中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2010年
7期
850-854
,共5页
杨柳青%喻田%刘兴奎%余志豪
楊柳青%喻田%劉興奎%餘誌豪
양류청%유전%류흥규%여지호
吡那地尔%器官保存液%线粒体,心脏
吡那地爾%器官保存液%線粒體,心髒
필나지이%기관보존액%선립체,심장
Pinacidil%Organ preservation solution%Mitochondria,heart
目的 探讨含吡那地尔的心脏保存液(HTK液)对大鼠离体心脏线粒体呼吸功能的影响.方法 健康清洁级SD大鼠120只,建立离体心脏Langendorff灌注模型,随机分为5组,HTK组(Ⅰ组)、吡那地尔+HTK液组(Ⅱ组)、吡那地尔+HTK液+5-羟葵酸(5-HD)组(Ⅲ组)、吡那地尔+HTK液+HMR-1098组(Ⅳ组)、吡那地尔+HTK液+HMR-1098+5-HD组(Ⅴ组).K-H液平衡灌注10 min后灌注心脏停搏液:Ⅰ组灌注HTK液;Ⅱ组灌注含吡那地尔0.5 mmol/L的HTK液;Ⅲ组灌注含吡那地尔0.5 mmol/L和5-HD 100μmol/L的HTK液;Ⅳ组灌注含吡那地尔0.5 mmol/L和HMR-1098 100μmol/L的HTK液;Ⅴ组灌注含吡那地尔0.5 mmol/L、5-HD 100μmol/L和HMR-1098 100 μmol/L的HTK液.停搏后取心脏保存于4℃各组相应液体中8 h,然后用37 ℃含氧K-H液再灌注60 min.于平衡灌注10 min(T1)、保存8 h(T2)、复灌60 min(T3)时测定线粒体呼吸功能[3态呼吸(S3)和4态呼吸(S4)的耗氧速率、呼吸控制率(RCR)及磷氧比(P/O)].于T1和T3时收集冠脉流出液测定心肌肌钙蛋白I(cTnI)浓度、肌酸激酶同功酶(CK-MB)及乳酸脱氢酶(LDH)的活性.电镜下观察心肌细胞超微结构.结果 与Ⅰ组比较,Ⅱ组～Ⅳ组RCR、P/O、S3耗氧速率升高,cTnI、CK-MB和LDH的水平降低(P＜0.05).与Ⅱ组比较,Ⅲ组～Ⅴ组RCR、P/O、S3耗氧速率降低,cTnI、CK-MB和LDH的水平升高(P＜0.05).与Ⅲ组比较,Ⅳ组RCR、P/O、S3耗氧速率升高,cTnI、CK-MB和LDH的水平降低,Ⅴ组RCR、P/O、S3耗氧速率降低,cTnI、CK-MB和LDH的水平升高(P＜0.05).与Ⅳ组比较,V组RCR、P/O、S3耗氧速率降低,cTnI、CK-MB和LDH水平升高(P＜0.05).各时点S4耗氧速率组间比较差异无统计学意义(P＞0.05).Ⅱ组心肌细胞损伤较轻,Ⅲ组和Ⅳ组损伤程度相近但重于Ⅱ组,Ⅰ组和Ⅴ组损伤最重.结论 含吡那地尔的HTK液可改善心脏线粒体呼吸功能,减轻心肌损伤,提高心脏保存效果,线粒体ATP敏感性钾通道和细胞膜ATP敏感性钾通道均参与了吡那地尔的心肌保护效应,且线粒体ATP敏感性钾通道发挥主导作用.
