体外心肌细胞缺血再灌注损伤模型的建立  

Establishment of Ischemic/Reperfusion Injury Model of Myocardial Cell in vitro

作　　者：张娜娟[1] 张彤彤[1] 刘志祥[1] 裴斐[1] 魏华英[1] 马刚[1] 杨超[1] 杨春蕾[1] 

机构地区：[1]四川大学生命科学学院医学细胞生物学教研室,成都610064

出　　处：《四川动物》2009年第3期440-443,共4页Sichuan Journal of Zoology

摘　　要：目的建立体外心肌细胞缺血再灌注(I/R)模型,为研究各种心血管疾病及药物治疗提供基础。方法体外培养新生SD乳鼠心肌细胞,实验分正常组和缺血再灌注组。倒置显微镜及透射电子显微镜下观察心肌细胞缺血再灌注前后的形态变化,MTT、流式细胞术检测细胞凋亡情况,免疫组化和Western Blot检测Bcl-2/Bax基因表达。结果I/R前后心肌细胞形态发生明显变化。与正常组相比,I/R组心肌细胞凋亡率明显增加,细胞活性降低。免疫组化和Western Blot检测结果均出现I/R后Bcl-2蛋白表达下调,Bax蛋白表达上调,Bcl-2/Bax显著减小的现象。结论体外培养的心肌细胞I/R损伤各项指标检测与动物体内实验结果相吻合,体外心肌细胞缺血再灌注模型建立成功。Objective To establish an ischemic/reperfusion （I/R） model of myocardial ceils in vitro. Methods Neonatal myocardial cells from SD rats were cultured in vitro, and they were divided into a normal group and I/R group. Morphological changes before and after I/R were observed by inverted microscopy and transmission electron microscopy. Apoptosis was detected by MTT assay and flow cytometry （FCM）. Expressions of Bcl-2/Bax proteins were measured by immuno- histochemistry and Western blot. Results There were great changes in the morphology of myocardial cells before and after I/R. An increase of the apoptotic rate and a decreased cell activity were observed as compared with those in the control group. Both immunohistochemistry and Western blot showed a down regulation in the expression of Bcl-2 protein and an upregulated expression of Bax protein. The rate of Bcl-2/Bax was obviously decreased. Conclusion Apoptotic indices of myocardial cell cultured in vitro induced by I/R are in assistant with those in vivo, so an I/R model of myocardial cell in vitro was successfully established.

关 键 词：心肌细胞 缺血再灌注 BCL-2/BAX 凋亡 

分 类 号：R-332[医药卫生]