on socio-economic documentation. Afr. J. Biotechnol., 8(8): 1643–1650.

Ahmad M, Khan MA, Hasan A, Zafar M, Sultana S (2008). Chemotaxonomic standardization of herbal drugs Milk thistle and Globe thistle. Asian. J. Chem., 6(20): 4443–4459. Ahmad M, Khan MA, Zafar M, Hasan A, Sultana S, Shah GM, Tareen RB (2009). Chemotaxonomic

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authentication of Herbal Drug Chamomile. Asian J. Chem., 21(5): 3395–3410. Ali, S.I. and Y.J. Nasir.1990–92. Flora of Pakistan. No. 191–193. Department of Botany, University of Karachi and National Herbarium, PARC, Islamabad. Chan

K (2003). Some aspects of toxic contamination in herbal medicines, Chemosphere. 52: 1361–1371.

Devi NK, Sharma NH, Kumar S (2008). Estimation of essential and trace elements in some medicinal plants by PIXE and PIGE techniques, Nucl. Instrum. Meth. Phys. Res. B 266: 1605– 1610. Haider S, Naithani V, Barthwal J, Kakkar P (2004). Heavy metal content in therapeutically important medicinal plants, Bull. Environ. Contam. Toxicol. 72: 119–127. Mohanta B, Chakraborty A, Sudarshan M, Dutta RK, Baruah M (2003). Elemental

Source of support: Nil

profile in some common medicinal plants of India. Its correlation with traditional therapeutic usage. J. Radioanalytical, Nuclear. Chem., 258(1): 175–179. Nasir, Y.J and Rafiq.R.A (1995). Wild Flowers of Pakistan. Edited by Roberts T. J., Oxford, University Press, Karachi: pp 298. Rates, SMK. (2000). Review: Plants as a source of drugs. Toxicon. 39:603–613. Shah GM, Khan MA, Ahmad M, Zafa M, Khan AA (2009). Observations on antifertility and abortifacient herbal drugs. Afr. J. Biotechnol. Shirin K, Imad S, Shafiq S, Fatima K (2010). Determination of major and trace elements in the indigenous medicinal plant Withania somnifera and their possible correlation with therapeutic activity. J. Saudi Chem. Soc. 14: 97– 100.

Conflict of Interest: None declared

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Review Article EFFICACY OF GLYCYRRHIZA GLABRA LINN. (YASTIMADHU) IN LEARNING, MEMORY AND COGNITIVE ACTIVITY –CURRENT FINDINGS AND FUTURE AVENUES Sheshagiri Srihari1*, Patel Kalpana Shanthibhai2, Shrikrishna Rajagopala3 1, 2, 3

Department of Kaumarabhritya, IPGT & RA, Gujarat Ayurved University, Jamnagar, Gujarat, India – 361 008 *Corresponding Author: E-mail: dr.srihari@rediffmail.com Mob. +919722188631

Received: 08/05/2012; Revised: 31/05/2012; Accepted: 04/06/2012

ABSTRACT Glycyrrhiza glabra Linn, a novel drug with several well known medicinal properties, has been reviewed here for current findings with regards to its efficacy in improving learning, memory and cognitive activity. A methodical review has been conducted on pharmacological, clinical study articles and dissertation works published from 2004 to July 2008, using PubMed and manual search of bibliographies as source. The pharmacological researches indicate that Glycyrrhiza glabra Linn shows improvement in memory, learning and spatial memory retention enhancement effects in vivo via multiple mechanisms of action, a clinical study on the synthetic derivative of glycyrrhetinic acid shows promising result in improving cognitive functions, enhancement of Medha (Intellect and Memory) was also observed in 2 dissertation works based on the properties of the drug. The present review is on some of the relevant studies on Glycyrrhiza glabra Linn wherein the methods employed, results obtained and mechanism of action of the drug have been explained. The review also emphasizes on the future thrust areas of research upon this drug on the same back ground.

