心肌营养素-1预处理减轻缺氧复氧对心肌细胞损伤的机制研究  

Protective effect of cardiotrophin-1 against hypoxia-reoxygenation injury in H9C2 cardiomyocytes

作　　者：李志聪[1] 游伟[1] 缪绯[1] 杨天潇 刘映峰[1] 

机构地区：[1]南方医科大学珠江医院心血管内科,广东广州510282

出　　处：《热带医学杂志》2014年第5期603-606,610,共5页Journal of Tropical Medicine

基　　金：广东省自然科学基金(10151051501000038);海珠区科普计划(KP2010T-11)

摘　　要：目的观察心肌营养素-1(CT-1)预处理对大鼠心肌细胞缺氧复氧损伤模型细胞损伤、凋亡及血红素加氧酶-1(HO-1)表达的影响,进一步探讨其心肌保护机制。方法选择H9C2大鼠心肌细胞株,实验分为对照组、缺氧复氧组(H2/R24组)、各浓度CT-1预处理组,酶标仪检测细胞上清中LDH释放率,RT-qPCR检测HO-1 mRNA,Western Bolt检测活Cleaved Caspase3及HO-1蛋白。设计合成靶向HO-1的特异性siRNA片段,通过脂质体转染至H9C2细胞中,48 h后给予CT-1预处理及缺氧复氧(si-HO-1组)并重复检查HO-1 mRNA及蛋白的表达、LDH释放率、Cleaved Caspase3表达验证干扰效果。结果与H2/R24组相比,各浓度CT-1预处理组细胞上清LDH释放率下降,Cleaved Caspase3表达减少,而HO-1 mRNA及蛋白表达水平与H2/R24相比均明显增加。经siRNA干扰后,si-HO-1组HO-1表达水平与对照组相当,较H2/R24组、CT-1组则明显减少;而LDH释放率、Cleaved Caspase3蛋白表达较H2/R24组下降,而较CT-1组升高。结论 CT-1能通过上调HO-1表达,减轻心肌细胞缺氧复氧损伤。Objective To investigate whether cardiotrophin-1 （CT-1） increases expression of heme oxygenase-1 （HO-1） in cardiomocytes, and whether CT-1-induced HO-1 protects against hypoxia/reoxygenation （H/R） jnjury. Methods H9C2 cardiomyocytes were exposed to H/R injury with or without preincubation with CT-1. The release rate of LDH in cells supernatant was measured. Cleaved caspase3 and HO-1 protein expressions were analyzed by Western blot. HO-1 mRNA expression was analyzed by RT-qPCR. HO-1-targeted siRNA was designed and synthesized, and then transfected into H9C2 cardiomyocytes via Lifpofectamine2000 mediation. The transfected H9C2 cardiomyocytes were exposed to H/R injury with preincubation with CT-1. Expressions of HO-1 mRNA and protein, release rate of LDH and expression of Cleaved caspase3 were analyzed again. Results Compared with hypoxia-reoxygenation group, the release rate of LDH and Cleaved caspase3 protein expression in CT-1 pretreatment groups were decreased significantly and the levels of HO-1 mRNA, and HO-1 protein were significantly increased. HO-I targeted siRNA could down-regulate the HO-1 expression of H9C2 cardiomyocytes. Compared with CT-1 pretreatment groups, the release rate of LDH and cleaved caspase3 protein expression of siRNA interference group were significantly increased. Conclusion The protection role of CT-1 against H/R injury of cardiomyocytes is related to upregulation of HO-1.

关 键 词：心肌细胞 缺氧复氧损伤 心肌营养素-1 血红素加氧酶-1 RNA干扰 

分 类 号：R364.4[医药卫生—病理学]