热休克蛋白90在缺氧心肌细胞中表达的初步研究

The research of heat shock protein 90's expression in hypoxic rat myocardial cells

观察热休克蛋白90(HSP90)在缺氧心肌细胞中的表达变化规律,并初步阐明HSP90在早期心肌缺氧损害中的作用。方法:原代培养SD大鼠乳鼠心肌细胞,随机分为正常对照组、缺氧组和加HSP90阻断剂格尔霉素(geldanamycin,GA)预处理后再缺氧组(GA+缺氧组),蛋白免疫印迹法(western blot)及间接免疫荧光法检测缺氧1、3、6、12、24h后心肌细胞中HSP90蛋白分布表达情况,并检测细胞上清中肌酸激酶同工酶(CK-MB)及乳酸脱氢酶(LDH)的含量变化。结果:心肌细胞缺氧3h后HSP90蛋白在胞浆内表达量升高,持续到12h达到峰值。与单纯缺氧组相比,GA+缺氧组中LDH,CK-MB含量在不同时相点均有显著升高。结论:缺氧早期即可引起心肌细胞胞浆中HSP90蛋白表达增强,HSP90可能在早期心肌缺氧损害中发挥其内源性抗损伤机制。

Objective： the main aim of the present work is to observe the expression of heat shock protein 90 （HSP90） in hypoxk rat myocardial cells. Also elucidated is the effect of HSP90 on the early hypoxia induced cardiac musde injury. Methods ： SD fetal rat myocardial cells were cultured in vitro, stochastically divided into three groups： normal control, hypoxia group and hypoxia group pretreated by geldanamycin （GA-added hypoxia group）. Samples of myocardial cells were taken for the determination of HSP90 expression by Western blot analysis and indirect immunofluorescence at 1, 3, 6, 12, 24h after hypoxia. Changes of CK-MB and LDH content were detected. Results：expression level of HSP90 increased in rat myocardial cells endochylema after 3h of hypoxic injury, and the level reached highest at12h after hypoxia. Compared to the hypoxia group, the level of LDH, CK-MB in GA-added hypoxia group is significantly higher at every point. Conclusion：hypoxia injury causes an increased expression of HSP90 in rat myocardial cells, and HSP90 may have endogenous anti-injury effect on early hypoxia injury.