CGRP通过NOS/NO通路抑制低氧诱导的大鼠心肌细胞凋亡  

CGRP inhibits the hypoxia-induced apoptosis via NOS/NO pathway in cardiomyocytes of rat

作　　者：段莉肖 雷寒[1] 张羿[1] 万彪[2] 冯清平[3] 黄玮[1] 

机构地区：[1]重庆医科大学附属第一医院心血管内科,重庆400016 [2]重庆医科大学附属第二医院妇产科,重庆400010 [3]加拿大西安大略大学医学院生理和药理学系

出　　处：《基础医学与临床》2016年第1期30-34,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(81170188;30971212);重庆市自然科学基金(CSCT2009BB5069);国家临床重点专科建设项目(财社[2011]170号)

摘　　要：目的研究降钙素基因相关肽(CGRP)是否通过调节一氧化氮(NO)抑制低氧心肌细胞的凋亡。方法选用H9c2细胞建立低氧损伤模型,CGRP和/或一氧化氮合酶(NOS)抑制剂(L-NAME)预处理细胞,检测低氧后细胞存活率,NOS、NO和凋亡相关蛋白表达水平。结果低氧使心肌细胞存活率降低(P<0.05),诱导性一氧化氮合酶(i NOS)表达明显增加(P<0.001),内皮型一氧化氮合酶(e NOS)和磷酸化内皮型一氧化氮合酶(p-e NOS)表达降低(P<0.001),NO释放减少(P<0.05),P53、caspase-3和细胞色素C(cytochrome C)表达升高(P<0.05);CGRP预处理后,心肌细胞存活率升高(P<0.001),i NOS表达减少(P<0.05),e NOS和p-e NOS表达增加(P<0.05),NO的释放进一步降低(P<0.05),凋亡相关蛋白活性下降(P<0.001);L-NAME处理后,i NOS(P<0.05)和P53(P<0.001)表达降低;L-NAME与CGRP共处理,与CGRP单处理比较,细胞存活率下降(P<0.05),NOS表达降低(P<0.05),P53、caspase-3和cyto C表达升高(P<0.05)。结论 NOS/NO可能介导CGRP对低氧心肌细胞抗凋亡的调控作用。Objective To investigate whether Calcitonin Gene-Related Peptide（ CGRP） plays a protective role in cardiomyocytes against hypoxia-induced apoptosis via an NO-mediated pathway. Methods H9c2 cardiac cells were exposed to hypoxia for 2 h to establish a model of myocardial hypoxic-ischemic injury. The cells were pretreated with either CGRP or an NOS inhibitor（ L-NAME） before being exposed to hypoxia for 30 min. Cell viability was analyzed using a cell counter kit 8（ CCK-8）. The expression levels of several apoptosis proteins（ P53,caspase-3,cytochrome C） and nitric oxide synthase（ NOS） were detected via Western blot assay. An NO kit was used to evaluate the production of NO. Results Our studies demonstrated that pretreatment of H9c2 cardiac cells with CGRP for 30 min prior to exposure to hypoxia markedly improved cell viability; the same effect was observed followingpretreatment with theNOS inhibitor,L-NAME. Additionally,CGRP enhanced the expression of both eNOS and peNOS; the application of both L-NAME and CGRP attenuated the hypoxia-induced expression of iN OS and enhanced the hypoxia-mediated decrease in NO. Furthermore,CGRP significantly decreased the hypoxia-induced overexpression of several apoptotic proteins,P53,caspase-3 and cyto C. Interestingly,the expression levels of cell death,NOS and apoptotic factors were slightly attenuated in the presence of L-NAME and CGRP co-working 2hours of acute hypoxia. Conclusions CGRP protects cardiomyocytes from hypoxia-induced apoptosis via the modulation of the production of NO.

关 键 词：低氧 凋亡 降钙素基因相关肽 一氧化氮 H9C2 

分 类 号：R542.22[医药卫生—心血管疾病]