Immunological Evaluation In Vitro Of Polysaccharides And Triterpene Extracted From Ganoderma Lucidum

Objective:In this project, the polysaccharide and triterpenes of Ganoderma lucidum were studied as the research objects.Evaluating their effect on NK-92MI cells and Jurkat cell proliferation and apoptosis which will lay the foundation for further screening of effective parts of the Yi Yuan Kang.Methods:(1) Purificating the polysaccharide and triterpenes of Ganoderma lucidum sample solution according to the optimum selection of extraction, and then,prepare different concentrations of polysaccharides and triterpenes of Ganoderma lucidum sample solution and determine its purity. (2)NK-92MI cell and Jurkat cell growth curve was determined by tetrazolium colorimetric method (MTT), the OD value was as the index plate to screened the best concentration of NK-92MI cells and Jurkat cells. (3) The proliferation effect of the polysaccharide and triterpenes of Ganoderma lucidum on NK-92MI cells and Jurkat cells was determined by MTT assay, the OD value was as the index of the research of different concentration of the polysaccharide and triterpenes of Ganoderma lucidum NK-92MI cells and Jurkat cells proliferation. (4)The polysaccharide and triterpenes of Ganoderma lucidum of NK-92MI cells and Jurkat cell apoptosis was determined by Annexin V-FITC/PI double staining using flow cytometry to investigate effects of different concentration of the polysaccharide and triterpenes of Ganoderma lucidum of NK-92M1 cells and of Jurkat cells apoptosis, and analyzed by flow Jo software.Results:(1) The purity of the polysaccharide was 44.31%, the triterpenes purity was 40.44%. (2)It was conducive to cell growth when the inoculation concentration of NK-92MI cells was 4×105?8×105/ml and the inoculation concentration of Jurkat cells was 2×105?4×105/ml. (3) NK-92MI cell proliferation rate was maximum when the concentration of polysaccharide was 300?g/ml and cell dealed with polysaccharide for 12h, the proliferation rate was 20.594%,under this condition, polysaccharide extracted from Ganoderma lucidum had a better effect on NK-92MI cells; the proliferation rate of Jurkat cells was maximum reached 8.658% when the concentration was 400?g/ml and dealed with drugs for 24h, at thismoment,polysaccharide extracted from Ganoderma lucidum had a better effect on Jurkat cells. The proliferation rate of NK-92MI cells and Jurkat cells was maximum reached 24.023% and 7.430% when the concentration of triterpene was 1?g/ml and the reaction time was 24h and 12h. (4) The apoptosis inhibition ratio of NK-92MI. cells was maximum reached 30.75% when the concentration of polysaccharide was 300?g/ml and cell dealed with polysaccharide for 12h, at this moment, polysaccharide had better inhibitory effect on NK-92MI cells;.The Jurkat cell apoptosis inhibition rate was largest reached 13.01% when the concentration of polysaccharide was 400?g/ml and cell dealed with polysaccharide for 12h. The apoptosis inhibition rate of NK-92MI cells and Jurkat cells was maximum reached 53.36% and 18.45% when the concentration of triterpene was 1?g/ml and the reaction time was 12h and 48h, in this condition, triterpenes extracted Ganoderma lucidum had a better apoptosis inhibitory effect on Jurkat cell.Conclusion: (1)The polysaccharide and triterpenes extracted from Ganoderma lucidum which have suitable concentration can promote the effect of NK-92MI cells and Jurkat cells proliferation, but too large concentration maybe inhibit the cell proliferation. (2) The polysaccharide and triterpenes extracted from Ganoderma lucidum which have suitable concentration can inhibition the effect of NK-92MI cells and Jurkat cells apoptosis.