目的 探討含吡那地爾的心髒保存液(HTK液)對大鼠離體心髒線粒體呼吸功能的影響.方法 健康清潔級SD大鼠120隻,建立離體心髒Langendorff灌註模型,隨機分為5組,HTK組(Ⅰ組)、吡那地爾+HTK液組(Ⅱ組)、吡那地爾+HTK液+5-羥葵痠(5-HD)組(Ⅲ組)、吡那地爾+HTK液+HMR-1098組(Ⅳ組)、吡那地爾+HTK液+HMR-1098+5-HD組(Ⅴ組).K-H液平衡灌註10 min後灌註心髒停搏液:Ⅰ組灌註HTK液;Ⅱ組灌註含吡那地爾0.5 mmol/L的HTK液;Ⅲ組灌註含吡那地爾0.5 mmol/L和5-HD 100μmol/L的HTK液;Ⅳ組灌註含吡那地爾0.5 mmol/L和HMR-1098 100μmol/L的HTK液;Ⅴ組灌註含吡那地爾0.5 mmol/L、5-HD 100μmol/L和HMR-1098 100 μmol/L的HTK液.停搏後取心髒保存于4℃各組相應液體中8 h,然後用37 ℃含氧K-H液再灌註60 min.于平衡灌註10 min(T1)、保存8 h(T2)、複灌60 min(T3)時測定線粒體呼吸功能[3態呼吸(S3)和4態呼吸(S4)的耗氧速率、呼吸控製率(RCR)及燐氧比(P/O)].于T1和T3時收集冠脈流齣液測定心肌肌鈣蛋白I(cTnI)濃度、肌痠激酶同功酶(CK-MB)及乳痠脫氫酶(LDH)的活性.電鏡下觀察心肌細胞超微結構.結果 與Ⅰ組比較,Ⅱ組～Ⅳ組RCR、P/O、S3耗氧速率升高,cTnI、CK-MB和LDH的水平降低(P＜0.05).與Ⅱ組比較,Ⅲ組～Ⅴ組RCR、P/O、S3耗氧速率降低,cTnI、CK-MB和LDH的水平升高(P＜0.05).與Ⅲ組比較,Ⅳ組RCR、P/O、S3耗氧速率升高,cTnI、CK-MB和LDH的水平降低,Ⅴ組RCR、P/O、S3耗氧速率降低,cTnI、CK-MB和LDH的水平升高(P＜0.05).與Ⅳ組比較,V組RCR、P/O、S3耗氧速率降低,cTnI、CK-MB和LDH水平升高(P＜0.05).各時點S4耗氧速率組間比較差異無統計學意義(P＞0.05).Ⅱ組心肌細胞損傷較輕,Ⅲ組和Ⅳ組損傷程度相近但重于Ⅱ組,Ⅰ組和Ⅴ組損傷最重.結論 含吡那地爾的HTK液可改善心髒線粒體呼吸功能,減輕心肌損傷,提高心髒保存效果,線粒體ATP敏感性鉀通道和細胞膜ATP敏感性鉀通道均參與瞭吡那地爾的心肌保護效應,且線粒體ATP敏感性鉀通道髮揮主導作用.
목적 탐토함필나지이적심장보존액(HTK액)대대서리체심장선립체호흡공능적영향.방법 건강청길급SD대서120지,건립리체심장Langendorff관주모형,수궤분위5조,HTK조(Ⅰ조)、필나지이+HTK액조(Ⅱ조)、필나지이+HTK액+5-간규산(5-HD)조(Ⅲ조)、필나지이+HTK액+HMR-1098조(Ⅳ조)、필나지이+HTK액+HMR-1098+5-HD조(Ⅴ조).K-H액평형관주10 min후관주심장정박액:Ⅰ조관주HTK액;Ⅱ조관주함필나지이0.5 mmol/L적HTK액;Ⅲ조관주함필나지이0.5 mmol/L화5-HD 100μmol/L적HTK액;Ⅳ조관주함필나지이0.5 mmol/L화HMR-1098 100μmol/L적HTK액;Ⅴ조관주함필나지이0.5 mmol/L、5-HD 100μmol/L화HMR-1098 100 μmol/L적HTK액.정박후취심장보존우4℃각조상응액체중8 h,연후용37 ℃함양K-H액재관주60 min.우평형관주10 min(T1)、보존8 h(T2)、복관60 min(T3)시측정선립체호흡공능[3태호흡(S3)화4태호흡(S4)적모양속솔、호흡공제솔(RCR)급린양비(P/O)].우T1화T3시수집관맥류출액측정심기기개단백I(cTnI)농도、기산격매동공매(CK-MB)급유산탈경매(LDH)적활성.전경하관찰심기세포초미결구.결과 여Ⅰ조비교,Ⅱ조～Ⅳ조RCR、P/O、S3모양속솔승고,cTnI、CK-MB화LDH적수평강저(P＜0.05).여Ⅱ조비교,Ⅲ조～Ⅴ조RCR、P/O、S3모양속솔강저,cTnI、CK-MB화LDH적수평승고(P＜0.05).여Ⅲ조비교,Ⅳ조RCR、P/O、S3모양속솔승고,cTnI、CK-MB화LDH적수평강저,Ⅴ조RCR、P/O、S3모양속솔강저,cTnI、CK-MB화LDH적수평승고(P＜0.05).여Ⅳ조비교,V조RCR、P/O、S3모양속솔강저,cTnI、CK-MB화LDH수평승고(P＜0.05).각시점S4모양속솔조간비교차이무통계학의의(P＞0.05).Ⅱ조심기세포손상교경,Ⅲ조화Ⅳ조손상정도상근단중우Ⅱ조,Ⅰ조화Ⅴ조손상최중.결론 함필나지이적HTK액가개선심장선립체호흡공능,감경심기손상,제고심장보존효과,선립체ATP민감성갑통도화세포막ATP민감성갑통도균삼여료필나지이적심기보호효응,차선립체ATP민감성갑통도발휘주도작용.
