参附益心颗粒对心肌梗死后心力衰竭大鼠心肌线粒体呼吸功能的影响

Effect of Shenfu Yixin Granule on Myocardial Mitochondrial Respiratory Function in Rats with Heart Failure after Myocardial Infarction

目的观察参附益心颗粒对心力衰竭大鼠线粒体呼吸功能的影响。方法采用结扎大鼠左冠状动脉前降支的方法复制心肌梗死后心力衰竭大鼠模型。将造模成功的40只大鼠随机分为模型组、参附益心颗粒(简称参附)常用剂量组[1.76 g/(kg·d)]、大剂量组[8.8 g/(kg·d)]和氯沙坦组[10 mg/(kg·d)],每组10只,同时设假手术组10只。分别给予相应的药物或相同体积的蒸馏水灌胃4周。利用颈动脉导管法测量相关血流动力学指标:心率(HR)、左室收缩压(LVSP)、左室舒张末压(LVEDP)及左室压最大上升和下降速率(±dp/dtmax);采用Seahorse法检测线粒体呼吸功能相关参数:基础呼吸、最大呼吸、态3呼吸(T3)、态4呼吸(T4)和控制率(RCR);Western Blot法检测心肌线粒体呼吸链复合酶Ⅰ~Ⅴ的蛋白表达量。结果与假手术组比较,模型组HR、LVEDP明显升高(P<0.01),LVSP、±dt/dpmax降低(P<0.05,P<0.01);T3、最大呼吸及RCR1、RCR2值均降低(P<0.01),T4明显升高(P<0.01);呼吸链复合酶Ⅲ和Ⅳ表达降低(P<0.01,P<0.05)。与模型组比较,各药物干预组LVEDP降低(P<0.01),±dt/dpmax升高(P<0.05,P<0.01),参附常用量和大剂量组LVSP增高(P<0.05,P<0.01),参附常用量组和氯沙坦组中HR降低(P<0.05);各药物干预组T3、最大呼吸和RCR值均上升(P<0.01),参附常用量组和氯沙坦组T4下降(P<0.05,P<0.01);各药物干预组呼吸链复合酶Ⅲ均升高(P<0.01,P<0.05),参附大剂量组和氯沙坦组中呼吸链复合酶Ⅳ升高(P<0.05)。结论参附益心颗粒能够改善心衰大鼠线粒体呼吸功能、增加心肌收缩力,其机制可能与抑制线粒体呼吸链复合酶Ⅲ和Ⅳ含量的降低有关。

Objective To observe the effect of Shenfu Yixin Granule on mitochondrial respiratory function in rats with heart failure.Methods The rat model of heart failure after myocardial infarction was established by ligating the anterior descending branch of the left coronary artery.Totally 40 rats were randomly divided into model group,Shenfu Yixin Granule common dose group(1.76 g·kg^-1·d^-1),high dose group(8.8 g·kg^-1·d^-1)and losartan group(10 mg·kg^-1·d^-1),10 in each group.Meanwhile,10 rats were recruited in a sham operation group.The corresponding drugs or distilled water of the equal volume were given intragastrically for 4 weeks.The related hemodynamic indexes were measured by carotid catheterization:heart rate(HR),left ventricular systolic blood pressure(LVSP),left ventricular end diastolic pressure(LVEDP)and left ventricular pressure maximum rise and fall rate(±dp/dtmax).The parameters related to mitochondrial respiratory function were measured by Seahorse method:basic respiration,maximum respiration,state 3 respiration(T3),state 4 respiration(T4)and control rate(RCR).The protein expression of respiratory chain complexⅠ-Ⅴin myocardial mitochondria was detected by Western Blot method.Results Compared with the sham operation group,the HR and LVEDP increased in the model group(P<0.01),LVSP and±dt/dpmax decreased significantly(P<0.05,P<0.01).T3,maximal respiration,RCR1 and RCR2 decreased,T4 increased significantly(P<0.01),and the expression of respiratory chain complex enzymeⅢandⅣdecreased(P<0.01,P<0.05).Compared with the model group,LVEDP decreased and±dt/dpmax increased in each treatment group(P<0.05,P<0.01),and LVSP increased in Shenfu normal dose group and high dose group(P<0.05,P<0.01).HR in Shenfu normal dose group and losartan group decreased(P<0.05).The values of T3,maximum respiration and RCR in each treatment group were significantly increased(P<0.01),T4 respiration decreased significantly in Shenfu normal dose group and losartan group(P<0.05,P<0.01).The respiratory chain complexⅢwas significantly increased in all groups(P<0.01,P<0.05).The respiratory chain complexⅣin Shenfu high dose group and losartan group was significantly higher than that in losartan group(P<0.05).Conclusion Shenfu Yixin Granule could improve mitochondrial respiratory function and increase myocardial contractility in rats with heart failure,which might be related to the inhibition of the content of mitochondrial respiratory chain complex enzymeⅢandⅣ.