Novel application of Mushroom Therapy in Management of Rare Auto-Immune Diseases Rashmi Thakare 1 and Rahul Suryawanshi 2 1 2
Wageningen University and Research Centre, The Netherlands.
College of Veterinary and Animal Sciences, Udgir, Dist. Latur, Maharashtra
RESULTS
ABSTRACT
Polysaccharide determination of mushroom extracts
Background: According to ORDI 450 rare diseases have been reported in India. The most common rare diseases are found to be autoimmune diseases(more than 100). Mushrooms have considerable immunomodulating potential. Rationale: The immunomodulating effect of the mushrooms Pholiotanameko (Nameko) and Lentinulaedodes (Shiitake) were measured for the cytokine production of THP-1 monocytes and macrophages after stimulation with full extracts and respective polysaccharide extractions. These polysaccharide extractions were examined because β-glucans are hypothesized to be major contributors in the immunomodulating function of mushrooms. Strategy: The THP-1 cells were stimulated with the mushroom extracts in the concentrations 100 µg/ml and 1000 µg/ml. The effect of β-glucan, the major contributors in the immunomodulating function of mushrooms was measured by using Dubois method. To check the effectiveness of β-glucan, comparatively the effect of the extracts without β-glucan was measured by blocking dectin-1, the protein thought to be the most important β-glucanreceptor. The quantification of the cytokine concentrations of IL-6 and IL-10 was done by Enzyme- Linked Immuno Sorbent Assay.
Figure 1: shows that β-glucan extractions indeed contain more polysaccharides than full mushroom extracts. The Nameko β-glucan extract (NβE) contains 14 mg/ml saccharide, more than three times the amount in the full extract (4 mg/ml). Shiitake shows a similar trend, its β-glucan extract (SβE) containing roughly twice as much as its full extract.
Result: The results showed that no cytokine production was found in the positive control of THP-1 monocytes, while in case of macrophages, IL-10 production was more with Nameko extracts than Shiitake extracts and IL-6 production was more with Shiitake extracts than Nameko extracts.
Cytokine production of macrophages stimulated with mushroom extracts
Conclusion: Mushrooms Pholiotanameko (Nameko) and Lentinulaedodes (Shiitake) have significant potential to induce immunomodulating effects. They can surely be thought as natural medicine for curing rare auto-immune diseases. Keywords: Mushrooms, immunomodulating potential, auto-immune diseases.
INTRODUCTION Figure 2: The sample numbers and concentrations represent (1)Nameko full mushroom extract, (2)Nameko β-glucan extract, (3) Shiitake full extracts and (4) Shiitake β-glucan extracts. Herein 100 and 1000 correspond to extracts of 100 μg/ml and 1000 μg/ml respectively. This graph shows that only macrophages stimulated with 1000 μl/ml β-glucan extracts produce IL-6, and that IL-6 concentrations are decreased when the dectin-1 receptor is blocked. The Shiitake extracts stimulate IL-6 production more than Nameko extracts.
Mushroom compounds have been reported to have profound immunomodulating effects in vitro and are predominant in cytokine production.
1.
Figure 3: The sample numbers and concentrations represent:(1)Nameko full mushroom extract, (2)Nameko β-glucan extract, (3) Shiitake full extracts and (4) Shiitake β-glucan extracts. Herein 100 and 1000 correspond to extracts of 100 μg/ml and 1000 μg/ml respectively. This graph shows that IL-10 concentrations are lower when the dectin-1 receptor is blocked, and that Nameko extracts stimulate the IL-10 production more than Shiitake extracts.
The cytokines that were monitored were interleukin 6 and 10 (IL-6 and IL-10) . IL-6 is proinflammatory cytokine while IL-10 is an antiinflammatory cytokine.
METHOD
Cytokine production of monocytes stimulated with mushroom extracts
Mushrooms
Proliferation test PMA induction
Shiitake
Determination Β-glucan
Whole extract
Nameko
Monocyt
Β-glucan extracts
Macrophage
Cells after supernatant is removed
CONCLUSION
IL-10 production significantly increased in macrophages inoculated with either Shiitake or Nameko extracts, as well as with their respective polysaccharide extractions.
Nameko was found to be more potent than Shiitake in stimulating IL-10 production. Dectin-1 receptor blocking showed that polysaccharide, including β-glucan, is a major factor in mushrooms’ IL-10 production enhancing effect.
These results show that Pholiota Nameko may be a valuable natural antiinflammatory mushroom, though its effects on other cytokines would have to be assessed and weighed against each other.
The data shows that mushroom extracts do not induce monocytes to produce cytokines.
Thus we can say that the two mushrooms choosed for study produces immunomodulatory cytokines that allows the immune system to respond appropriately to stressors and help reduce unwanted inflammations. Thereby this mechanism being useful to cure many rare autoimmune diseases.
Β-glucan receptor blocking with dectin 1antibody
Mushroom extracts added to cells
Harvest supernatant with cytokines
To determine the effect of mushroom extracts in THP-1 monocytes, ELISAs were used to detect the cytokine levels of TNF-α and IL-10. The results are not reported upon, as no cytokine production was found.
ELISA