Keywords: Glycyrrhiza glabra Linn, Learning, Memory, Cognitive activity, Medha

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www.gjrmi.com INTRODUCTION

MATERIALS

Plenty of plants in traditional medicine are being used in age-related brain disorders for improvement of memory and cognitive function. [1-3] Glycyrrhiza glabra Linn is one among them and is known as “Yastimadhu” in the traditional Indian system of medicine “Ayurveda”. This traditionally much valued plant is one among the four herbs mentioned in ‘Charaka Samhitha’ – An Ancient Indian Medical treatise, as “Medhya Rasayana” (drug promoting intellect and memory) [4].

Glycyrrhiza glabra Linn of Fabaceae Photo slides 1 & 2) is a perennial plant, family (P grows up to more than 1 m in height, erect, with highly developed stoloniferous roots. Leaves compound, 9–17 alternate imparipinnate leaflets, oblong to ellipticallanceolate, acute or obtuse; racemes loose, shorter than the leaves or a little longer. Flowers 1 cm long, flat pods oblong to linear, 1–3 cm long by 6mm wide, more or less densely echinate glandular, many seeded or abbreviated 2- or 3-seeded. [5, 6] The commercial name of the dried rhizome and root of the plant is liquorice. [7]

Even though Yastimadhu has been praised as an effective Medhya drug and mentioned as second best Medhya rasayana, [4] very minimal works have been carried out in the field of Ayurvedic research owing to the above activity and lots are to be still done. With an aim to reestablish and interpret the Medhya property of Yastimadhu, a methodical collection of works done pertaining to the above said properties on pharmacological and clinical background has been done here.

Roots and Rhizome are the officinal part, which are nearly cylindrical pieces, up to 1m long and 5–20 mm in diameter; externally, the bark is brownish grey to dark brown, longitudinally wrinkled, occasionally bearing small dark buds in rhizomes or small circular or transverse rootlet-scars in roots. The peeled root is yellow, smooth, fibrous, finely striated; fracture fibrous in the bark and splintery in the wood; internally, bright yellow. A distinct cambium ring separates the yellowish grey bark from the finely radiate yellow wood; central pith, only in rhizomes. [5, 8, 9]

Photo slides 1 & 2: Plant and roots of Glycyrrhiza glabra Linn

Source of Photographs: 1. http://www.hauenstein-rafz.ch/de/pflanzenwelt/pflanzenportrait/stauden/Suessholz-Glycyrrhiza-glabra.php 2. http://parthpratisthan.org/plants/common_medicinal_plants-_7

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The main compounds accountable to bring about the pharmacological effects of G. glabra are Saponins, Flavonoids, Alkaloids, Tannins, Steroids, etc. The major constituents are triterpene saponins. Glycyrrhizin is generally regarded as the active principle of G. glabra and is responsible for its sweetness, which is 50 times more than that of sucrose. [10] Glycyrrhizin (glycyrrhizic acid, glycyrrhizinic Properties: 16 Rasa (Taste) Guna (Quality) Virya (Potency) Vipaka Karma (Action)

: : : : :

Prabhava (Special Action)

:

acid) is the major component (2–9%); minor components occur in proportions that vary depending on the species and geographical location. [11-13] Glycyrrhizin occurs as a mixture of potassium and calcium salts. [14] It is a monodesmoside, which on hydrolysis releases two molecules of D-glucuronic acid and the aglycone glycyrrhetic (glycyrrhetinic) acid (enoxolone). [15]

Madhura (Sweet) Guru (Heavy), Snigdha (oily) Sheetha (Cold) Madhura (Sweet) Balya (Immune booster), Cakshushya (Vision promoter), Varnya (Complexion promoter), Vatapittajit (Reduces vitiated Vata and pitta humors), Raktaprasadana (enhancing the qualities of blood) Medhya (drug promoting intellect and memory)

