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

建立先天性心脏病缺氧和容量过载大鼠模型，观察心肌组织形态学变化和生化变化。该方法收集每次负荷结束后的心肌组织，1)TUNEL染色评估心肌凋亡，2)分析凋亡信号通路，对心肌FasL、casoase9和ASK1进行westernblot， 3)分析FasL、casoase9和ASK1上游信号，对内质网应激标志物CHOP进行RT-PCR。结果：1)缺氧模型大鼠TUNEL阳性高于假手术大鼠。2)心肌组织westernblot分析显示，缺氧组、容量超负荷组、假手术组FasL无明显变化。低氧组caspase9、ASK1升高。3) RT-PCR CHOP分析，缺氧组高于其他组。据了解，内质网应激参与了缺氧负荷，ASK1上游的未来信号将被分析。通过DNA芯片基因检索，分别发现在缺氧组和容量超负荷组中被强化的基因，利用缺氧组和容量超负荷心肌组织来检索心肌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.