实验性脑出血后的细胞凋亡和凋亡相关基因转录、表达水平  

Apoptosis and its related gene transcription and expression following experimental intracerebral hemorrhage

作　　者：张尉华[1] 饶明俐[1] 吴江[1] 胡利华[1] 张颖[1] 

机构地区：[1]吉林大学第一医院神经内科,吉林长春130021

出　　处：《中风与神经疾病杂志》2005年第1期10-12,共3页Journal of Apoplexy and Nervous Diseases

基　　金：卫生部临床学科重点项目 (2 0 0 13 14 4) ;吉林省科技发展计划项目 (2 0 0 3 0 5 3 6-1)

摘　　要：目的　通过实验性脑出血动物模型研究脑出血后的细胞凋亡和凋亡相关基因转录、表达水平。方法　自体动脉血注入法建立实验性脑出血动物模型 ,分别行 Bcl- 2、bax、caspase- 3和 TNF- α免疫组化染色 ,细胞凋亡检测采用 TUNEL 法、流式细胞仪检测及 DNA电泳 ,RT- PCR检测血肿周围组织 caspase- 3m RNA的含量。结果　血肿形成后 6 h～ 7d,周围组织中 bcl- 2阳性细胞数逐渐下降至 1d达到最低 ;bax、caspase- 3、TNF- α以及 TU NEL 阳性细胞数逐渐上升 ,分别至 1～ 2 d、2～ 3d、3d和 3～ 5 d达到峰值。血肿周围组织细胞周期检测均可见 G1峰前形成 Ap亚峰 ,Ap亚峰细胞数逐渐增加 ,在 5 d形成高峰。2～ 7d组可见断裂 DNA片段形成的梯形条带。Caspase- 3m RNA的相对表达量逐渐增加 ,2 d达到峰值。结论　实验性脑出血后血肿周围组织中存在细胞凋亡 ,并有凋亡相关基因的表达 ,凋亡细胞数逐渐增加 ,5 d达峰值。Objective The apoptosis and its related gene transcription and expression following experimental intracerebral hemorrhage(ICH) were studied by rat ICH model. Methods The rat model of experimental ICH was established by injection of arterial blood into the caudate nucleus. Immunohistochemical(IHC) stains of bcl-2,bax,caspase-3 and TNF-α,TUNEL,flow cytometry and DNA electrophoresis were studied. The relative content of caspase-3 mRNA around the hematoma was detected by method of RT-PCR. Results After the formation of hematoma,the number of positive cells of bcl-2 IHC stain gradually decreased and arrived at the valley at the time of 1d,while that of bax,caspase-3,TNF-α and TUNEL gradually increased and peaked at the 1～2d、2d～3d、3rd and 3d～5d respectively. The Ap (Apoptosis) subpeak could be observed in each group with the method of flow cytometry. The percents of cellular number in the Ap subpeak gradually increased and peaked at the 5th. The ladder-like stripe formed by rupture of DNA might be observed in the group of the time points of 2d～7d with the method of DNA electrophoresis.. The content of caspase-3 mRNA gradually increased and peaked at 2d. Conclusion Cellular apoptosis and its related gene transcription and expression could be observed following experimental intracerebral hemorrhage. The quantity of apoptosis cells gradually increased and peaked at the 5th.

关 键 词：实验性脑出血 凋亡相关基因 caspase-3 细胞凋亡 血肿周围组织 表达水平 TNF-α 转录 RNA DNA片段 

分 类 号：R743.34[医药卫生—神经病学与精神病学] R735[医药卫生—临床医学]