参附益心颗粒对心肌梗死后心力衰竭大鼠线粒体动力学的影响

Effect of Shenfu Yixin Granules on Mitochondrial Dynamics in Rats with Heart Failure After Myocardial Infarction

目的:采用左冠状动脉前降支结扎术制备心肌梗死后心力衰竭大鼠模型,观察参附益心颗粒对心力衰竭大鼠线粒体动力学的影响。方法:50只SD雄性大鼠随机取10只为假手术组,其余为造模组,采用左冠状动脉前降支结扎术制备心肌梗死后心力衰竭大鼠模型。根据术后28 d左室射血分数(LVEF)将模型大鼠随机分为模型组、参附益心颗粒低、高剂量组(3.011、15.055 g·kg^(-1))、沙库巴曲缬沙坦钠组(20.83 mg·kg^(-1))。各给药组每日灌胃相应剂量药液,假手术组、模型组灌胃等体积生理盐水,1次/d,连续给药28 d,每组6只。超声检测心功能参数,称量大鼠心脏质量、体质量,计算心脏质量指数,酶联免疫吸附测定法(ELISA)检测血清脑钠肽(BNP)和可溶性生长刺激表达基因2蛋白(sST2)含量,苏木素-伊红(HE)染色观察心肌病理形态,实时荧光定量聚合酶链式反应(Real-time PCR)、蛋白免疫印迹法(Western blot)检测线粒体融合蛋白1/2(Mfn1/2)、视神经萎缩蛋白1(Opa1)、动力相关蛋白1(Drp1)、分裂蛋白1(Fis1)mRNA及蛋白表达。结果:与假手术组比较,模型组的LVEF、Mfn1、Mfn2、Opal mRNA及蛋白水平降低,BNP、sST2、心脏质量指数、Drp1、Fis1 mRNA及蛋白水平增加(P<0.05);与模型组比较,参附益心颗粒高剂量组、沙库巴曲缬沙坦钠组LVEF、Mfn1、Mfn2、Opal mRNA及蛋白表达增高(P<0.05),BNP、sST2、心脏质量指数、Drp1、Fis1 mRNA及蛋白水平降低(P<0.05)。病理观察显示,与假手术组比较,模型组心肌细胞排列紊乱、炎性细胞浸润,心肌纤维化;与模型组比较,各给药组炎性细胞浸润、心肌或肌间质纤维化程度均有改善缓解。结论:参附益心颗粒能抗心力衰竭,降低心脏质量指数、减少BNP、sST2含量,改善心功能,其机制可能与调整线线粒体动力学相关。

Objective:To prepare a rat model of heart failure after myocardial infarction by ligation of the anterior descending branch of the left coronary artery,and to observe the effect of Shenfu Yixin granules on the mitochondrial dynamics of rats with heart failure.Method:Fifty SD male rats were randomly taken ten as the sham operation group and the rest as modeling group.The rat model of heart failure after myocardial infarction was prepared by ligation of anterior descending branch of left coronary artery.According to the left ventricular ejection fraction(LVEF)on the 28^(th)day after operation,the model rats were randomly divided into the model group,Shenfu Yixin granule low-dose and high-dose groups(3.011,15.055 g·kg^(-1))and sacubitril valsartan sodium group(20.83 mg·kg^(-1)).Each administration group was gavaged daily with the corresponding dose of drug solution,while the sham operation group and model group were given the same amount of normal saline once a day for 28 days,with 6 rats in each group.Ultrasound was used to detect the cardiac function parameters,rat heart mass and body mass were weighed to calculate the cardiac mass index,enzyme linked immunosorbent assay(ELISA)was used to detect serum brain natriuretic peptide(BNP)and soluble growth stimulation expressed gene 2 protein(sST2)levels.Hematoxylin-eosin(HE)staining was used to observe the pathological morphology of the myocardium.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and Western blot were used to detect the mRNA and protein expression of mitochondrial fusion protein 1/2(Mfn1/2),optic atrophy protein 1(Opa1),dynamin-related protein 1(Drp1)and fission protein 1(Fis1).Result:Compared with the sham operation group,the mRNA and protein expression of LVEF,Mfn1,Mfn2,Opal in the model group decreased(P<0.05),while BNP,sST2,cardiac mass index,Drp1,Fis1 mRNA and protein levels increased(P<0.05).Compared with the model group,the expression of LVEF,Mfn1,Mfn2,Opal mRNA and protein increased in Shenfu Yixin granule high-dose and sacubitril valsartan sodium groups(P<0.05),while BNP,sST2,cardiac mass index,Drp1,Fis1 mRNA and protein levels decreased(P<0.05).Pathological observation showed that compared with the sham operation group,the model group had disordered arrangement of myocardial cells,inflammatory cell infiltration and myocardial fibrosis.Compared with the model group,the degree of inflammatory cell infiltration,myocardial or interstitial fibrosis was improved and alleviated in all administered groups.Conclusion:Shenfu Yixin granules can resist heart failure,reduce cardiac mass index,decrease BNP and sST2 contents,and improve cardiac function.Its mechanism may be related to the adjustment of mitochondrial dynamics.