福建省自然科学基金重点课题资助项目(C982004)和福建省科技厅重点课题资助项目(2001Z037).
This work was supported by a key grant from The Nature Science Foundation of Fujian Province (C982004) and a Key Program of Fujian Science and Technology Bureau (2001Z037).
通过末端脱氧核苷酸转移酶介导dUTP缺口翻译法和DNA凝胶电泳观察多巴胺(DA)对PC12细胞凋亡的诱导作用, 并经蛋白质印迹法检测胞浆细胞色素c、Bcl-2和Bax蛋白以及活化型半胱氨酸蛋白酶3(caspase-3)水平. 结果表明, 在DA诱导PC12细胞凋亡的过程中, 可见PC12细胞中活化型caspase-3蛋白表达, 胞浆中细胞色素c水平明显增高, 同时Bcl-2蛋白水平下降, 而Bax蛋白水平明显增加. 环孢菌素A预处理对细胞色素c释放和caspase-3激活有明显的抑制作用, 而对Bcl-2和Bax蛋白影响不明显. 结果提示, Bcl-2和Bax蛋白、细胞色素c以及caspase-3可能参与DA诱导PC12细胞凋亡, 线粒体细胞色素c向胞浆释放可能是其中的中心环节.
After treatment of PC12 cells with various doses of dopamine (DA), the apoptotic cells were observed with TdT-mediated dUTP nick end labelling method (TUNEL) and DNA gel electrophoresis. Meanwhile, Bcl-2, Bax and cytosolic cytochrome c as well as cleaved caspase-3 P20 were detected with Western blotting assay. The results showed that during the process of DA-induced apoptosis, the cleaved caspase-3 P20 and the release of cytochrome c into cytosol were elevated markedly compared with the controls. The protein levels of Bcl-2 were decreased, however, Bax protein was increased significantly. After pretreatment of PC12 cells with cyclosporine A at 1 μmol/L for 24 h, the release of cytochrome c and caspase-3 activation were blocked almost. But it didn't show effect on protein levels of Bcl-2 and Bax. The results suggested that Bcl-2 family and cytochrome c and caspase-3 might be involved in the progression to apoptosis induced by DA, furthermore, the release of cytochrome c from mitochondria might be an essential event.
朱元贵,陈晓春,陈丽敏,方芳,周宜灿,赵朝辉.细胞色素c的释放是多巴胺诱导PC12细胞凋亡的重要环节[J].生物化学与生物物理进展,2002,29(4):621-624
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