Study Of Echinops Latifolius Tausch Polysaccharide B On Oxidative Damage,Apoptosis And Insulin Secretion Effect In INS-1 Cells

The previous experiments showed that the Echinops Latifolius Tausch Polysaccharide B?ETPB?has protective effect on pancreatic injury,increase insulin secretion and improve insulin resistance in diabetic model animals.In this experiment,INS-1 islet beta cells were cultured to explore the effect of ETPB directly on islet beta cells,and to observe its effects on cell oxidative damage,apoptosis and insulin secretion,and to provide a theoretical basis for the related research on the prevention and control of T2DM.The results show that:1.The MTT method was used to determine the maximum safety concentration of ETPB 250?g/mL,and ETPB action 48h could increase the cell survival rate compared with the H₂0₂ model group,and improve the SOD activity and reduce the MDA content.2.The results of AO-EB staining after ETPB treatment showed that the apoptotic cells in ETPB group were less than that of the model group,and the expression of mRNA by using RT-qPCR could improve the expression of Bcl-2 and decrease the expression of Bax.Using Western Blot to detect related protein expression can increase Bcl-2 expression and decrease Bax,Cytc and Caspase-3 expression.3.After ETPB treatment,radioimmunoassay was used to detect the secretion of insulin,which could improve the glucose stimulation of insulin secretion,which had obvious effect on the normal sugar culture of BIS,and the effect on high glucose culture of GSIS was obvious,and the expression of PDX-1 and IRS-2 could be increased at the level of mRNA.Conclusion:1.ETPB can protects against oxidative damage by increasing SOD activity and decreasing MDA content.2.ETPB can regulate mitochondrial apoptosis pathway related proteins to inhibit H₂0₂ induced apoptosis.3.ETPB can increase glucose stimulated insulin secretion and increase the expression of PDX-1 and IRS-2mRNA.