摘要
目的探索能够应用于临床辨别早期缺血心肌生物学活性的敏感代谢变化指标。方法SD大鼠行冠状动脉前降支结扎术建立心肌缺血模型,分别结扎0(对照组)、5、20、45min。采用luciferin/luciferase法,RT-PCR法和免疫组织化学法分析心肌组织在不同缺血时间段梗死区、梗死边缘区及正常区ATP含量变化,葡萄糖调节蛋白75(grp75)和缺氧诱导因子1α(HIF1α)基因表达变化,以及细胞色素C释放、聚腺苷二磷酸核糖聚合酶(PARP)降解的情况。结果大鼠冠状动脉前降支分别结扎5、20、45min时,梗死区及梗死区边缘心ATP含量上升,并于20、45min时明显高于正常水平;grp75及HIF1α基因转录水平未见明显改变。免疫组化结果显示少数细胞细胞色素C的释放出现于冠状动脉前降支结扎5min,而PARP的降解发生较迟,出现于冠状动脉前降支结扎20min。冠状动脉前降支结扎45min时细胞色素C的释放和PARP的降解明显增强免疫反应呈现片状染色。结论大鼠急性心肌缺血后早期梗死区发生细胞凋亡,细胞色素C释放、PARP降解出现较早,可用作为临床辨别缺血心肌生物学活性的早期变化指标。
Purpose To find a kind of sensitive ways to check the earlier changes of ischemia myocardial biological activity that can be applied in clinic. Methods Induced myocardial infarction by ligation of the left anterior descending artery, the adult SD rats suffered from experimental myocardial infarction （MI） were divided into four groups： 0 （control）, 5, 20 and 45 min. The alteration of ATP content, the changes of expression of glucose regulate protein 75 （grp 75） gene and hypoxia-inducible factor 1α （HIF1α） gene, the release of cytochrome C and the degradation of Poly （ADP-ribose） polymerase （PARP） were detected in the infarction area, the margin of the infarction area and the ＂normal＂ area of the myocardial tissue respectively. Results （1） The content of the ATP in the infarc- tion area and the margin of the infarction area increased and was remarkably higher than the control in 20 and 45 min groups. （2） The changes of the expression of grp 75 gene and HIFla gene were not clear. （3） The immunohistochemistry showed that the release of cytochrome C began within 5 min in single cells of the infarction area after ligation and extended to large areas within 45 min. While the degradation of PARP began later than the release of cytochrome C, which began within 20 min MI and also extended to large areas within 45 min. Conclusions The release of cytochrome C and the degra-dation of PARP in infarction area occur early in the process of the apoptosis after acute MI and can be a sensitive biomarker for early acute MI diagnosis, treatment and prognosis.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2007年第4期477-481,共5页
Fudan University Journal of Medical Sciences
基金
上海市科学技术委员会科研计划项目课题(04JC14027)
