缺血后处理对大鼠心肌缺血再灌注时线粒体心磷脂合成的影响：离体实验  

Effect of ischemic postconditioning on mitochondrial cardiolipin synthesis during myocardial ischemia-reperfusion in rats in vitro

作　　者：段忠心[1] 刘兴奎[1] 喻田[1] 

机构地区：[1]遵义医学院麻醉学系,563003

出　　处：《中华麻醉学杂志》2015年第6期680-683,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（30740044）;贵州省科技基金[（2007）2121号]

摘　　要：目的 评价缺血后处理对大鼠心肌缺血再灌注时线粒体心磷脂合成的影响.方法 雄性SD大鼠,16～20周龄,体重250～ 350 g,建立Langendorff离体心脏灌注模型.取离体心脏64个,采用随机数字表法,将其分为4组（n=16）：正常对照组（C组）、缺血再灌注组（Ⅰ/R组）、缺血后处理组（IPO组）和5-羟葵酸＋缺血后处理组（5-HD＋IPO组）.平衡灌注20 min时,C组继续灌注K-H液70 min;Ⅰ/R组灌注4℃ST.Thomas停跳液10 ml/kg,常温下缺血40 min,再灌注37℃含氧的K-H液30min;IPO组于缺血40 min时先恢复灌注10s,缺血10s,反复6次,继之灌注37℃含氧的K-H液28min;5-HD＋IPO组于缺血40 min时灌注含100 μmol/L 5-羟葵酸（线粒体ATP敏感性钾通道阻断剂）的K-H液5 min,余同IPO组.分别于平衡灌注20 min（T1）和再灌注30 min（T2）时,记录HR、左室发展压（LVDP）、左室舒张末压（LVEDP）和冠脉流量（CF）,收集冠脉流出液2 ml,测定乳酸脱氢酶（LDH）和肌酸激酶（CK）的活性,然后提取线粒体,测定心磷脂含量.结果 与C组比较,其余3组T2时HR、LVDP和CF降低,LVEDP升高,冠脉流出液LDH和CK的活性升高,线粒体心磷脂合成降低（P＜0.05）;与Ⅰ/R组比较,IPO组T2时HR、LVDP和CF升高,LVEDP降低,冠脉流出液LDH和CK的活性降低,线粒体心磷脂含量升高（P＜0.05）;与IPO组比较,5-HD＋IPO组T2时HR、LVDP和CF降低,LVEDP升高,冠脉流出液LDH和CK的活性升高,线粒体心磷脂含量降低（P＜0.05）.结论 缺血后处理减轻大鼠心肌缺血再灌注损伤的机制与其开放线粒体ATP敏感性钾通道,增加线粒体心磷脂合成有关.Objective To evaluate the effects of ischemic postconditioning on mitochondrial cardiolipin synthesis during myocardial ischemia-reperfusion （Ⅰ/R） in rats in vitro.Methods Healthy male Sprague-Dawley rats,aged 16-20 weeks,weighing 250-350 g,were used in the study.The animals were anesthetized with intraperitoneal pentobarbital sodium 40 mg/kg and received intraperitoneal heparin 250 U/kg.Their hearts were excised and retrogradely perfused in a Langendorff apparatus.Sixty-four isolated rat hearts were randomly divided into 4 groups （n =16 each） using a random number table：control group （group C）,Ⅰ/R group,ischemic postconditioning group （group IPO） and 5-hydroxydecanoate （5-HD） plus ischemic postconditioning group （group 5-HD ＋ IPO）.After 20 min of equilibration,the hearts were continuously perfused with K-H solution for 70 min in group C.In Ⅰ/R group,after 20 min of equilibration,the hearts were continuously perfused with 4 ℃ ST.Thomas cardioplegic solution 10 ml/kg,and exposed to 40 min of ischemia followed by reperfusion with oxygenated K-H solution at 37 ℃ for 30 min.In group IPO,after 20 min of equilibration,the hearts were subjected to 6 cycles of 10 s reperfusion followed by 10 s ischemia starting from 40 min of ischemia,and then were reperfused with oxygenated K-H solution at 37 ℃ for 28 min.In group 5-HD ＋ IPO,after 20 min of equilibration,the hearts were perfused with K-H solution containing 100 μmol/L 5-HD （mitochondrial ATP-sensitive potassium channel blocker） for 5 min starting from 40 min of ischemia,and then the other procedures were similar to those previously described in group IPO.At 20 min of equilibration （T1） and 30 min of reperfusion （T2）,HR,left ventricular developed pressure （LVDP）,left ventricular end-diastolic pressure （LVEDP） and coronary flow （CF） were recorded.The coronary effluent 2 ml was collected for detection of lactic dehydrogenase （LDH） and creatine kinase （CK） activities.The mitochondria were extracted f

关 键 词：心肌再灌注损伤 线粒体 心磷脂质类 缺血后处理 

分 类 号：R614[医药卫生—麻醉学]