法舒地尔对大鼠离体心脏缺血/再灌注损伤的保护作用

Cardioprotective effects of fasudil in myocardial ischemia-reperfusion injury in rats

目的观察盐酸法舒地尔对大鼠离体心脏缺血/再灌注(I/R)损伤是否有保护作用。方法SD大鼠19只,随机分为3组:I/R组、I/R+F组和对照组。用改良的Langendorff灌流装置,用K-H液行主动脉逆行灌流,建立大鼠离体心脏I/R损伤实验模型。I/R组预灌流20 min,停灌45 min,再灌30 min;I/R+F组于再灌注时在灌流液中加入盐酸法舒地尔注射液(10 mg/kg);对照组连续灌流95 min。连续记录左心室收缩功能曲线,收集冠脉流出液,检测冠脉流出液中乳酸脱氢酶(LDH)、肌酸激酶(CK)、肌红蛋白(Mb)漏出量以及心肌细胞内钙、心肌组织一氧化氮(NO)含量和髓过氧化物酶(MPO)活力。结果心肌缺血使冠脉流出量减少,LDH、CK、Mb增加,再灌注后冠脉流出量进一步减少,LDH、CK、Mb进一步增加,同时增加细胞内钙,增加MPO活力,减少NO生成。盐酸法舒地尔逆转再灌注后冠脉流出量减少和LDH、CK和Mb漏出增加,降低细胞内钙、MPO活力,逆转NO生成减少。I/R使左室发展峰压平均值、平均±dp/dtmax均下降,盐酸法舒地尔对左室发展峰压的改变无明显影响,但改善±dp/dtmax降低。结论盐酸法舒地尔对心肌I/R损伤有保护作用,增强I/R引起的"无复流"现象和心肌收缩能力降低的恢复,此作用与逆转NO生成减少、MPO活性增高和细胞内钙超载等因素有关。

AIM To determine whether fasudil （a specific Rho-kinase inhibitor） exerts cardioprotective effect on myocardial ischemia/reperfusion（I/R） injury in isolated heart of rats. METHODS Nineteen SD rats were randomly divided into three groups- I/R group （ n = 7 ）, I/R ＋ F group （n = 6） and control group （n = 6）. The model of isolated myocardial I/R injury was prepared using the improved Langendorff apparatus, by preperfusion for 20 min, followed by a suspension for 45 min and reperfusion for 30 min with Krebs-Henselert （K-H） solution. Fasudil hydrochloride injection （10 mg/Kg） was added to K-H solution during reperfusion in I/R ＋ F group and the perfusion was kept until 95 min in control group. The isovolumic left ventricular pressure was recorded in time with a water-filled latex balloon attached to a transducer which was connected to a biosignal collecting and analyzing system. Coronary effluent was collected and the leakage of lactate dehydrogenase （ LDH）, and creatine kinase （CK） and myoglobin （Mb） were measured in coronary effluent. Myocardial nitric oxide concentration, myeloperoxidase activity and intracellular calcium were also detected. RESULTS Fasudil reversed I/R induced decrease of coronary effluent and increase of the leakage of LDH, CK and Mb, led to a 4-fold increase in nitric oxide and reduced myeloperoxidase activity and intracellular calcium. Fasudil also significantly improved the left ventricular contraction function. In I/R ＋ F group, the recovery of dp/dtmax during reperfusion was remarkably better than that in I/R group. CONCLUSION These results indicate that inhibition of Rho kinase activity during reperfusion exerts cardioprotective effects and enhances the recovery of ＂no reflow＂ phenomenon and myocardial contractile function, possibly by nitric oxide generation and bioavailability.