Study On Effects Of Polysaccharide From Allium Mongolicum Regel On Regulation Of Immunologic Functions Of Peripheral Blood Lymphocytes In Sheep

This paper conducded to studied the effect of allium mongolicum regelpolysaccharideson regulation of immunity function of peripheral blood lymphocyte insheep by in vitro. The contents and results as follows:1.Sheep peripheral blood lymphocytes were isolated and cultured aseptically invitro, and the morphology of lymphocytes that were cultured for observed underinverted phase contrast microscope. The observation shows that the cell bodies arecircular, small, with uniform size, and grow suspended in the culture medium.2.With MTT colorimetric assay to measured lymphocyte proliferation indifferent allium mongolicum regel polysaccharides concentration (0-250μg/mL) anddifferent functioning time(24h,48h,72h). The result shows that allium mongolicumregel polysaccharides promoted the proliferation of peripheral blood lymphocyte insheep and the effect was in a time dependent manner and dose dependent manner. Theproliferation of lymphocytes reached the highest level when the time of action was24hours and the concentration was250μg/mL.3.The influence of allium mongolicum regel polysaccharides on sheep peripheralblood T lymphocyte subsets was observed using flow cytometry. The result revealedthat allium mongolicum regel polysaccharides could increase the number of sheepperipheral blood CD4+cell and CD4＋/CD8＋ratio, and reduce the number of CD8＋cell.4.Radioimmunoassay was used to test the effect of allium mongolicum regelpolysaccharides with different concentrations (0-250μg/mL) on IL-6which was in theculture medium supernatant of sheep peripheral blood lymphocytes at differentfunctioning time (12h,24h,48h,72h). The result indicated that allium mongolicumregel polysaccharides promoted the secretion of IL-6of sheep peripheral bloodlymphocytes. When the funtion time continued for24hours the secretion level of IL-6induced by ConA reached its highest point.5.The technique of Real-Time PCR was applied to test the effect of alliummongolicum regel polysaccharides on mRNA expression of IL-6of sheep peripheral blood lymphocytes. The result indicated that each concentration group of alliummongolicum regel polysaccharides showed promotion on mRNA expression of IL-6attime points12h,24h,48h, and72h. At the concentration of50μg/mL, there was nosignificant (p>0.05) promotion on ConA induced lymphocyte IL-6mRNA expressionlevels at all the time points. With the lengthening of funtion time, the ConA inducedIL-6mRNA expression level reduced gradually. At the functioning time of24hoursand allium mongolicum regel polysaccharides concentration of200μg/mL, thereshowed very significant (p<0.001) promotion on ConA induced lymphocyte IL-6mRNA expression.