摘要
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 P 20 were detected with Western blotting assay. The results showed that during the process of DA induced apoptosis, the cleaved caspase 3 P 20 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 didnt 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.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2002年第4期621-624,共4页
Progress In Biochemistry and Biophysics
基金
福建省自然科学基金重点课题资助项目 (C982 0 0 4)
福建省科技厅重点课题资助项目 (2 0 0 1Z0 3 7)~~