Pharmacokinetics: Glycyrrhizic acid, the main constituent of licorice after oral administration, is hydrolyzed to glycyrrhetic acid by intestinal bacteria possessing a specialized ßglucuronidase. [9, 17] After oral administration, glycyrrhetic acid is rapidly absorbed and transported via carrier molecules to the liver. In the liver it is metabolized to glucuronide and sulfate conjugates, which are subsequently rehydrolyzed to glycyrrhetic acid. Glycyrrhetic acid is then reabsorbed, resulting in a significant delay in terminal clearance from plasma. [18] After oral administration of 100 mg glycyrrhizin (Fig 1.) in healthy volunteers, no glycyrrhizin was found in the plasma but glycyrrhetic acid was found at < 200 ng/ml. In the 24-hour period after oral administration, glycyrrhizin was found in the urine, suggesting that it is partly absorbed as an intact molecule. [15]

Dosage: Unless and otherwise prescribed, average daily dose of crude plant material is 5– 15 g, corresponding to 200–800 mg of glycyrrhizin and should not be used for longer than 4–6 weeks without medical advice. [19] Doses of other preparations should be calculated accordingly. [19]

Adverse reactions: No adverse reactions have been associated with the drug when used within the recommended dosage and treatment period. Prolonged use (6 weeks) of excessive doses (>50 g/day) can lead to pseudoaldosteronism, which includes potassium depletion, sodium retention, oedema, hypertension, and weight gain. [14, 20, 21] In rare cases, myoglobinuria and myopathy can occur. [22] Toxicity: The acute toxicities of licorice extract and glycyrrhizin salts are low with oral LD50s generally greater than 4 gm glycyrrhizinate/kg b.w in mice and rats. Other in vivo studies reported no teratogenic effects when glycyrrhizin salts were administered maternally to mice, rats, hamsters, or rabbits during gestation at doses as high as 1000 mg/kg/day. However, dominant lethal testing in male rats suggests that an intake of 4000–5000 mg glycyrrhizin/kg/day could lead to mutagenic effects in offspring. Microbial tests found that licorice extract and glycyrrhizinates were nongenotoxic and had some anti-genotoxic properties. There is abundant evidence from both case reports and clinical studies that the habitual consumption of glycyrrhizin results in adverse effects marked by the development of pseudohypercorticosteroidism. [23]

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Current Pharmacological findings on Learning, Memory: (1) Memory strengthening activity of Glycyrrhiza glabra Linn in exteroceptive and interoceptive behavioral models [24]:Aqueous extract of G. glabra was administered for 7 successive days in separate groups of mice in three doses (75, 150, and 300 mg/kg p.o.). An exteroceptive and interoceptive behavioral models of memory was used. The dose of 150mg/kg of the aqueous extract of liquorice significantly improved learning and memory of mice. Furthermore, this dose reversed the amnesia induced by diazepam (1 mg/kg i.p.), scopolamine (0.4 mg/kg i.p.), and ethanol (1 g/kg i.p.). Anti-inflammatory and antioxidant properties of liquorice may be contributing favorably to the memory enhancement effect. Since scopolamineinduced amnesia was reversed by liquorice, it is possible that the beneficial effect on learning and memory may be because of facilitation of cholinergic transmission in brain. (2) Memory enhancing activity of Glycyrrhiza glabra Linn in mice [25]:- With the same methodology as the above, ‘Elevated plus-maze’ and ‘passive avoidance paradigm’ were employed to test learning and memory. The dose of 150 mg/kg of the aqueous extract of liquorice significantly improved learning and memory of mice. (3) The central cholinergic pathways play a prominent role in the learning and memory processes. Acetyl cholinesterase is an enzyme that inactivates acetylcholine. In a study undertaken to evaluate the “Comparative brain cholinesterase-inhibiting activity of Glycyrrhiza glabra Linn, Myristica fragrans, Ascorbic acid, and Metrifonate in mice,” [26] Aqueous extract of G. glabra (150 mg/kg p.o. for 7 successive days), nhexane extract of Myristica fragrans seeds (5mg/kg p.o. for 3 successive days), ascorbic acid (60 mg/kg i.p. for 3 successive days), and metrifonate (50 mg/kg i.p.) were

