目的:探讨自噬抑制剂3-甲基腺嘌呤(3-MA)在H2O2诱导的神经胶质瘤U251细胞损伤过程中的作用.方法:实验分为4组:正常对照组、10 mmol/L 3-MA组、1 mmol/L H2O2组、1 mmol/L H2O2 +10 mmol/L 3-MA组.MTT法检测各组的U251细胞增殖率;MDC染色检测细胞自噬空泡的变化;Hoechst 33342染色检测细胞核染色质凝聚变化;流式细胞术检测细胞凋亡率.结果:与对照组相比,单独应用3-MA对U251细胞无明显影响.H2O2作用组U251细胞增殖率明显降低,细胞内出现自噬空泡,细胞核染色质凝聚,细胞凋亡率增加.3-MA与H2O2联合作用于U251细胞时,与单独应用H2O2组相比,抑制了H2O2引起的胞内自噬空泡的积聚,但细胞凋亡率明显增加.结论:自噬抑制剂3-MA能够一定程度地抑制H2O2诱导的U251细胞自噬,但促进了细胞凋亡,表明自噬在H2O2诱导的神经胶质瘤U251细胞损伤过程中很可能起到保护性作用.%AIM: To investigate the role of autophagy inhibitor 3 - methyladenine( 3 - MA ) in the injury of U251 glioma cells induced by H2O2.METHODS: The following groups in this study were set up: control group, 10 mmol/L 3 -MA group, 1 mmol/L H2O2 group and 1 mmol/L H2O2 +10 mmol/L 3 - MA group.The viability of U251 cells in each group was detected by MTT assay.Autophagic vacuoles in the cells were observed by staining with MDC.The cells were stained with Hoechst 33342 to determine the chromatin condensation.Cell apoptotic ratio was measured by flow cytometry analysis.RESULTS: Compared with control group, no effect of 3 - MA on the viability of U251 cells was observed.In H2O2 group, the cell viability decreased and cell apoptotic ratio increased.The autophagic vacuoles and nuclear chromatin condensation in the cells were also detected.Compared with H2O2 group, addition of 3 - MA inhibited the increase in autophagic vacuoles but exacerbated the apoptosis.CONCLUSION: Autophagy inhibitor 3 - MA inhibits autophagy partially, but exacerbates apoptosis in U251 cells, indicating that autophagy exerts protective effect in the process of injury in U251 cells induced by H2O2.
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