CAJ | 학술논문

目的探讨七氟烷预处理对大鼠离体心肌缺血/再灌注损伤(ischemia/reperfusion injury,I/RI)时心肌细胞凋亡及线粒体细胞色素C释放的影响.方法 40只SD大鼠应用随机数字表法随机分为4组(每组10只):对照组(C组),0.75％、1.5％、3.0％七氟烷预处理组(S1、S2、S3组).应用Langendorff离体心脏灌注模型,经主动脉用Krebs-Henseleit( K-H)液逆行灌注.各组均缺血30 min,再灌注60 min.S1、S2、S3组在缺血前分别以含相应浓度七氟烷的K-H液灌注10 min,再冲洗5 min,重复2次.记录平衡灌注末、缺血前即刻、再灌注30、60 min时的心功能指标.再灌注60 min时用DNA原位末端缺口标记技术(terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling,TUNEL)测定凋亡细胞,提取心肌线粒体,免疫印迹法测定线粒体和胞浆的细胞色素C水平.结果与C组比较,S1、S2、S3组再灌注30、60 min时左室舒张末压(left ventricular end-diastolic pressure,LVEDP)降低、左室发展压(left ventricular developed pressure,LVDP)升高(P＜0.05);与C组比较,S1、S2、S3组凋亡率明显下降(P＜0.05或0.01),再灌注末心肌细胞凋亡率C、S1、S2、S3组分别为(33.10±2.57)％、(28.03±3.11)％、(25.20±3.04)％、(24.06±2.58)％;与C组比较,S1、S2、S3组心肌线粒体细胞色素C释放减少,胞浆细胞色素C的量明显降低(P＜0.05或0.01).结论七氟烷预处理能够通过抑制心肌线粒体细胞色素C释放到胞浆,降低心肌细胞凋亡率,减轻心肌I/RI.
목적 탐토칠불완예처리대대서리체심기결혈/재관주손상(ischemia/reperfusion injury,I/RI)시심기세포조망급선립체세포색소C석방적영향.방법 40지SD대서응용수궤수자표법수궤분위4조(매조10지):대조조(C조),0.75％、1.5％、3.0％칠불완예처리조(S1、S2、S3조).응용Langendorff리체심장관주모형,경주동맥용Krebs-Henseleit( K-H)액역행관주.각조균결혈30 min,재관주60 min.S1、S2、S3조재결혈전분별이함상응농도칠불완적K-H액관주10 min,재충세5 min,중복2차.기록평형관주말、결혈전즉각、재관주30、60 min시적심공능지표.재관주60 min시용DNA원위말단결구표기기술(terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling,TUNEL)측정조망세포,제취심기선립체,면역인적법측정선립체화포장적세포색소C수평.결과 여C조비교,S1、S2、S3조재관주30、60 min시좌실서장말압(left ventricular end-diastolic pressure,LVEDP)강저、좌실발전압(left ventricular developed pressure,LVDP)승고(P＜0.05);여C조비교,S1、S2、S3조조망솔명현하강(P＜0.05혹0.01),재관주말심기세포조망솔C、S1、S2、S3조분별위(33.10±2.57)％、(28.03±3.11)％、(25.20±3.04)％、(24.06±2.58)％;여C조비교,S1、S2、S3조심기선립체세포색소C석방감소,포장세포색소C적량명현강저(P＜0.05혹0.01).결론 칠불완예처리능구통과억제심기선립체세포색소C석방도포장,강저심기세포조망솔,감경심기I/RI.
Objective To explore the effect of sevoflurane preconditioning on cardiocyte apoptosis and cytochrome C release from mitochondria during ischemia/reperfusion in isolated rat hearts.Methods Forty male Sprague-Dawley rats weighing 250 g-300g were randomly divided into 4 groups (n=10 for each group):control group (C); 3 different concentrations of sevoflurane preconditioning groups of 0.75％ (S1 ),1.5％ (S2),3.0％ (S3) sevoflurane respectively.All of the isolated rat hearts were retrogradely perfused via aorta with Krebs-Henseleit (K-H) solution on Langendorff apparatus.The isolated hearts were subjected to ischemia for 30 min followed by 60 min reperfusion.Hearts in the S1,S2,S3 groups were preconditioned by perfusing with K-H solution which containing 0.75％,1.5％,3.0％ sevoflurane respectively for 10 min and then followed by 5 min K-H solution washout before ischemia.The preconditioning procedure was repeated twice.Hemodynamics of the hearts were recorded after equilibration (baseline values),immediately before ischemia,30 and 60 min after reperfusion respectively.Apoptotic myocardial cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) and the level of cytochrome C expression in myocardial cytosol and mitochondria was measured by Western Blot at the end of reperfusion.Results At the end of 30 and 60 min reperfusion,left ventricular end-diastolic pressure (LVEDP) was significantly lower and left ventricular developed pressure (LVDP)was significantly higher in S1,S2,S3 groups than those in C group (P＜0.05).Apoptotic index decreased significantly in S1,S2,S3groups compared with C group at the end of reperfusion (P＜0.05,P＜0.01) and the apoptotic index of C,S1,S2,S3 groups was (33.10±2.57)％,(28.03±3.11)％,(25.20±3.04)％,(24.06±2.58)％ respectively.Cytochrome C level increased significantly in mitochondria but decreased significantly in cytosol in S1,S2,S3 groups as compared with C group (P＜0.05,P＜0.01 ).Conclusions Sevoflurane preconditioning decreased cardiocyte apoptosis,protected the heart against ischemia/reperfusion injury by attenuatiing the release of cytochrome C from mitochondria to cytosol.