上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

低氧引起PC12细胞保护性自噬的激活

汪文英，崔德荣，江伟   

  1. 上海交通大学附属第六人民医院麻醉科， 上海 200233
  • 出版日期:2015-08-28 发布日期:2015-09-30
  • 作者简介:汪文英（1989—）， 女， 硕士生; 电子信箱: wenying_wang@163.com。
  • 基金资助:

    国家教育部博士点基金（20120073110087）

Activation of protective autophagy of PC12 cells induced by hypoxia

WANG Wen-ying, CUI De-rong, JIANG Wei   

  1. Department of Anesthesiology, the Sixth People's Hospital,  Shanghai Jiao Tong University, Shanghai200233, China
  • Online:2015-08-28 Published:2015-09-30
  • Supported by:

    Doctoral Fund of Ministry of Education of China，20120073110087

PDF (PC)

1327

摘要：

目的  初步探讨低氧对PC12细胞自噬功能的影响。方法  应用低氧培养箱给予实验组细胞1%氧浓度的低氧处理，对照组置于正常含氧的培养箱中。首先，实验组细胞分别低氧处理6、12、24 h，用Western blotting检测自噬相关蛋白LC3和Beclin1的表达，用RT-PCR检测LC3 mRNA的变化，用透射电子显微镜（透射电镜）检测自噬小体的变化；其次，给予自噬抑制剂3-MA后，用MTT检测细胞活力，用caspase3活性检测试剂盒检测caspase-3的活性，用Western blotting方法检测cleaved caspase-3 的表达。结果  ①随着低氧时间延长，PC12细胞自噬相关蛋白LC3及Beclin1表达增高，12~24 h最高，随后下降，LC3 mRNA水平也出现相应增高趋势。②给予自噬抑制剂3-MA能进一步降低低氧处理后细胞活力，而caspase-3的活性增强，其活性形式表达增高。结论  自噬在PC12细胞发生低氧被激活，并且可能通过抑制凋亡发挥保护作用。

关键词: 肾上腺嗜铬瘤细胞, 低氧, 自噬

Abstract:

Objective  To explore the effect of hypoxia on the autophagy of PC12 cells. Methods  The hypoxia incubator was used to establish the hypoxic environment with oxygen concentration of 1% for treating cells of the experimental group, while cells of the control group were placed in the incubator with normal oxygen concentration. Firstly, cells of the experimental group were treated in hypoxic environment for 6, 12, and 24 h respectively. Expressions of autophagy-related proteins LC3 and Beclin1 were detected by Western blotting and changes of mRNA of LC3 were detected by RT-PCR. Changes of autophagosomes were observed by the transmission electron microscope. Secondly, after being treated by autophagy inhibitor 3-MA, the viability of cells was detected by MTT, the activity of caspase-3 was detected by caspase-3 activity assay kit, and the expression of cleaved caspase-3 was detected by Western blotting. Results  ①With the increase of time of exposure to hypoxic environment, expressions of autophagy-related proteins LC3 and Beclin1 and mRNA level of LC3 of PC12 cells increased, reached peaks during 12-24 h, and decreased afterward. ②The autophagy inhibitor 3-MA further decreased the viability of cells treated by hypoxic environment and increased the activity of caspase-3 and expression of active caspase-3. Conclusion  Hypoxia activates the autophagy of PC12 cells and may play a protective role by inhibiting the apoptosis.

Key words: PC12 cells, hypoxia, autophagy