缺氧复氧致神经细胞凋亡中Bcl-2、Bax基因蛋白的表达  

THE EXPRESSION OF Bcl-2 AND Bax PROTEIN IN THE APOPTOSIS OF NEURONS FOLLOWING HYPOXIA/REOXYGENATION

作　　者：季凤清[1] 岳旭[2] 孙海梅[1] 郭艳茹[2] 郭崇洁[1] 赵天德[2] 

机构地区：[1]首都医科大学组织学胚胎学教研室,北京100054 [2]中日友好临床医学研究所,北京100029

出　　处：《解剖学报》2001年第4期338-342,T011,共6页Acta Anatomica Sinica

基　　金：北京市科干局优秀青年骨干培养基金资助 (98 11)

摘　　要：目的　探讨缺氧复氧致原代培养的大鼠大脑皮层神经细胞凋亡中Bcl 2、Bax基因蛋白的表达及其与凋亡的关系。　方法　用胎龄 17～ 18dWistar大鼠的大脑皮层神经细胞进行原代分离培养。采用TUNEL染色方法、流式细胞术确立缺氧复氧神经细胞凋亡病理模型 ,并用免疫组织化学方法显示缺氧复氧不同时间Bcl 2、Bax基因的动态表达。　结果　 1.缺氧复氧可以使大鼠大脑皮层神经细胞发生凋亡 ,在分别缺氧 2、4、6、8h ,复氧 0h和18h组中 ,缺氧 4h开始出现凋亡细胞 ,随缺氧时间的延长 ,凋亡细胞数渐多 ,至缺氧 8h ,复氧 18h达高峰 ;2 .在单纯缺氧组中随缺氧时间延长 ,Bcl 2基因表达逐渐下降 ,Bax基因表达逐渐升高 ,两者呈显著负相关 ,而凋亡的发生也与Bax基因、Bcl 2基因表达呈显著正、负相关性。在缺氧复氧组中 ,未发现相关性。　结论　缺氧复氧可致胎鼠大脑皮层神经细胞发生凋亡 ;缺氧复氧所引起Bcl 2基因表达增高 ,Bax基因表达降低 。Objective The study is to investigate the dynamic expression of Bcl 2 and Bax protein in the apoptosis of primary cultured rat cortical neurons following hypoxia/reoxygenation(H/R)and its relation with apoptosis. Methods The cortical neurons of E16 17 days fetal rat was primarily cultured.The apoptosis model of primary cultured cortical neurons following H/R was established by using TUNEL staining and flow cytometry.The dynamic expression of Bcl 2 and Bax protein at different H/R time was investigated with immunohistochemical method. Results 1.H/R can cause apoptosis of primary cultured rat cortical neurons.In the experiment of H 2R 0,H 4R 0,H 6R 0,H\-8R\-0 and H 2R 18 ,H 4R 18 ,H 6R 18 ,H 8R 18 ,the apoptosis cells occurred after 4 hour hypoxia.The apoptosis cell increased with the time,and reaching peak value at H 8H 18 .2.In the experiment of solo hypoxia group,the expression of Bcl 2 protein decreased with hypoxia time,but Bax increased.They showed obviously negative correlationship.Furthermore,the rate of apoptotic cell also showed obviously negative correlation with Bcl 2 and positive correlation with Bax.However,in hypoxia/reoxygenation group,no correlation was found. Conclusion The apoptosis of primary cultured rat cortical neurons could be induced by H/R.That hypoxia causes the increase of the expression of Bcl 2 protein and the decrease of the expression of Bax protein could be one of the mechanisms of apoptosis.

关 键 词：原代培养 神经元 细胞凋亡 BCL-2 BAX 缺氧 大鼠 脑缺血 

分 类 号：R743.31[医药卫生—神经病学与精神病学]