Kong Xiao-Xia, Zhang Hong-Yu, Chen Ran, Fan Xiao-Fang, Gong Yong-Sheng
Institute of Hypoxia Medical Research, Teaching Center of Functional Experiment, Wenzhou Medical College, Wenzhou 325035, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2011 Nov;27(4):471-4.
To investigate the time order of autophagy and apoptosis in human U251 cells injury after H2O2 treatment.
4 groups in this study were set up, normal control group, 1 mmol/L H2O2 (6 h,12 h, 24 h) group. The viability of U251 cells treated with H2O2 was measured by MTT assay. Cell apoptotic ratio was determined by flow cytometry analysis. Autophagic vacuoles were stained with monodansylcadaverine. The protein level of Beclin 1 and cytosolic cyt c were assayed by using Western blot.
Compared with the control group, cell viability decreased significantly under 1 mmol/L H2O2 treatment in time-dependent way. Autophagic vacuoles and the expression of autophagic protein Beclin 1 increased at 6 h, but cell apoptotic ratio and cytosolic cyt c protein did not change obviously, cell apoptotic ratio and cytosolic cyt c protein level increased at 12 h and 24 h (P < 0.05).
Oxidative stress induced autophagy and apoptosis in U251 glioma cells, and autophagy eventuated ahead of apoptosis.
探讨过氧化氢（H2O2）处理后人U251细胞损伤过程中自噬与凋亡的时间顺序。
本研究设置4组，即正常对照组、1 mmol/L H2O2（6小时、12小时、24小时）组。采用MTT法检测H2O2处理后U251细胞的活力。通过流式细胞术分析测定细胞凋亡率。用单丹磺酰尸胺对自噬空泡进行染色。采用蛋白质印迹法检测Beclin 1和胞质细胞色素c的蛋白水平。
与对照组相比，1 mmol/L H2O2处理下细胞活力呈时间依赖性显著降低。自噬空泡及自噬蛋白Beclin 1的表达在6小时时增加，但细胞凋亡率和胞质细胞色素c蛋白无明显变化，细胞凋亡率和胞质细胞色素c蛋白水平在12小时和24小时时升高（P < 0.05）。
氧化应激诱导U251胶质瘤细胞发生自噬和凋亡，且自噬先于凋亡出现。