administered to young male Swiss albino mice. Acetyl cholinesterase enzyme was estimated in brains of mice. Glycyrrhiza glabra, Myristica fragrans, Ascorbic acid, and Metrifonate significantly decreased acetyl cholinesterase activity as compared to their respective vehicle-treated control groups. (4) Spatial memory retention in rats [27]:Aqueous extract (GE) of G. glabra (Fabaceae) was administered systemically to rats and the time course of the effects on the spatial memory retention in the Morris water maze was studied. The dose of glycyrrhizin (GL), i. e., 0.5, 2.5 and 5 mg/ml in daily water intake of GE was administered to three groups of rats. The first, second and third groups received GE for 1, 2 and 4 weeks, respectively (each group included 3 subgroups). After terminating the treatments, all animals were trained for four days; each day included one block and each block contained four trials. Test trials were conducted 48 h after the completion of the training period. GE treatment decreased both escape latency and travelled distance, suggesting significant spatial memory retention enhancement by GE. (5) Effect of glabridin on learning and memory in mice [28]:- Glabridin (Fig 2) was isolated from the roots of G. glabra and its effects on cognitive functions and cholinesterase activity were investigated in mice. Glabridin (1, 2 and 4 mg kg (-1), P. O.) and piracetam (400 mg kg (-1), i. p.), a clinically used nootropic agent, were administered daily for 3 successive days to different groups of mice. The higher doses (2 and 4 mg kg (-1), P. O.) of glabridin and piracetam significantly antagonized the amnesia induced by scopolamine (0.5 mg kg (-1), I. P.) in elevated plus maze test and passive avoidance test. Furthermore, both glabridin (2 and 4 mg kg (-1), P. O.) and metrifonate (50 mg kg (-1), I. P. used as a standard drug), both remarkably reduced the brain cholinesterase activity in mice compared to the control group.

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GJRMI, Volume 1, Issue 6, June 2012, 24 247–257 Fig 1: Glycyrrhizin

Fig 2: Glabridin

Clinical studies on cognitive activity of Glycyrrhiza glabra Linn: (1) Two randomized, double double-blind, placebo-controlled crossover studies [29] were conducted on Carbenoxolone (Fig 3) a synthetic derivative of glycyrrhetinic acid upon on healthy volunteers and type 2 diabetic. 11β-HSD HSD inhibitor carbenoxolone was administerd 100 mg, orally, three times per day for 4 weeks with placebo in ten healthy volunteers. During each phase, subjects also received amiloride (10 mg/day) to prevent renal mineralocorticoid excess excess.[30] The two phases were separated by an 8-week week washout period. lve subjects with stable type 2 Twelve diabetes were enrolled. In this study the subjects were administered with 11β-HSD inhibitor carbenoxolone 100 mg mg, orally, three times per day for 6 weeks with placebo. During each phase, subjects also received amiloride (10 mg/day) to prevent renal

Fig 3: Carbenoxolon

mineralocorticoid excess. The two phases were separated by a 12--week washout period. Cognitive ability was assessed with the help of various tools in a similar manner for both healthy volunteers and type 2 diabetes subjects.. Nonverbal reasoning was evaluated evalu with Raven's Standard Progressive Matrices by using the number correct in 20 min.[31] Verbal fluency, thought to assess executive function and semantic memory, was assessed with the Controlled Word Association Test. Test [32] Verbal memory was evaluated with a test of list-learning, learning, the Rey Auditory-Verbal Auditory [32] Learning Test (AVLT) and paragraph recall, with the Logical Memory (immediate and 30-min-delayed) delayed) subtest of the Wechsler Memory Scale-Revised Revised. [33] Visuo-spatial memory y was evaluated with the Visual Reproduction (immediate and 30-min30 delayed) subtest of the Wechsler Memory Scale-Revised. [33] Attention and processing speed were evaluated with the Digit-Symbol Digit Substitution Test from the Wechsler Adult

