姜黄素对缺血-再灌注H9c2心肌细胞的保护作用及对热休克蛋白27表达的影响

Curcumin protects H9c2 myocytes from ischemia-reperfusion injury by inducing expression of Hsp27 protein

目的探讨姜黄素对缺血一再灌注损伤的H9c2心肌细胞热休克蛋白27（Hsp27）表达的影响及意义。方法利用缺血台氏液对体外培养的H9c2心肌细胞模拟缺血一再灌注处理．同时随机分为缺血一再灌注、缺血一再灌注＋姜黄素组和对照组，采用流式细胞术检测各组细胞凋亡率，JC-1法检测各组线粒体膜电位情况．噻唑蓝法检测各组细胞存活率，Westernblot法检测各组Hsp27蛋白表达水平。结果姜黄素组干预后的早期、晚期和总细胞凋亡率[分别为（4．94±0．6）％、（5．8．4±1．3）％和（10．7±1．5）％]均较缺血一再灌注组[分别为（8．6±2．1）％、（19．3士2．4）％和（27．9±3．2）％]明显降低．差异均有统计学意义（P〈0．05或001）。姜黄素组低电位细胞比例数[（38．2±2．4）％]较缺血一再灌注组[（59．3±7．5％）％]明显降低，而姜黄素组细胞的后续存活率和Hsp27蛋白表达水平[分别为（813±13）％和（109．1±9．3）％]均显著高于缺血一再灌注组[分别为（30．5±2．0）％]和（78．6土6．6）％]，差异均有统计学意义（均P〈0．01）。结论诱导Hsp27蛋白高表达可能是姜黄素减轻心肌细胞缺血一再灌注损伤的重要机制之一。

Objective TO investigate the effects of curcumin on cellular injury and heat shock protein 27 （Hsp27） expres- sion in H9c2 myocytes subjected to simulated ischemia- reperfusion （IR）. Methods Cultured H9c2 cells were randomly divided into 3 groups： normal control group, IR group and IR＋curcumin group. The procedure of IR was performed in H9c2 cells by ex- posing to a simulated ischemia solution for 90 min followed by 30 min of reperfusion with the normal Tyrode solution. Then cell apoptosis, mitochondrial membrane potential （MMP）, and cell viability were detected by flow cytometry, JC-1 assay, and methylthiazoletetrazolium assay, respectively. Hsp27 protein expression was evaluated by Western blot analysis. Results Com-pared with IR group, the early, late and total cell apoptosis rate in IR＋curcumin group were significantly decreased, respectively [ （4.9 ± 0.6） % vs （8.6 ± 2.1） %, （5.8 ± 1.3） % vs （19.3 ± 2.4）%, （10.7 ± 1.5） % vs （27.9 ± 3.2） %, all P〈O.05 or 0.01]. The proportion of cells with low MMP in IR＋curcumin group was lower than that in IR group [（38.2 ± 2.4） % vs （59.3 ± 7.5%） %, P〈O.01]. Compared with IR group, cell survival after IR and expression level of Hsp27 protein in IR＋curcumin group were significantly increased, re- spectively [（81.3 ± 1.3） % vs （30.5 ± 2.0）%, （109.1 ± 9.3）% vs （78.6 ± 6.6） %, all P〈O.01]. Conclusion Curcumin may protect cardiac cells from ischemia-reperfusion injury by inducing expression of Hsp27 protein.