黄芪甲苷调控自噬减轻氧糖剥夺/复氧复糖PC12细胞氧化应激损伤研究

Study of astragaloside Ⅳ alleviating oxidative stress injury of PC12 cells induced by oxygen-glucose deprivation/reoxygenation through regulating autophagy

目的观察黄芪甲苷对氧糖剥夺/复氧复糖PC12细胞自噬和氧化应激的影响。方法将PC12细胞分为正常组、模型组(即氧糖剥夺/复氧复糖组)、黄芪甲苷组、自噬抑制剂+黄芪甲苷组。用CCK-8法检测细胞活性;ELISA法检测MDA含量、SOD和GSH-Px活性;透射电镜和MDC荧光染色检测自噬小体;Western blot检测Beclin1蛋白表达。结果与正常组相比,模型组细胞活性降低(P<0.05),MDA含量增多、SOD和GSH-Px活性下降(P<0.05),可见自噬小体,Beclin1表达增多(P<0.05);与模型组相比,黄芪甲苷组细胞活性升高(P<0.05),MDA含量减少、SOD和GSH-Px活性升高(P<0.05),自噬小体数量增多(P<0.05),Beclin1表达增多(P<0.05);当给予自噬抑制剂处理时,自噬抑制剂在减轻自噬的同时,可明显拮抗黄芪甲苷的抗氧化作用。结论黄芪甲苷可通过上调自噬减轻氧糖剥夺/复氧复糖诱导的PC12细胞氧化应激损伤,发挥保护作用。

Aim To observe the effects of astragalosideⅣon autophagy and oxidative stress induced by oxy-gen-glucose deprivation/reoxygenation in PC12 cells.Methods PC12 cells were divided into normal group,model group(oxygen-glucose deprivation/reoxygenation group),astragalosideⅣgroup,and autophagy inhibitor+astragalosideⅣgroup.CCK-8 was used to detect cell viability,and ELISA to detect MDA,SOD and GSH-Px.Transmission electron microscope and MDC fluorescent staining were employed to observe the changes of autophagosome,and Western blot to detect the protein expression of Beclin1.Results Compared with normal group,the cell activity in model group decreased(P<0.05),MDA content increased,SOD and GSH-Px activity decreased(P<0.05),and autophagosomes could be seen and the protein expression of Beclin1 increased(P<0.05).Compared with model group,the cell activity in astragalosideⅣgroup increased(P<0.05),MDA content decreased,SOD and GSH-Px activity increased(P<0.05),the number of autophagosomes and the protein expression of Beclin1 increased(P<0.05).When autophagy inhibitor was given at the same time,the autophagy inhibitor could obviously antagonize the antioxidant effect of astragalosideⅣwhile alleviating autophagy.Conclusions AstragalosideⅣcan protect PC12 cells from oxidative stress injury induced by oxygen-glucose deprivation/reoxygenation by up-regulating autophagy.