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Intelligence Scale-Revised. [34] Prior general cognitive ability was assessed with the National Adult Reading Test. [35] Mood was assessed with the University of Wales Institute of Science and Technology (UWIST)-Mood Adjective Checklist [36] (MACL) and the Hospital Anxiety and Depression Scale (HADS). [37] The result showed improved verbal fluency (P < 0.01) after 4 weeks in 10 healthy elderly men (aged 55-75 year) and improved verbal memory (P < 0.01) after 6 weeks in 12 patients with type 2 diabetes (52–70 year). (2) In a dissertation work, “A study on the effect of Medhya Rasayana in school going children,” [38] 11 school going children were administered with one tablet of 500 mg containing equal quantity of Guduchi (Tinospora cordifolia), Yastimadhu (G. glabra), Shankhapushpi (Convolvulus pluricaulis) and Mandukparni (Hydrocotyle asiatica) twice daily with milk for a period of two months. The above drugs showed highly significant (P < 0.001) increase in Medha (Intellect) by 28.29%. The effect on Smriti (Memory) in treated group at the end of treatment was highly significant (P < 0.001) by 30.36%. The Seguin Form Board test battery was used for assessment which gave a specific reading associated with Visuo-motor and Visuo-perceptual areas. The mode of action of these compounds was explained based on the pharmacodynamics of the drugs present in the compound. The ushna virya drug Guduchi could be helpful mainly in Grahana (Grasping power) functions. The sheetha virya drugs Yastimadhu, Shankhapushpi and Mandukparni could be helpful mainly in Dharana (Retention power) functions. (3) In a dissertation work, “The Conceptual and Applied Study of Medha and Manas in Ayurveda,” [39] a placebo controlled clinical study; Volunteers were selected between the age of 16–25 years irrespective of their sex and religion. In Group A, Yastimadhu (G. glabra) granules were administered in the dose of 4 gm twice a day with milk as Anupana (Adjuvant with

medication) for 30 days duration. In Group B, Granules of roasted Suji were administerd at the dose of 4 gm twice a day with milk as Anupana and for 30 days duration+ Sattvavajaya Cikitsa (Psychological and Behavioural therapy). P.G.I. Memory scale of National Psychological Corporation, Agra had been adopted for the assessment of Medha before and after treatment. In group A, 17 volunteers completed the treatment while in group B, 15 volunteers completed the treatment. Group A showed a statistically highly significant improvement in P.G.I. memory scale and long term memory recall. Again the mode of action has been attributed to the pharmacodynamics of the drug Yastimadhu. DISCUSSION ‘Ayurveda’ the science of life has in store a vast knowledge of medicinal plants, metals and minerals, the medicinal efficacy of which are probably assessed through immense clinical trials and errors without any aide of modern equipments. It is amazing to the best of our knowledge, after exploring various pharmacological and clinical studies done on this novel drug Yastimadhu that these findings were in point of fact indicated thousands of years back in various treatises of Ayurveda. The current findings strongly uphold the rich knowledge of traditional systems of medicine and give a scientific revalidation of mode of action of the drug which is indeed the need of the hour. Current findings advocated that the aqueous extract of G. glabra at 150 mg/kg for 7 days can significantly improve learning and memory of mice as noted on ‘elevated plus maze’ and ‘passive avoidance paradigm’ tests. The findings also suggested that this dose reversed amnesia induced by diazepam, scopolamine and ethanol in mice. The same result was also observed after administration of glabridin (aqueous extract of G. glabra) an active isoflavone which remarkably reduced the brain cholinesterase activity (which plays a prominent role in learning and memory) in mice at 1, 2, and 4 mg/kg.[25] Aqueous extract

