Effects And Mechanism Of3’-Methoxypuerarin Against H₂O₂-induced Oxidative Damage In INS-1Cells

Objective: To investigate the effects of3’-Methoxypuerarin on proliferation, insulin secretionand protection on oxidative damage induced by H2O2of INS-1cells and their possiblemechanism.Methods: INS-1β cell was cultured in vitro. Set control group, Medium group, model group,Positive drug group and drug groups(0.3,1,3,10,30μmol/L). INS-1cells were treated for48h, and the proliferation of INS-1cells was tested by MTT assay. It is established that theInsulin Secretion Model by using5.6、16.7mmol/L glucose, and tested the insulin secretionvia RIA. The oxidative stress cell model was established with H2O2, the cell viability wastested by MTT assay. Morphological changes of oxidative damage of INS-1cells wereobserved by Hochest33258staining. The levels of intracellular ROS and the apoptosis werequantified by fluorescence with DCF-DA and AnnexinⅤ/PI. The mRNA levels of Bcl-2andBax in oxidative damage of INS-1cells were measured by Q-PCR.Results:1、Effects on Proliferation of INS-1CellsCompared with Medium group,3’-Methoxypuerarin have promotion in0.3~10μmol/L(P<0.05), in which it could reach18.08%in1μmol/L.2、Effects of3’-Methoxypuerarin against H2O2-induced Oxidative Stress in INS-1CellsCompared with model group,3’-Methoxypuerarin have protection effect in0.3~3μmol/L(P<0.01or P<0.05), and could increase the proportion of normal cells in0.1~1μmol/L with the increase of drug concentration, also significantly decrease intracellularROS levels in0.1~1μmol/L(P<0.01or P<0.05), and reduced apoptosis significantly in0.3~1μmol/L(P<0.01or P<0.05).3、Effect of BIS and GSIS of H2O2-challenged INS-1CellsCompared with model group,3’-Methoxypuerarin have a significant role of BIS, GSISin0.1~1μmol/L(P<0.01or P<0.05).4、Mechanism of Bcl-2、Bax in H2O2-challenged INS-1CellsCompared with model group,3’-Methoxypuerarin up-regulation of Bcl-2mRNAexpression in0.1~1μmol/L，and down-regulate Bax mRNA in1μmol/L(P <0.01). Conclusions:1、3’-Methoxypuerarin significantly promote the proliferation, could improve cell viabilityand insulin secretion of H2O2-challenged INS-1cells. In which3’-Methoxypuerarin couldreduce apoptosis, inhibit intracellular ROS;2、3’-Methoxypuerarin protect β cells via up-regulation of Bcl-2gene expression anddown-regulation of Bax mRNA expression.