摘要
目的探讨氢盐水药物后适应对急性心肌梗死大鼠心肌细胞呼吸链复合物Ⅲ活性和线粒体膜电位的影响。方法 64只SD大鼠随机分为假手术组、缺血再灌注(reperfusion/injury,R/I)组、氢盐水(saturated hydrogen saline,Hyd)组和抗霉素A(antimycin A,Ant)组共4组。急性心肌缺血45 min后,于再灌注即刻根据分组给予相应药物干预。再灌注5 min后处死大鼠,采用硫代巴比妥酸法和分光光度计法测定缺血心肌组织氧自由基、采用分光光度法线粒体复合物Ⅲ活性、采用分光光度法测定线粒体膜电位,采用实时荧光定量聚合酶链反应(quantitative real time polymerase chain reaction,qPCR)法测定线粒体中细胞色素(cytochrome,Cyt)-b的mRNA浓度。另24 h后处死大鼠,使用Western Blot法测定线粒体中Cyt-b浓度,胞浆中Cyt-c浓度,使用TUNNEL法测定心肌细胞凋亡指数。结果再灌注5 min后,Hyd组微量丙二醛(malondialdehyde,MDA)和羟自由基浓度显著低于R/I组(P<0.05),超氧化物歧化酶(superoxide dismutase,SOD)浓度高于R/I组,差异有统计学意义(P<0.05)。Hyd组复合物Ⅲ活性[(3.52±0.31)U/mgpro vs.(1.43±0.69)U/mgpro,P<0.05]、线粒体膜电位(0.99±0.09 vs. 0.65±0.11,P<0.05),Cyt-b表达量(再灌注5 min后线粒体qPCR:0.82±0.09 vs. 0.41±0.13,P<0.05;再灌注24 h后线粒体Western Blot:0.60±0.09 vs. 0.37±0.08,P<0.05)均显著高于R/I组,胞浆中Cyt-c表达量、凋亡指数显著低于R/I组,差异有统计学意义(P<0.05)。Ant组Cyt-b表达量、复合物Ⅲ活性与Hyd组比较,差异无统计学意义(P>0.05)。Ant组氧自由基浓度、线粒体膜电位显著低于Hyd组;Cyt-c表达量和凋亡指数显著高于Hyd组,且以上指标均显著优于R/I组,差异有统计学意义(P<0.05)。结论氢盐水药物后适应可显著改善大鼠急性心肌梗死再灌注后心肌细胞复合物Ⅲ活性和线粒体膜电位,继而减轻心肌细胞凋亡;Ant能部分抵消这种保护作用。
Objectives To study the influence of hydrogen saline pharmacological postconditioning on complex Ⅲactivity and mitochondrial transmembrane potential in acute myocardial infarction rats. Methods Totally 64 SD rats were randomly divided into 4 groups:Sham,reperfusion/injury(R/I),saturated hydrogen saline(Hyd)and antimycin A(Ant)group. After 45 min of myocardial ischemia,at the time of reperfusion different drugs were administrated according to different groups using micro syringe. After 5 min reperfusion,the rats were sacrificed. Oxygen radicals in ischemic myocardial tissue were measured by thiobarbituric acid method and spectrophotometer method. Mitochondria transmembrane potential was tested by spectrophotometer method. Concentration of cytochrome(Cyt)-b mRNA was measured by quantitative real time polymerase chain reaction(qPCR). After 24 h reperfusion,another rats were sacrificed. Concentrations of Cyt-b in mitochondria and Cyt-c in endochylema were tested by Western Blot. Apoptotic index was tested by TUNNEL method. Results After 5 min of reperfusion,the concentrations of malondialdehyde(MDA)and hydroxyl radical(OH-)were significantly lower,concentration of SOD was significantly higher than those in R/I group(P<0.05). At the same time,complex Ⅲ activity[(3.52±0.31)U/mgpro vs.(1.43±0.69)U/mgpro,P<0.05],mitochondrial transmembrane potential(0.99 ± 0.09 vs. 0.65 ± 0.11,P<0.05),expression of Cyt-b(qPCR:0.82±0.09 vs. 0.41±0.13,P<0.05;Western Blot:0.60±0.09 vs. 0.37±0.08,P<0.05)in Hyd group were all significantly higher than those in R/I group. The expression of Cyt-c and apoptotic index in Hyd group were significantly lower than those in R/I group(P<0.05). Apart from expression of Cyt-b and complex Ⅲ activity(P>0.05),other varibles of Ant group were all significantly worse than those of Hyd group and much better than those of R/I group(P<0.05).Conclusions Hydrogen asline pharmacological postconditioning could improve complex Ⅲ activity and mitochondrial transmembrane potential,alleviate myocardial apoptosis in rats,but Ant could partly counteract the protection.
作者
张国明
孙媛媛
王斌
叶涛
戴翠莲
丁立成
王焱
ZHANG Guo-ming;SUN Yuan-yuan;WANG Bin;YE Tao;DAI Cui-lian;DING Li-cheng;WANG Yan(Department of Cardiology,Xiamen Cardiovascular Hospital,Xiamen,Fujian 361009,China)
出处
《岭南心血管病杂志》
2019年第6期697-703,共7页
South China Journal of Cardiovascular Diseases
基金
福建省自然科学基金（项目编号：2018D0020）
厦门市科技局惠民项目（项目编号：3502Z20174006）
关键词
心肌梗死
再灌注损伤
线粒体膜电位
复合物Ⅲ
药物后适应
氢盐水
大鼠
myocardial infarction
reperfusion injury
mitochondria transmembrane potential
complex Ⅲ
pharmacological postconditioning
hydrogen saline
rat
