硫化氢后处理激活缺氧诱导因子1α保护缺血心肌 - 第三军医大学学报.docVIP

硫化氢后处理激活缺氧诱导因子1α保护心肌细胞缺氧复氧损伤 PI3K/Akt通路在硫化氢后处理保护心肌细胞缺氧-复氧损伤中的作用 （400016 ） （NaHS）分为5组：（CON）-复氧组（HR）（DMSO）H2S后处理组（H2S）LY294002+H2S后处理组（LY+H2S）2 h和复氧2 h检测各组的心肌细胞存活率、培养液中CK和LDH的活性；流式细胞学技术检测复氧末各组心肌细胞凋亡情况；Western blot检测HIF-1α、p-Akt与Akt蛋白的表达情况。结果 缺氧2 h，2-5组上述指标较CON组显著？（P0.01）2 h时，H2S组心肌细胞存活率较HR组升高显著（P0.01）CK与LDH活性明显降低（P0.01）HIF-1α和p-Akt蛋白表达水平明显升高（P0.01）PI3K/Akt信号通路特异性抑制剂LY294002后逆转了硫化氢后处理的上述效应。结论 H2S后处理可通过PI3K/Akt信号通路激活HIF-1α拮抗心肌细胞的缺氧复氧损伤。 [关键词]硫化氢；心肌缺氧复氧损伤；缺氧诱导因子1α；凋亡；PI3K/Akt Hydrogen sulfide postconditioning protects cardiomyocytes against MIRI via the activation of HIF-1α Li Gang，Yang Shuangqiang（Department of Cardiothoracic Surgery，the First Affiliated Hospital，Chongqing Medical University，Chongqing400016 China） [Abstract] Objective To investigate the effects and possible mechanism of hydrogen sulfide （H2S） postconditioning on primary cultured cardiomyocytes of neonatal rats injured by hypoxia-reoxygenation. Methods NaHS was used as an exogenous donor of H2S. Primary cultured cardiomyocytes of neonatal rats were randomly divided into five groups. （1）control group （CON group）， （2） hypoxia-reoxygenation group （HR group）， （3） menstruum control group （DMSO group）， （4） H2S postconditioning group （H2S group）， （5） LY294002+H2S postconditioning group （LY+H2S group）. Cardiomyocytes viability and creatine kinase （CK）， lactate dehydrogenase （LDH） in the medium of cultured cardiomyocytes were measured respectively in each group at the point of before hypoxia， hypoxia and reoxygenation 2h. Apoptosis of cardiomyocytes were detected by flow cytometry and the expression of HIF-1α，p-Akt and Akt protein were tested by Western Blot. Results Cardiomyocytes viability and the activity of CK and LDH had no significant difference in each group before hypoxia. At the point of hypoxia 2h， the above data of last four groups had remarkable difference compared with CON group （all P0.01）. But， we failed to detect any significant between-group differences among the last four groups. At the point