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热休克蛋白70(HSP70)在细胞修复、存活和维持细胞正常功能方面有着重要作用。作为分子伴侣,它起着心肌保护的作用。已经对重症心脏病人的心肌组织进行了蛋白组学研究,得到了HSP70在心衰病人心肌组织中较正常人心肌组织表达升高的结论,并且在血液中得到了进一步的验证。在进一步的离体细胞实验中用不同剂量的肿瘤坏死因子-alpha(TNF-α)刺激乳鼠心肌细胞,以观察不同时间点HSP70的动态表达情况。培养乳鼠心肌细胞,分别对细胞进行热休克(42℃)、TNF-α和缺血缺氧处理,在不同的时间点收获细胞,以观察HSP70的动态表达情况。用免疫化学、ELISA以及Western blotting的方法对HSP70蛋白进行分析。结果表明,在正常对照细胞中基本没有阳性信号出现,而在经缺血缺氧、热休克(42℃)以及TNF-α处理的细胞中有明显的阳性表达。以上研究首次在乳鼠心肌细胞中证明TNF-α诱导的HSP70表达具有时间和浓度依赖性。通过运用TNF-α对HSP70蛋白表达影响的研究,初步推断HSP70的表达模式,为体内诱导产生HSP70从而发挥心肌保护作用的研究提供一定的理论基础。
Heat shock protein 70 (HSP70) plays an important role in cell repair, survival and maintenance of normal cellular functions. As a molecular chaperone, it plays a role in myocardial protection. Proteomics has been carried out on myocardial tissue of patients with severe heart disease, and HSP70 expression in myocardial tissue of normal people with heart failure is higher than that in normal people. The result is further verified in the blood. In a further in vitro experiment, different doses of tumor necrosis factor-alpha (TNF-α) were used to stimulate neonatal rat cardiomyocytes to observe the dynamic expression of HSP70 at different time points. The cultured neonatal rat cardiomyocytes were treated with heat shock (42 ℃), TNF-α and hypoxia-ischemia, respectively. The cells were harvested at different time points to observe the dynamic expression of HSP70. HSP70 protein was analyzed by immunochemistry, ELISA and Western blotting. The results showed that there was almost no positive signal in normal control cells, but significant positive expression in cells treated with hypoxia and hypoxia, heat shock (42 ℃) and TNF-α. The above study for the first time in neonatal rat cardiomyocytes that TNF-αinduced HSP70 expression in a time and concentration-dependent manner. Through the study of the effect of TNF-α on the expression of HSP70 protein, the expression pattern of HSP70 was preliminarily deduced, which provided a theoretical basis for the study of the protective effect of HSP70 induced in vivo.