缺氧后适应与心肌营养素-1对心肌细胞缺氧复氧的保护作用及机制探讨  

Protective effects of hypoxic postconditioning and cardiotrophin-1 on cardiomyocytes after hypoxia-reoxygenation and the mechanism

作　　者：周贻军[1] 赵亚男[1] 赵鑫[1] 缪绯[1] 王先宝[1] 颜竞[1] 张秀丽[1] 刘映峰[1] 杨平珍[1] 

机构地区：[1]南方医科大学珠江医院,广州510280

出　　处：《山东医药》2013年第44期7-9,13,共4页Shandong Medical Journal

基　　金：广东省自然科学基金资助项目(10151051501000038);广东省广州市海珠区科普计划高血压基地构建项目[KP2010(T)-11)]

摘　　要：目的探讨缺血后适应与心肌营养素-1(CT-1)对心肌细胞缺氧复氧的保护作用及其机制。方法将体外培养的H9C2细胞株分为正常对照组(a组)、缺氧复氧组(b组)、缺氧复氧+缺氧后适应组(c组)、缺氧复氧+缺氧后适应+CT-1组(d组)、缺氧复氧+缺氧后适应+CT-1+ERK抑制剂组(e组)、缺氧复氧+缺氧后适应+CT-1+二甲基亚砜组(f组),MTS法检测各组细胞存活率、流式细胞仪检测细胞凋亡率、Western blot检测细胞外调节激酶(ERK1/2)和磷酸化ERK1/2(p-ERK1/2)的表达、Q-PCR检测Bad mRNA的表达。结果与a组比较,其他各组细胞存活率下降,细胞凋亡率、Bad mRNA和p-ERK1/2(除外e组)表达量增加,P均<0.05;与b组比较,c组细胞存活率、p-ERK1/2表达量均增加,Bad mRNA表达、凋亡率下降,P均<0.05;与c组比较,d组细胞存活率、pERK1/2表达量均增加,细胞Bad mRNA表达、凋亡率下降(P均<0.05);与d组比较,e组细胞存活率下降,细胞凋亡率、Bad mRNA表达量增加,p-ERK1/2表达量下降;d、e组各项指标指标比较差异无统计学意义。结论缺氧后适应可减少心肌细胞缺氧复氧损伤,联合CT-1后保护作用明显加强,而ERK1/2抑制剂可阻断这种保护作用;该保护机制可能与CT-1激活ERK1/2信号通路,下调Bad mRNA的表达有关。Objective To investigate the protective effects of hypoxic postconditioning and cardiotrophin-1 （CT-1） on cardiomyocytes after hypoxia-reoxygenation and the mechanism.Methods H9C2 cell lines were cultured in vitro,and then were divided into the normal control group （group a）,hypoxia-reoxygenation group （group b）,hypoxia-reoxygenation ＋ hypoxic postconditioning group （group c）,hypoxia-reoxygenation ＋ hypoxic postconditioning ＋ CT-1 group （group d）,hypoxia-reoxygenation ＋ hypoxic postconditioning ＋ CT-1 ＋ ERK1/2 inhibitor group （group e） and hypoxia-reoxygenation ＋ hypoxic postconditioning ＋ CT-1 ＋ DMSO group （group f）.MTS method was applied to detect cell survival rate,flow cytometry was used to detect the apoptosis rate,Western blot was employed to detect the p-ERK1/2 expression and Q-PCR was used to detect Bad mRNA expression.Results Compared with group a,the cell survival rate decreased,the apoptosis rate and Bad mRNA expression increased in other groups except group e （all P ＜ 0.05）.Compared with group b,the cell survival rate and p-ERK1/2 expression increased and Bad mRNA expression and apoptosis rate dropped in the group c （all P ＜0.05）.Compared with group c,the cell survival rate and p-ERK1/2 expression increased,Bad mRNA expression and apoptosis rate decreased in the group d （all P ＜0.05）.Compared with group d,the cell survival rate and the p-ERK1/2 expression decreased,the apoptosis rate and Bad mRNA expression increased in the group e.No significant difference was found between group d and e.Conclusions Hypoxic postconditioning can reduce the myocardial hypoxia-reoxygenation injury and the protective effect can be enhanced significantly by combining CT-1,but ERK1/2 inhibitor blocks the protective effect.The mechanism may be that ERK1/2 signaling pathway is activated,and then down-regulates Bad mRNA expression.

关 键 词：心肌缺血再灌注损伤 缺氧后适应 心肌营养素-1 细胞外信号调节激酶 BAD 

分 类 号：R542.2[医药卫生—心血管疾病]