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目的 探讨PC12细胞缺氧后细胞凋亡与DNA损伤的关系。方法 采用TUNEL法 ,结合应用流式细胞术观察PC12细胞缺氧培养不同时间点细胞凋亡现象 ,以及DNA损伤相关基因表达的改变。结果 在缺氧 0 .5h开始出现凋亡细胞 ,此时P5 3、P2 1waf1/cip1蛋白表达开始增高。至缺氧 1h凋亡细胞达高峰 ,此时P5 3、P2 1蛋白表达最高 (P <0 .0 5 )。至缺氧 6～ 12h ,则以坏死为主 ,此时以上基因表达均减弱。结论 PC12细胞在缺氧早期 (0 .5～ 2h)主要出现以凋亡为主的细胞死亡 ,伴随DNA损伤相关基因表达的动态改变 ,提示缺氧后PC12细胞凋亡部分是由于DNA损伤严重、损伤不能及时修复所致。
Objective To investigate the relationship between apoptosis and DNA damage in PC12 cells after hypoxia. Methods TUNEL method was used in combination with flow cytometry to observe the apoptosis of PC12 cells at different time points and the changes of gene expression related to DNA damage. Results Apoptotic cells began to appear at 0.5 h of hypoxia, and the expression of P5 3 and P2 1waf1 / cip1 began to increase. The apoptotic cells reached the peak at 1 hour hypoxia, at which point P5 3 and P2 1 protein expression was the highest (P <0.05). To hypoxia 6 ~ 12h, then the main necrosis, at this time more than the gene expression were weakened. Conclusion The apoptosis of PC12 cells mainly occurs in the early stage of hypoxia (0.5-2h), with the dynamic changes of DNA damage-related gene expression. It is suggested that the apoptosis of PC12 cells after hypoxia is partly due to the severe DNA damage, Damage can not be repaired promptly.