主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2017, Vol. 40 ›› Issue (5): 405-412.doi: 10.3969/j.issn.1006-2157.2017.05.009

• 科技之窗 • 上一篇    下一篇

黄芪甲苷通过激活自噬抑制缺氧缺糖/复氧复糖诱导的PC12细胞凋亡*

靳晓飞1, 周晓红2, 武密山2, 张颖2, 赵艳萌2, 李媛1, 吴增1, 高维娟1,2#   

  1. 1 承德医学院病理生理学教研室 河北 067000;
    2 河北中医学院 河北省心脑血管病中医药防治重点实验室
  • 收稿日期:2016-11-20 出版日期:2017-05-30 发布日期:2017-05-30
  • 通讯作者: 高维娟,女,博士,教授,博士生导师,研究方向:缺血性脑血管病的发生机制及中医药防治,E-mail:gwj6088@163.com
  • 作者简介:靳晓飞,男,在读硕士生
  • 基金资助:
    *河北省应用基础研究计划重点项目(No.16967756D),河北省普通高校高层次人才科学研究计划项目(No.GCC2014031),河北省博士学位点科研能力建设项目(No.169677128D),河北省研究生创新资助项目(No.冀学位[2016]3号)

Inhibitory effect of astragaloside IV on PC12 apoptosis induced by oxygen-glucose deprivation/oxygen-glucose rehabilitation through activating autophagy*

JIN Xiaofei1, ZHOU Xiaohong2, WU Mishan2, ZHANG Ying2, ZHAO Yanmeng2, LI Yuan1, WU Zeng1, GAO Weijuan1,2#   

  1. 1 Department of Pathophysiology, Chengde Medical College, Hebei 067000, China;
    2 Hebei University of Traditional Chinese Medicine, Hebei Key Laboratory of Chinese Medicine Research on Cardio-Cerebrovascular Disease, Hebei 050091, China
  • Received:2016-11-20 Online:2017-05-30 Published:2017-05-30
  • Supported by:
    Planned Key project of Applied Basic Research in Hebei Province (No.16967756D), Planned Project of High-level Talents in Ordinary Universities in Hebei Province (No.GCC2014031), Project of Construction of Capacity for Scientific Research in Doctor’S Degree Authorization Centers in Hebei Province (No.169677128D), Project of Supporting graduate Innovation in Hebei Province (No. Hebei academic degree[2016]3)

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3 h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。

关键词: 黄芪甲苷, 细胞自噬, 细胞凋亡, PC12细胞, 缺氧缺糖/复氧复糖

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 LC3II, Beclin1 and Caspase-3 were 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 LC3II, Beclin1 and Caspase3 increased, and apoptosis rate and apoptosis increased (P<0.05). Compared with model group, the quantity of autophagosomes and expressions of LC3II 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 LC3II 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 LC3II 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.

Key words: astragaloside, autophagy, apoptosis, PC12 cells, oxgen-glucose deprivation/oxgen-glucose rehabilitation

中图分类号: 

  • R285.5