【摘要】 目的研究黄芪甲苷通过激活细胞自噬抑制缺氧缺糖/复氧复糖诱导PC12细胞凋亡的作用。方法取对数生长期的PC12细胞,随机分为7组:正常对照组、模型组、溶媒组、黄芪甲苷组、黄芪甲苷+自噬抑制剂组、自噬抑制剂组、自噬激活剂组。除正常对照组外,其余各组剥夺氧、糖3h后再复氧、复糖12 h,并于复氧、复糖的同时给予相应药物处理。用透射电镜观察自噬小体的变化,Western blot检测LC3-II、Beclin1、半胱氨酸天冬氨酸蛋白酶3(Caspase3)的表达,流式细胞术和TUNEL染色检测细胞凋亡。结果与正常对照组相比,模型组出现典型的自噬小体,LC3-II、Beclin1表达均增多(P<0.05),同时Caspase3表达、细胞凋亡率和凋亡指数也升高(P<0.05)。与模型组相比,黄芪甲苷组、自噬激活剂组自噬小体数量、LC3-II和Beclin1表达均增多(P<0.05),Caspase3表达、细胞凋亡率和凋亡指数均下降(P<0.05);而自噬抑制剂组自噬小体数量、LC3-II和Beclin1表达减少(P<0.05),Caspase3表达、细胞凋亡率和凋亡指数升高(P<0.05);溶媒组与模型组之间无显著差异(P>0.05)。与黄芪甲苷组相比,黄芪甲苷+自噬抑制剂组、自噬抑制剂组自噬小体数量、LC3-II和Beclin1表达均减少(P<0.05),Caspase3表达、细胞凋亡率和凋亡指数均升高(P<0.05)。结论黄芪甲苷可通过激活细胞自噬而抑制缺氧缺糖/复氧复糖诱导的PC12细胞凋亡,从而发挥神经保护作用。更多还原

【Abstract】 Objective To study the inhibitory effect of astragaloside IV( AST) on the apoptosis of PC12 cells induced by oxygen-glucose deprivation/oxygen-glucose rehabilitation( OGD/OGR) through activating autophagy. Methods PC12 cells in logarithmic phase were collected and randomly divided into normal group,OGD/OGR group( model group),AST solvent control group( DMSO group),AST group,AST + autophagy inhibitor group( AST + 3-MA group),3-MA group and autophagy activator group( RAPA group). Except of normal group,all other groups were given treating of OGD for 3 h and then OGR for 12 h and simultaneous medication treating. The morphological changes of autophagosomes were observed by using transmission electron microscope( TEM),expressions of LC3 II,Beclin1 and Caspase-3were detected by using Western blotting,and apoptosis of PC12 cells were detected by using flow cytometry after TUNEL staining. Results Compared with normal group,the typical autophagosomes were observed,expressions of LC3 II,Beclin1 and Caspase3 increased,and apoptosis rate and apoptosis increased( P < 0. 05). Compared with model group,the quantity of autophagosomes and expressions of LC3 II and Beclin1 increased( P < 0. 05),and expression of Caspase3 and apoptosis rate and apoptosis decreased in AST group and RAPA group( P < 0. 05). Compared with model group,the quantity of autophagosomes and expressions of LC3 II and Beclin1 decreased( P < 0. 05),and expression of Caspase3 and apoptosis rate and apoptosis increased in 3-MA group( P < 0. 05),while DMSO group and model group had no significant difference( P > 0. 05). Compared with AST group,the quantity of autophagosomes and expressions of LC3 II and Beclin1 decreased( P < 0. 05),and expression of Caspase3 and apoptosis rate and apoptosis increased in AST + 3-MA group and 3-MA group( P < 0. 05). Conclusion AST play a neuroprotective role through activating autophagy and inhibiting apoptosis of PC12 cells induced by OGD/OGR.更多还原