缺氧诱导因子-1α在热耐受对抗高温诱导的大鼠心肌细胞凋亡中的作用  

Effect of hypoxia inducible factor-1α on thermotolerance against hyperthemia induced cardiomyocytes apoptosis

作　　者：黄碧君[1] 程晓曙[1] 

机构地区：[1]南昌大学第二附属医院心血管内科江西省分子医学重点实验室,330006

出　　处：《中华心血管病杂志》2013年第9期785-789,共5页Chinese Journal of Cardiology

基　　金：“十一五”国家科技支撑计划资助项目（2008BA168B00）

摘　　要：目的 研究缺氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）在非致死性高温诱导的热耐受过程中的表达情况及其在心肌热耐受保护中的作用。 方法 培养H9c2心肌细胞，HIF-1α抑制剂YC-1干预24 h后进行热耐受（40 ℃，3 h）或高温（43 ℃，2 h）处理。实验分为8组，每组实验重复3次。分别为常温对照组（对照组），37 ℃ ；热耐受组（T组）；热耐受/高温组（T/H组）；高温组（H组）；DMSO＋常温对照组（DMSO组）；YC-1干预＋热耐受组（YC-1＋T组）；YC-1干预＋热耐受/高温组（YC-1＋T/H组）；YC-1干预＋高温组（YC-1＋H组）。流式细胞仪检测细胞凋亡率。Western blot检测HIF-1α和半胱氨酸天冬氨酸蛋白酶（caspase-3）的表达水平。 结果 T组细胞凋亡率与对照组比较差异无统计学意义；H组细胞凋亡率［（17.35±1.07）%］显著高于对照组［（7.52±1.55）%］, P〈0.01; T/H组细胞凋亡率［（12.58±1.97）%］显著低于H组［（17.35±1.07）%］,P〈0.01。YC-1＋T组细胞凋亡率与DMSO组比较差异无统计学意义；YC-1＋T/H组细胞凋亡率与YC-1＋H组比较差异无统计学意义。YC-1＋T/H组凋亡率［（23.75±1.92）%］显著高于T/H组［（12.58±1.97）%］，P〈0.01；YC-1＋H组细胞凋亡率［（24.89±1.83）%］显著高于H组［（17.35±1.07）%］，P〈0.01。T组HIF-1α蛋白表达水平显著高于对照组（P〈0.05）；T/H组HIF-1α蛋白表达水平显著高于H组（P〈0.01）。YC-1＋T组、YC-1＋T/H组及YC-1＋H组HIF-1α蛋白表达水平均显著低于DMSO组（P均〈0.01）。T组caspase-3蛋白表达水平与对照组比较差异无统计学意义；T/H组caspase-3蛋白表达水平显著低于H组（P〈0.01）。YC-1＋T组、YC-1＋T/H组及YC-1＋H组caspase-3蛋白表达水平均显著高于DMSO组（P均〈0.01）。YC-1＋T组caspase-3蛋白表达水平与DMSO组比较差异无统计学意义；YC-1＋T/H组caspase-3蛋白表达水�Objective To investigate the expression changes and effects of hypoxia inducible factor- 1α （HIF-1α） on non-lethal high temperature induced tbermotolerance and its role on thermotolerance protection. Methods H9c2 cardiomyocytes were cultured and pretreated with the HIF-1α inhibitor YC-1, the cells were then subjected to normal temperature （37 ℃ ） , tbermotolerance induction （40 ℃, 3 h） , or hyperthermia （43 ℃, 2 h）. The cells were divided into 8 groups （n =3 each） ： normal temperature control group ; thermotolerance group ; thermotolerance/hypertherrnia group ; byperthermia group ; DMSO ＋ normal temperature group; YC-1 ＋ thermotolerance group; YC-1 ＋ thermotolerance/hyperthermia group; YC-1 ＋ hyperthermia group. Cell apoptotie rate was assessed by flow cytometry. Western blot was used to detect the expression of HIF-1α and easpase-3. Results Flow cytometry results showed that apoptosis rate was similar between eontrol group and thermotolerance group, between DMSO ＋ normal temperature group and YC-1 ＋ thermotolerance group, between YC-1 ＋ thermotolerance/hyperthermia group and YC-1 ＋ hyperthermia group, but was significantly higher in hyperthermia group [ （17.35 ± 1.07）% ] than in control group [（7.52 ± 1.55 ）%, P 〈 0.01 ] which was partly reduced in thermotolerance/hyperthermia group [ （ 12. 58 ± 1.97） %, P 〈 0. 01 vs. thermotoleranee group]. Cell apoptosis rate of YC-1 ＋ thermotolerance/ hyperthermia group （23.75± 1.92） % was significantly higher than that of thermotolerance/hyperthermia group [（12.58±1.97）%, P〈0.01], and in YC-1 ＋hyperthermia group [（24.89 ±1.83）%] than in hyperthermia group [ （17.35 ±1.07）%, P 〈 0. 01 ]. HIF-1α expression was obviously upregulated in thermotolerance cells compared with control cells, in thermotolerance/hyperthermia cells than in hyperthermia cells, in YC-1 ＋ thermotolerance group, YC-1 ＋ thermotoleranee/hyperthermia group and YC-1 ＋ hyperthermia group than in DMSO group （ al

关 键 词：肌细胞 心脏 缺氧诱导因子1 α亚型 细胞凋亡 热耐受 

分 类 号：R363[医药卫生—病理学]