蛋白激酶C_ε在缺氧复氧心肌细胞中的调控作用  

Regulatory effect of PKC_ε on hypoxia/reoxygenation injury in cardiomyocytes

作　　者：王锦芝[1] 蔡聪艺[2] 张艳美[3] 石刚刚[3] 

机构地区：[1]汕头大学医学院化学教研室 [2]汕头大学医学院第二附属医院药剂科 [3]汕头大学医学院药理学教研室,广东汕头515041

出　　处：《癌变．畸变．突变》2012年第3期217-221,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金-广东联合基金资助项目(U0932005);广东省自然科学基金项目(10151503102000048);广东省自然科学基金团队项目-潮汕地区特效中药的研制与开发(9351503102000001)

摘　　要：目的:研究蛋白激酶Cε(protein kinase Cε,PKCε)在缺氧复氧(hypoxia/reoxygenation,H/R)所致心肌细胞损伤中的调控作用。方法:取原代培养心肌细胞制作H/R模型,采用Western blot法检测PKCε蛋白转位;双抗体夹心化学发光法和酶联免疫吸附(ELISA)法分别检测培养心肌细胞上清液中肌钙蛋白(cTnI)浓度和肿瘤坏死因子-α(TNF-α)分泌以观察心肌细胞损伤的状况。结果:与正常对照组比较,缺氧2 h复氧30 min时,PKCε总蛋白表达无明显变化,但PKCε由可溶性成分转位至颗粒性成分(P<0.05),延长缺氧时间至4 h,PKCε总蛋白表达明显下降(P<0.05);与H/R组比较,PKCε亚型特异性抑制剂εV1-2可增加cTnI漏出(P<0.05),但对TNF-α的分泌无明显影响。结论:PKCε转位可减轻心肌细胞H/R所致的损伤。OBJECTIVE：We aimed to observe the regulatory effect of protein kinase Cε（PKCε） on injuries in primary cultured cardiomyocytes induced by hypoxia/reoxygenation（H/R）.METHODS：H/R model was made by primary culture of neonatal rat cardiomyocytes.The translocation pattern of PKC activity was assessed by fractionated Western-blot analysis.Measurements including cardiac troponin I（cTnI） release were measured by two-site sandwich chemiluminescence enzyme immunoassay and levels of tumor necrosis factor-α（TNF-α）were measured by enzyme-linked sandwich immunosorbent assay（ELISA） method to assess the degree of injury in cultured cardiomyocytes. RESULTS：Compared with control group,in primary cultured cardiomyocytes exposed to H/R,PKCεtranslocation significantly increased after 2 h of hypoxia and 30 min of reoxygenation（P 〈 0.05）,the protein levels of PKCεdecreased after 4 h of hypoxia（P 〈 0.05）.Compared with H/R group,the PKCεinhibitor peptideεV1-2 increased cTnI release （P 〈 0.05） but did not influence TNF-αsecretion from cardiomyocytes.CONCLUSION：Activation of PKCεcould alleviate the cardiomyocyte cell damage induced by H/R.

关 键 词：心肌细胞 缺氧复氧 蛋白激酶CΕ 

分 类 号：R965[医药卫生—药理学]