缺氧预处理对离体心肌细胞膜和线粒体缝隙连接蛋白43表达的影响  

The effects of hypoxic preconditioning on connexin 43 expression of cell membrane and mitochondria in isolated myocardial cells

作　　者：刘人宁[1] 曾志羽[1] 钟国强[1] 何燕[1] 柯红红[1] 周柳荣[1] 

机构地区：[1]广西医科大学第一附属医院心血管内科,南宁530021

出　　处：《广东医学》2013年第4期509-511,共3页Guangdong Medical Journal

基　　金：广西自然科学基金资助项目(编号:2010GXNSFA013167)

摘　　要：目的通过建立离体心肌细胞缺氧/复氧模型,探讨缺血预处理(IPC)对心肌细胞膜和线粒体缝隙连接蛋白43(Cx43)表达的影响,阐明IPC抗缺血再灌注损伤的机制。方法将培养72 h的SD大鼠乳鼠原代细胞随机分为空白对照组、缺氧/复氧再灌注组(HR组)及缺氧预处理组(HPC组),使用实时荧光定量PCR检测Cx43 mRNA水平,Western blot半定量细胞膜、线粒体的Cx43及磷酸化Cx43水平。结果 HPC组的Cx43 mRNA、细胞膜和线粒体的Cx43及磷酸化Cx43均明显高于HR组(P<0.01),与空白对照组比较差异无统计学意义(P>0.05)。结论 IPC可以保持心肌细胞Cx43 mRNA和细胞膜、线粒体蛋白Cx43的表达水平,减少由缺血再灌注引起的Cx43去磷酸化,增加磷酸化Cx43的含量,从而有效地减少缺血再灌注对心肌的损伤。Objective To investigate the effects of hypoxic preconditioning on connexin 43 （ Cx43 ） expression of cell membrane and mitochondria in isolated myocardial cells by establishing the model of hypoxic/reoxygenation; thus toclarify the mechanism of ischemic preconditioning （IPC） on ischemia - reperfusion （I/R） injury. Methods Primary myocardial cells of Sprague - Dawley （SD） rats cultured for 72h were randomly divided into 3 groups： control group, hy-poxia reperfusion （HR） group, and hypoxia preconditioning （HPC） group. The mRNA levels of Cx43 were analyzed using FQ - PCR. The expression of Cx43 and phosphorylated Cx43 in both cell membrane and mitochondria, were meas-ured by Western blot analysis. Results The levels of Cx43 mRNA, Cx43 and phosphorylated Cx43 in both cell mem- brane and mitochondria, were significantly higher in HPC group than those in HR group （P 〈 0. 01 ）. There was no signif-icant difference between HPC group and control group （P 〉 0. 05）. Conclusion Ischemic preconditioning attenuates the I/R -induced down -regulation of Cx43mRNA, Cx43 and phosphorylated Cx43 in both cell membrane and mitochondria,thus protects the I/R cardiac injury.

关 键 词：缺氧预处理 缺血预处理 缺血再灌注 线粒体 细胞膜 缝隙连接蛋白43 

分 类 号：R978.15[医药卫生—药品]