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目的:观察富含氢气(H_2)的培养基对血清-葡萄糖剥夺(SGD)致心肌细胞损伤的保护效应,探讨血红素加氧酶1(HO-1)在其中的作用。方法:用SGD的方法处理H9c2心肌细胞,建立缺血性心肌病的细胞模型。在SGD处理后,分别给予正常培养基和富含H_2的培养基以观察H_2的心肌细胞保护作用,给予选择性HO-1抑制剂锌原卟啉IX(ZnPP IX)以抑制HO-1的作用;各处理因素完成后,检测细胞存活率、羟基自由基(OH·)的含量及HO-1蛋白的表达。结果:SGD处理6~30 h,时间依赖性地降低H9c2心肌细胞的存活率,相关性分析显示r为-0.94。SGD处理6 h后再正常培养24 h可明显增加细胞内OH·的含量,并上调HO-1蛋白的表达,与对照组比较,均P<0.01。ZnPP IX通过抑制HO-1的作用能明显加重SGD处理诱导的心肌细胞存活率降低(P<0.05)。在SGD处理后给予富含H_2的培养基能减轻SGD引起的细胞损伤,使细胞存活率明显提高(P<0.05),胞内OH·的含量显著降低(P<0.01)。富含H_2的培养基还能使SGD诱导的HO-1表达进一步上调,而ZnPPⅨ部分取消了H_2诱导的细胞保护作用(P<0.05)。结论:富含H_2的培养基可减轻血清-葡萄糖剥夺引起的心肌细胞损伤,其机制不仅与直接清除OH·有关而且还与上调抗氧化酶系统HO-1的表达有关。
Objective: To observe the protective effect of hydrogen-rich medium on cardiomyocyte injury induced by serum-glucose deprivation (SGD), and to explore the role of heme oxygenase-1 (HO-1) in it. Methods: H9c2 cardiomyocytes were treated with SGD to establish a cell model of ischemic cardiomyopathy. After SGD treatment, normal and H 2 -rich media were respectively administrated to observe the cardiomyocyte protective effect of H 2, and the selective HO-1 inhibitor zinc protoporphyrin IX (ZnPP IX) was administered to inhibit the effect of HO-1 After treatment, the cell viability, the content of hydroxyl radical (OH ·) and the expression of HO-1 protein were detected. Results: After treated with SGD for 6 ~ 30 h, the survival rate of H9c2 cardiomyocytes was decreased in a time-dependent manner. Correlation analysis showed that r was -0.94. After cultured for 6 h, SGD normal cultured for 24 h significantly increased the content of OH · and increased the expression of HO-1 protein, both P <0.01 compared with the control group. ZnPP IX significantly increased the decrease of survival rate of cardiomyocytes induced by SGD treatment by inhibiting the effect of HO-1 (P <0.05). The treatment with SGD enriched H 2 could reduce the cell injury caused by SGD, significantly improve the cell survival rate (P <0.05), and significantly decrease the intracellular OH · (P <0.01). H 2 -rich medium also induced a further up-regulation of HO-1 expression induced by SGD, whereas ZnPPⅨ partially abolished H 2 -induced cytoprotection (P <0.05). CONCLUSION: H 2 -rich medium can reduce the damage of cardiomyocytes induced by serum-glucose deprivation, which is not only related to the direct scavenging of OH · but also the up-regulation of the expression of HO-1, an antioxidant enzyme system.