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of G. glabra and glycyrrhizin showed significant results on spatial memory retention of rats in Morris water maze. Improv Improvement in verbal fluency in healthy adults and verbal memory in type 2 diabetics was seen after administration of carbenoxolone (a synthetic derivative of glycyrrhetinic acid acid). The study suggested that 11β-HSD1 inhibition may be a new approach to prevent/ameliorate eliorate cognitive decline. By analysing the above mentioned few clinical studies it can be understood that G. glabra and its contents is definitely having a significant potential in enhancing learning and memory via multiple mode of action. Medha can be understood by Grahana Shakthi (Grasping power) [40] for which proper functioning of smriti and buddhi is a vital essentialilty. The above pharmacological studies give us a clear reflection about the ability of Yastimadhu to influence smriti and medha shakthi. The clinical studies re re-ascertain this action of the drug in healthy and diseased human volunteers too. Memory) has been Medha (Intellect and Memory said as the function of Pitta.. Balanced state of Pitta is responsible for proper functioning of Medha. The dominant rasa (Taste) of

Fig 4: Liquiritigenin

Yastimadhu is Madhura (Sweet). Madhura rasa is said to be ‘Sadindriya Sadindriya prasadaniya’ prasadaniya (Nourishing five sense organs and ManasMind)) and therefore has a direct effect over the site of these Indriyas (Sense organs) i.e. Sirah (Head). Madhura rasa, rasa Sita Virya and Madhura Vipaka of Yastimadhu pacifies Pitta. By Guru, Snigdha Guna and Madhura Vipaka it controls the Chala Guna of Vata. Vata is the controller and stimulator of Mind. As Medha is closely related to Manas the factors affecting Manas will affect Medha too. Various drugs and combinations promoting Medha and Smriti have been mentioned in Ayurveda. But choosing an appropriate drug for the necessity is left to Yukthi (logic) of the physician. On observing the results and mode of actions of G. glabra in pharmacological studies, we can interpret that it can be a drug of choice in patients suffering from Smriti hrasa (reduced memory) and Smriti nasha (dementia). The drug also shows its therapeutic efficacy in improving the Medha of school going children,, suggesting that it could be an effectivee medicine in children suffering from learning disabilities and scholastic backwardness.

Fig 5: Isoliquiritigenin

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Test

In vivo

Oral

GJRMI, Volume 1, Issue 6, June 2012, 247–257

Table 1: Memory-improving effects of Glycyrrhiza glabra Linn 29, 47 Test materials Endpoint/ Test model Mechanisms Effects /dose biomarkers

Extracts 75–300 mg/kg, 7d diet 0.5 or 1%, 6w

Glabridin 1–4mg/kg, 3d Carbenoxolone (Synthetic derivative of glycyrrhetinic acid) Medhya rasayana Tablets Yastimadhu Granules

Reference

Diazepam treated mice

Elevated plusmaze test

Scopolamine treated mice

passive avoidance test Morris watermaze test TBARS/ Catalase/AChE

Quench oxidative stress Inhibit AChE

ChE

Inhibit ChE

Various cognitive tests

11β-HSD inhibition

Improves cognitive functions

[27]

Seguin Form Board test battery

Pharmacodynamics of the drug

Improves Medha

[38]

P.G.I Memory scale and long term memory recall

Pharmacodynamics of the drug

Improves Medha

[39]

Aβ 25–35 treated mice Innate Mice Healthy volunteers & Type2 Diabetes patients Healthy volunteers (School going childrens) Healthy volunteers

[23] Cholinergic [22] Improves memory

[40]

[24]

#Acetylcholinesterase (AChE); cholinesterase (ChE); thiobarbituric acid-reactive substances (TBARS); β amyloid (Aβ); 11β-Hydroxysteroid dehydrogenase (11β-HSD).

