PI3KAkt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.docVIP

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 重庆，重庆医科大学附属第一医院胸心外科) [摘要]目的 探讨硫化氢后处理对原代培养的新生大鼠心肌细胞缺氧复氧损伤的保护作用及可能机制。方法 以硫氢化钠(NaHS)作为外源性硫化氢供体。原代培养新生大鼠的心肌细胞随机分为5组：(CON组)，②缺氧-复氧组(Hypoxia-reoxygenation，HR组)，③二甲基亚砜(Dimethyl sulfoxide，DMSO组)，④硫化氢后处理组(Hydrogen sulfide，H2S组)，⑤LY294002+H2S后处理组(LY+H2S组)(n=6)。分别于缺氧前和复氧2 h检测各组的心肌细胞存活率、培养液中CK和LDH的活性；复氧末，用流式细胞学技术检测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 通过比较缺氧前与复氧2h差值可知，复氧2h时，H2S组心肌细胞存活率较HR组升高显著[(27.37±2.11)vs(36.42±2.3)，P＜0.01]，CK、LDH活性以及心肌细胞凋亡率明显降低[(98.45±24.59)vs(193.55±46.98)，(144.52±20.10)vs(201.66±21.37)，(10.37±1.51)vs(17.38±2.04)，均为P＜0.01]；同时，HIF-1α和p-Akt蛋白表达水平明显升高[(63.93±4.64)vs(41.82±6.14)，(62.59±2.11)vs(41.04±4.01)，均为P＜0.01]。然而，在H2S后处理同时加入PI3K/Akt信号通路特异性抑制剂LY294002后，与H2S组相比，心肌细胞存活率降低明显(34.45±3.34)，CK、LDH活性以及心肌细胞凋亡率升高显著[(173.69±30.34，(197.38±18.21)，(15.17±1.56)均为P＜0.01]，并且，HIF-1α和p-Akt蛋白表达水平亦降低明显[(51.79±4.27)，(47.42±3.15)，分别为P＜0.05和P＜0.01]。结论 H2S后处理可通过PI3K/Akt信号通路促进HIF-1α表达，从而拮抗心肌细胞的缺氧复氧损伤。 [关键词]硫化氢；心肌缺血再灌注损伤；缺氧诱导因子1α；凋亡；PI3K/Akt Hydrogen sulfide postconditioning protects cardiomyocytes against hypoxia - reoxygenation injury via PI3K/Akt pathway Li Gang，Yang Shuangqiang(Department of Cardiothoracic Surgery，the First Affiliated Hospital，Chongqing Medical University，Chongqing400016 China) [Abstract] Objective To investigate the protective effect and possible mechanism of hydrogen sulfide postconditioning on primary cultured cardiomyocytes of neonatal rats injured by hypoxia-reoxygenation. Methods Sodium hydrosulfide was used as an exogenous donor of hydrogen sulfide. Primary cultured cardiomyocytes of neonatal rats were randomly divided into five groups：(1)control group，2) hypoxia-reoxygenation group，3)dimethyl sulfoxide group，4)hydrogen sulfide postconditioning group，5)LY294002 + hydrogen sulfide postconditioning group(n=6). Cardiomyocytes viability and creatine kinase，lactate dehydrogenase in the medium of cultured cardiomyocytes were measured respectively in each group at the point of before hypoxia， hypoxia an