CAJ | 학술논문

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.
목적 평개결혈후처리대대서심기결혈재관주시선립체심린지합성적영향.방법 웅성SD대서,16～20주령,체중250～ 350 g,건립Langendorff리체심장관주모형.취리체심장64개,채용수궤수자표법,장기분위4조(n=16):정상대조조(C조)、결혈재관주조(Ⅰ/R조)、결혈후처리조(IPO조)화5-간규산+결혈후처리조(5-HD+IPO조).평형관주20 min시,C조계속관주K-H액70 min;Ⅰ/R조관주4℃ST.Thomas정도액10 ml/kg,상온하결혈40 min,재관주37℃함양적K-H액30min;IPO조우결혈40 min시선회복관주10s,결혈10s,반복6차,계지관주37℃함양적K-H액28min;5-HD+IPO조우결혈40 min시관주함100 μmol/L 5-간규산(선립체ATP민감성갑통도조단제)적K-H액5 min,여동IPO조.분별우평형관주20 min(T1)화재관주30 min(T2)시,기록HR、좌실발전압(LVDP)、좌실서장말압(LVEDP)화관맥류량(CF),수집관맥류출액2 ml,측정유산탈경매(LDH)화기산격매(CK)적활성,연후제취선립체,측정심린지함량.결과 여C조비교,기여3조T2시HR、LVDP화CF강저,LVEDP승고,관맥류출액LDH화CK적활성승고,선립체심린지합성강저(P＜0.05);여Ⅰ/R조비교,IPO조T2시HR、LVDP화CF승고,LVEDP강저,관맥류출액LDH화CK적활성강저,선립체심린지함량승고(P＜0.05);여IPO조비교,5-HD+IPO조T2시HR、LVDP화CF강저,LVEDP승고,관맥류출액LDH화CK적활성승고,선립체심린지함량강저(P＜0.05).결론 결혈후처리감경대서심기결혈재관주손상적궤제여기개방선립체ATP민감성갑통도,증가선립체심린지합성유관.
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 for determination of cardiolipin content.Results HR,LVDP,and CF were significantly lower,LVEDP was higher,and the LDH and CK activities in coronary effluent were higher at T2 than at T1 in the four groups.Compared with group C,HR,LVDP and CF were significantly decreased,LVEDP was increased,and the LDH and CK activities in coronary effluent were increased at T2 in the other three groups.Compared with Ⅰ/R group,HR,LVDP and CF were significantly increased,LVEDP was decreased,and the LDH and CK activities in coronary effluent were decreased at T2 in IPO group.Compared with IPO group,HR,LVDP and CF were significantly decreased,LVEDP was increased,and the LDH and CK activities in coronary effluent were increased at T2 in 5-HD+IPO group.Conclusion The mechanism by which ischemic postconditioning reduces myocardial Ⅰ/R injury is related to opening of mitochondrial ATP sensitive potassium channels and increasing mitochondrial cardiolipin synthesis in rats.