PKCε信号通路介导槲皮素拮抗心肌细胞缺氧/复氧损伤  

Protective effect of quercetin on rat cardiomyocytes against anoxia /reoxygenation injury is mediated by PKCε signaling pathway

作　　者：汤蕾[1,2] 彭易安[1,2] 胥甜甜[2] 易小清[2] 刘瑛[2] 罗喻超[2] 尹东[3] 何明[1,2] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330047 [2]南昌大学药学院药理学与分子治疗学教研室,江西南昌330006 [3]南昌大学第二附属医院分子医学中心实验室,江西南昌330006

出　　处：《中国病理生理杂志》2013年第9期1567-1572,共6页Chinese Journal of Pathophysiology

基　　金：国家973计划项目(No.2009CB526405);国家自然科学基金资助项目(No.81260492)

摘　　要：目的:探讨槲皮素拮抗心肌细胞缺氧/复氧(A/R)损伤与蛋白激酶Cε(PKCε)之间的关系。方法:培养新生大鼠原代心肌细胞,构建A/R模型,Western blotting检测槲皮素对PKCε表达水平的调节。检测A/R损伤后各组细胞乳酸脱氢酶活性、肌酸激酶活性、细胞存活率、活性氧(ROS)含量、线粒体膜电位、线粒体通透性转换孔(mPTP)的开放和细胞凋亡。结果:A/R前72 h加入40μmol/L槲皮素可明显上调心肌细胞PKCε蛋白表达水平,并显著提高细胞存活率,减少ROS产生,维持线粒体膜电位,抑制mPTP开放,减少细胞凋亡(P<0.01)。同时经槲皮素和PKCε抑制剂εV1-2预处理后,则槲皮素的上述保护作用均明显减弱或消失(P<0.01)。结论:槲皮素可以上调心肌细胞PKCε蛋白表达水平,其抗心肌缺血再灌注损伤的保护作用机制可能依赖于PKCε途径。AIM：To investigate whether quercetin （Que） protects cardiomyocytes from anoxia/reoxygenation （A/R） injury through protein kinase C epsilon （PKCε） pathway. METHODS：Primary cardiomyocytes were isolated from neonatal SD rats and exposed to A/R （3 h of anoxia followed by 2 h of reoxygenation） as well as Que and/or εV1-2 （a selective PKCε inhibitor） preconditioning. The expression of PKCε in the cells was detected by Western blotting. The levels of lactate dehydrogenase （LDH） and creatine kinase （CK） in cell culture supernatants, the reactive oxygen species （ROS） and mitochondrial membrane potential in the cells, the opening of mitochondrial permeability transition pore （mPTP） and the cell viability and apoptosis were also measured. RESULTS：The expression of PKCε protein was significantly increased in the cardiomyocytes pretreated with 40 μmol/L Que 72 h before A/R （P〈0.01 vs A/R group）. Meanwhile, Que preconditioning could increase cell survival rate, decrease ROS production and cell apoptosis, alleviate the loss of mitochondrial membrane potential and inhibit the opening of mPTP induced by A/R injury （P〈0.01 vs A/R group）. However, pretreatment with Que and εV1-2 attenuated these protective effects of Que （P〈0.01 vs Que＋A/R group）. CONCLUSION：One of the mechanisms underlying the cardioprotective effect of Que might be the increase in PKCε protein expression and the activation of its downstream pathway.

关 键 词：槲皮素 蛋白激酶CΕ 心肌保护 缺氧 复氧 

分 类 号：R965.1[医药卫生—药理学]