神经元缺血后caspase-3 mRNA表达的变化

The Changes of Caspase-3 mRNA Expression in Ischemic Neurons

目的 :明确caspase - 3是否脑缺血及缺血再灌注损伤基因治疗的适宜目的基因 .方法 :借助斑点杂交、原位杂交和核糖核酸印迹三种核酸杂交方法 ,用对环境无放射性污染的地高辛精标记探针 ,以PC12细胞和大鼠脑组织为研究对象 ,检测细胞和组织缺血缺氧前后caspase - 3基因转录水平的变化 .结果 :缺血缺氧及再灌注后caspase - 3mRNA明显高表达 ,而对照组表达很少 :用凝胶成像分析系统分析斑点杂交显色图像 ,对照组显色强度平均 9～ 32 ,而缺血缺氧组平均 78～ 2 18之间 .表明缺血缺氧及缺血再灌注能有效激活caspase - 3基因促进凋亡的作用 .提示 :caspase -

In order to clarify whether gene caspase-3 is the appropriate target gene for treating cerebral ischemia and reperfusion, we research the changes of it's transcription in pheochromocytoma PC12 cells and ischemic tissue of rat with the methods of dot blot, in situ hybridization and Northern blotting. The probe was labeled with nonradiated digoxigenin. The results showed that all ischemic groups remarkably up expressed caspase-3 mRNA after ischemia and reperfusion, while expression in control was very poor: testing the dots of dot blot with the gel image formation analysis system, found that the value of gray in the controls is from 9 to 32 yet 78～218 in ischemic groups. Suggest that ischemia, anoxia and reperfusion can activate the apoptosis induced by caspase-3. The conclusion is that caspase-3 is the proper targets of anti-sense RNA gene therapy of rat cerebral ischemia.