Objective To investigate the effect of heart preservation solution containing pinacidil on mitochondrial function in isolated rat hearts. Methods One hundred and twenty pathogen-free SD rats of both sexes weighing 250-350 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital 65 mg/kg. Their hearts were immediately removed and perfused in a Langendorff apparatus. Left ventricular enddiastolic pressure was measured from a fluid-filled latex balloon inserted in the left ventricle. The isolated hearts were randomized into 5 groups (n = 24 each):group Ⅰ was perfused with cardioplegic solution HTK; group Ⅱ with HTK containing pinacidil (a non-specific sarcKATP and mitoKATP channel opener) 0.5 mmol/L; group Ⅲ with HTK containing pinacidil 0.5 mmol/L + 5-HD (a selective mitoKATP channel blocker) 100 μmol/L; group Ⅳ with HTK containing pinacidil 0.5 mmol/L + HMR-1098 100 μmol/L (a selective sarcKATP channel blocker) and group Ⅴ with HTK containing pinacidil 0.5 mmol/L + 5-HD 100 μmol/L + HMR-1098 100μmol/L. The isolated hearts were perfused with simple HTK or HTK containing pinacidil or pinacidil + 5-HD and/or HMR 20 ml/kg at 10 ml/min and then removed from Langendorff apparatus and dipped into the same HTK solution for 8 h at 4 ℃followed by 60 min reperfusion. The respiratory function of mitochondria (respiratory control rate (RCR), the rate of oxygen consumption in state 3/state 4 and P/O) was measured at the end of equilibration (T1) after 8 hpreservation (T2) and at the end of 60 min reperfusion (T3). The CK-MB and LDH activities and cTnI expression in myocardium was detected at T1 and T3. The ultrastructure of myocardium was examined at T3. Results Perfusion suspension-reperfusion (PS/R) significantly decreased mitochondrial respiratory function (RCR, P/O and rate of O2 consumption in state 3) and increased myocardial cTnI concentration and CK-MB and LDH activities at T3 compared with baseline at T1 in group Ⅰ. Pinacidil significantly increased mitochondrial respiratory function (RCR, P/O and rate of O2 consumption in state 3) and decreased myocardial cTnI concentration and CK-MB and LDH activities in group Ⅱ as compared with group Ⅰ-indicative of protective effect of pinacidil on mitochondria against PS/R injury. The protective effect of pinacidil against PS/R injury was attenuated by 5-HD and/or HMR1098. The myocardial damage was slightest in group Ⅱ . Conclusion Both sarcolemmal and mitochondrial KATPchannel are involved in the protective effect of pinacidil against PS/R-induced myocardial damage during heart preservation.
