Analysis of apoptosis signal and stem cells in congenital heart disease model rats.

本实验制作了缺氧和容量超负荷的大鼠先天性心脏病模型，并观察了心肌组织形态学和生化指标的变化。作为该方法，收集每次负荷结束后的心肌组织，并且1）TUNEL染色以评估心肌细胞凋亡，2）分析细胞凋亡的信号通路，进行FasL、casoase 9和ASK 1的心肌的Western印迹，以及3）分析FasL、casoase 9和ASK 1的上游信号，实施作为内质网应激的标志物的CHOP的RT-PCR。结果表明：1）缺氧模型大鼠TUNEL阳性率明显高于假手术组; 2)心肌组织中FasL蛋白表达在缺氧组、容量超负荷组和假手术组中无明显变化。而缺氧组caspase 9和ASK 1表达升高。3)RT-PCR分析CHOP，缺氧组高于其他各组。了解内质网应激参与了缺氧负荷，并对ASK 1上游的未来信号进行分析。通过基因芯片的基因检索，分别找到在缺氧组和容量超负荷组中表达增强的基因，即利用缺氧和容量超负荷心肌组织来检索心肌细胞凋亡相关基因的变化。

A hypoxia and a volume overload rat model of congenital heart disease are made, and the morphological change of the cardiac muscle tissue and the biochemical changes have been examined. As the method, the cardiac muscle tissue after each load ended was gathered, and 1)TUNEL dye to evaluate cardiac muscle apoptosis, 2) to analyze signal pathway of apoptosis, westernblot of the cardiac muscle of FasL,casoase9, and ASK1 was done, and 3) to analyze the upstream signal of FasL, casoase9, and ASK1,RT-PCR of CHOP that is the marker of the endoplasmic reticulum stress was enforced. Result is that1) hypoxia model rats were higher in TUNEL positive than sham rats. 2) in westernblot analysis incardiomyosite, no remarkable change was observed in FasL among hypoxia group and volume overload group, sham group. But caspase9 and ASK1 were higher in hypoxia group. 3) CHOP analysis of RT-PCR, hypoxia group was higher than other groups. It was understood that the endoplasmic reticulum stress took part about a hypoxia loading, and the future signal of the upstream of ASK1 willbe analysed. The gene that was accentuated in the hypoxia and the volume overload group respectively was found though the gene retrieval by DNA microarray, that was enforced by using thehypoxia and the volume overload cardiac muscle organization to retrieve the change of the gene related to cardiac muscle apoptosis.