Effect Of Astragalus Polysaccharides On The Expression Of Tumor Necrosis Factor-? In Adipocytes

BackgroundDiabetes is a chronic endocrine disease,which is caused by many factors and characterized by high blood sugar in metabolism.long latency,more complications,extensive involvement of the organs as well as complicated clinical manifestations are the main pathological changes.With the development of national economy,increasing pace of life as well as reconstruction of the original life system,the incidence of diabetes is getting higher and higher.TNF-?(Tumor Necrosis Factor-?)is an important inflammatory mediator,which plays an important role in the pathogenesis of diabetes.Astragalus is a perennial legume,which is rich in saponins,alkaloids,polysaccharides,amino acids,selenium,zinc,copper and other trace elements.Astragalus polysaccharide is one of the main active components of Astragalus,which can be used as an immune promoter or regulator,as well as playing an important role in anti-virus,anti-tumor,anti-aging,anti-radiation,anti-stress,anti-oxidation and so on.ObjectiveIn this study,we tried to clarify whether Astragalus Polysaccharide has a certain infulence on the therapy of Metabolic diabetes Mellitus by observing the effect of Astragalus Polysaccharides on the expression of TNF-? in the adipocytes.This study used the adipocytes induced by adipocyte precursors as the research objects to explore the effect of Astragalus Polysaccharides on the expression of tumor necrosis factor-? by adopting MTT,ELISA,RT-PCR mehhods and so on,providing experimental data for the prevention and treatment of diabetes as well as clinical trials.Methods1.Preadipocytes were induced and cultured,and related changes in cytoplasm of the differentiated preadipocytes were observed by using red O staining.2.Chemosensitivity test: Adipocytes were cultured in the medium containing APS at different concentrations(0 ?g/ml as a control,10 ?g/ml,100 ?g/ml,500 ?g/ml,1000?g/ml).As has been reported,the OD values of the test cells at different time points(0 h,12 h,24 h,48 h)were detected to determine the optimal3.Effect of APS on the expression of TNF-? in adipocytes: At different time points(0d,1 d,2 d to 8 d),cell ELISA method was used to detect the expression of TNF-? in the medium and the adipocytes treated by the APS at the concentrations of 0 ?g/ml and100?g/ml,which 0 ?g/ml APS acted as the control and 100 ?g/ml APS acted as the experimented group.The expression of TNF-? mRNA was also detected in this two groups by RT-PCR.Result1.Compared with the other group,100 ?g/ml APS had an obvious effect on the inhibition of adipocytes.APS at the concentration of 100 ?g/ml strongly inhibited the growth of the adipocytes.2.Compared with the control,the expressions of TNF-? in experimental group began to decline after 3d both in the medium and in the adipocytes,while no obvious diffenences were observed at 0 d,1 d and 2 d.3.The levels of TNF-? mRNA both in the medium and the adipocytes showed no obvious differences between the experimental group and the control group during the first two days.However,three days later,mRNA level began to decline,which was consistent with our previous results.Conclusion100 ?g/ml is the optimal inhibition concentration of APS for adipocytes,and can obviously reduce the expressions of TNF-? in adipocytes,showing that APS is helpful to reduce the inflammatory reaction.