摘要
目的研究大鼠心肌缺血再灌注时沉默信息调节因子1(SIRT1)对心肌内质网应激相关凋亡的影响及其与ERK1/2信号通路的关系。方法将大鼠随机分为6组:假手术组、缺血再灌注组、白藜芦醇+缺血再灌注组、白藜芦醇+EX527+缺血再灌注组、白藜芦醇+PD98059+缺血再灌注组、PD98059+缺血再灌注组,每组12只。结扎大鼠冠状动脉左前降支建立大鼠心肌I/R损伤模型。TUNEL法检测心肌细胞凋亡;比色法检测LDH、CK-MB活性。实时荧光定量PCR(qRT-PCR)法检测心肌GRP78、caspase-12和CHOP mRNA的表达;Western印迹检测SIRTl、caspase-12、CHOP、磷酸化ERK1/2和总ERK1/2蛋白的表达。结果 Res+I/R组与I/R组相比,心肌凋亡指数降低(P<0.05),血清LDH及CK-MB活性降低,内质网应激相关凋亡的指标GRP78、caspase-12及CHOP的蛋白表达量和mRNA均降低(P<0.05);给予SIRT1抑制剂后与Res+I/R组相比,以上指标升高;Res+I/R组与I/R组相比,SIRT1及磷酸化ERK1/2蛋白的表达量均增加(P<0.05);而Res+EX+I/R组与Res+I/R组相比,SIRT1、磷酸化ERK1/2蛋白的表达量又降低(P<0.05)。结论 SIRT1能够抑制大鼠在体缺血再灌注心肌的内质网应激凋亡相关蛋白表达,发挥保护心肌的作用,其机制可能与ERK1/2通路激活有关。
Objective To investigate the role of silent information regulator 1 on endoplasmic reticulum dependent apoptosis pathway induced by myocardial ischemic reperfusion and the relationship with ERK1/2 pathway.Methods Rats were divided into 6 groups：control group,ischemic/reperufuison group,resverotrol treatment group,resverotrol plus EX527 （1 μg/kg） treatment group,resverotrol （10 mg/kg） plus PD98059 （0.3 mg/kg） treatment group,PD98059 treatment group,each group n =12.The myocardial I/R injury model in rats was established by ligating left anterior descending coronary artery.TUNEL method was used to detect myocardial apoptosis;Colorimetric method was used to measure LDH and CK-MB activity,qRT-PCR was used to detect the myocardial mRNA expression of GRP78,caspase-12 and CHOP;The protein expression of sirt1,caspase-12,CHOP,phosphor-ERK1/2 and total ERK1/2 were measured by westernblot.Results Compared with I/R group,the myocardial cell apoptosis index,LDH and CK-MB activity,mRNA expression of GRP78,caspase-12,CHOP and the protein expression of caspase12,CHOP decreased while the protein expression of SIRT1 and phosphor-ERK1/2 increased in resveratrol treatment group.Then treated with SIRT1 inhibitor EX527 in Res ＋ EX ＋ I/R group the increase or decrease result of those index were conteracted compared with Res ＋ I/R group.Conclusions SIRT1 can protect heart from ischemic reperfusion injury through inhibiting ER-dependent apoptosis protein expression,the mechanism of which is partly related with ERK1/2 pathway activation.
出处
《基础医学与临床》
CSCD
2015年第6期761-766,共6页
Basic and Clinical Medicine
基金
国家临床重点专科[财社(2011)170号]
重庆市医学重点学科[渝卫科教(2007)2号]
重庆市卫生局医学科研计划重点项目(2012-1-022)