Future avenues Further studies are desirable to explore the role of drug transporters in restricting or permitting the brain penetration of various active ingredients of G. glabra. Bioavailability of glucose at brain level helps in its nourishment and proper functioning. A study on - Glucose, Learning and Memory [41, 42] by Psychology Professor Paul E. Gold found that as rats went through a maze, concentrations of glucose declined in the animal’s hippocampus, a key brain area involved in learning and memory – even more dramatically so in older brains. The study suggested that presence of optimal level of glucose in brain plays a vital role in learning and memory. The roots of G. glabra contain glycyrrhizin, which is a saponin that is 50 times

sweeter than sucrose, [10] Hence a suitable clinical study to evaluate the glucose level in brain after administration of glycyrrhizin and its role in learning and memory in healthy and diseased subjects would be an important milestone. Glabridin (Fig.2), liquiritigenin (Fig 4) and Isoliquiritigenin (Fig 5), the lipophilic compounds present in G. glabra with comparatively low-molecular weight (200 ∼ 700) are likely to be absorbed into blood and distributed to brain according to Lipinski rule of 5. [43] Liquiritigenin, a plant-derived highly selective estrogen receptor β agonist has been identified to alleviate the cognitive recession in the elder. [44] Glabrene also could be beneficial to increase memory due to estrogen-like activities, like isoliquiritigenin, liquiritigenin,

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GJRMI, Volume 1, Issue 6, June 2012, 247–257

and glabridin. [45, 46] Evaluating the role of these contents of G. glabra in improving cognitive functions in healthy and diseased subjects would be a significant avenue in future researches.

Impaired memory disorders like Dementia, Alzheimer’s and Learning disabilities of school going children, etc.

CONCLUSION

The author would like to acknowledge Prof M.S. Bhagel, Director of IPGT and RA, GAU, Jamnagar for encouraging in writing this article and Dr. Jyothy K B, PhD scholar, Dept of KB, IPGT and RA for helping me while preparing this work.

Glycyrrhiza glabra Linn (Yastimadhu) appears to be a promising herbal drug for cognitive improvement and it will be worthwhile to explore the potential of this noble drug with suitable clinical studies in the management of REFERENCES 1. Adams M, Gm¨under F, and Hamburger M (2007), “Plants traditionally used in age related brain disorders—a survey of ethnobotanical literature,” Journal of Ethnopharmacology, vol.113, no. 3, pp. 363– 381.

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diabetics". Proc. Natl. Acad. Sci. U.S.A. 101 (17): 6734–9. doi:10.1073/pnas.0306996101. PMC 404114. PMID 15071189. 30. Stewart, PM., Wallace, AM., Atherden, SM, Shearing CH and Edwards CRW. (1990) Clin. Sci.78, 49–54. Pmid : 2153495. 31. Raven J, Court J and Raven J (1977) Manual for Raven's Progressive Matrices and Vocabulary Scales (H. K. Lewis, London). 32. Lezak M (1995) Neuropsychological Assessment (Oxford Univ. Press, Oxford). 33. Wechsler D (1987) Wechsler Memory ScaleRevised (WMS-R) (Psychology Corporation, New York). 34. Wechsler D (1981) Manual of the Wechsler Adult Intelligence Scale-Revised (Psychology Corporation, New York). 35. Nelson H and Willison, J (1991) NART Test Manual (Part II) (NFER-Nelson, New York). 36. Matthews G (1990) British Journal of Psychology.81, 17–42. 37. Zigmond A and Snaith R (1983). Acta Psychiatr.Scand.67, 361–370. Pmid:6880820 38. Kadam Abhijit Ashok (2006), “A Study on the effect of Medhya Rasayana in School Going Children”, SDM College of Ayurveda, Hassan, Karnataka. 39. Neeta Patel (2008), “The conceptual and applied study of Medha and Manas in Ayurveda”, IPGT and RA, Gujarat Ayurved University, Jamnagar. 40. Charaka Samhitha with Ayurveda dipika commentary by Chakrapanidatta (Vimana sthana– 4/8) Chaukamba surbharti prakashana- Varanasi-2009.

Source of Support: Nil

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Conflict of Interest: None Declared

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