自噬对不同低氧条件下PC12细胞存活的影响  

The effects of autophagy on cell survival under different hypoxia

作　　者：何云凌[1] 吴丽颖[1] 黄欣[1] 赵彤[1] 丁学锋[1] 吴奎武[1] 范明[1] 朱玲玲[1] 

机构地区：[1]军事医学科学院基础医学研究所认知科学研究室,北京100850

出　　处：《中国应用生理学杂志》2013年第3期193-196,I0001,共5页Chinese Journal of Applied Physiology

基　　金：国家973计划资助项目(2012CB518200;2011CB910800);国家自然科学基金资助项目(81071066;81000856;31271211)

摘　　要：目的:探讨不同低氧对细胞存活和自噬的调节作用。方法:采用不同氧含量处理大鼠肾上腺嗜铬瘤细胞(PC12),MTT方法检测细胞存活情况;Western blot检测自噬特异性标记物微管相关蛋白轻链3(LC3)和接头蛋白p62的表达变化;流式细胞分析测定低氧及阻断自噬后活性氧水平(ROS)的变化情况。结果:不同低氧细胞存活能力不同:适度低氧促进细胞存活,严重低氧导致细胞存活能力下降;自噬标记蛋白在不同低氧下表达水平不同:适度低氧使LC3-II和p62表达增加,严重低氧则反之,LC3-II和p62表达降低;与常氧相比,适度低氧没有改变ROS水平,严重低氧使ROS水平增加;用自噬抑制剂3-MA抑制自噬后,各种氧含量下的ROS水平都被升高,其中,适度低氧与严重低氧ROS升高幅度明显高于常氧情况。结论:适度低氧促进细胞存活,严重低氧导致细胞死亡,自噬活性介导了不同低氧的作用效果。Objective：To investigate the regulation of different hypoxia on cell survival and autophagy.Methods： PC12 cells were treated with different hypoxia.The cell survival was measured by MTT assay,expressions of LC3 and p62 were marked for autophagy detected by Western Blot,and the level of reactive oxygen species（ROS）was analyzed by flow cytometry.Results： The cell viability was different under different hypoxia：moderate hypoxia promoted cell viablity,and severe hypoxia caused a decrease in cell viability;autophagy marker molecules,p62 and LC3-II expressions were different：moderate hypoxia increased p62 and LC3-II expressions,in contrast,severe hypoxia led to the decrease of p62 and LC3-II expressions;compared to normoxia,moderate hypoxia did not change the levels of ROS,while severe hypoxia increased the levels;3MA,the inhibitor of authophagy,elevated the levels of ROS in the three oxygen concentrations,additionally,the increased amplitudes in the moderate and severe hypoxia groups were higher than that in the normoxia group.Conclusion： Moderate hypoxia promotes cell survival,severe hypoxia causes the cell death,and the autophagy activity may mediate the effects of different hypoxia.

关 键 词：低氧 自噬 活性氧 LC3II P62 

分 类 号：Q251[生物学—细胞生物学] R364.4[医药卫生—病理学]