健心颗粒对慢性心力衰竭气虚血瘀证模型大鼠心肌能量代谢及PPARα/PGC-1α通路的影响

Effect of Jianxin Granules(健心颗粒)on Myocardial Energy Metabolism and PPARα/PGC-1αPathway in Model Rats of Chronic Heart Failure with Qi Deficiency and Blood Stasis Syndrome

目的从心肌能量代谢角度探讨健心颗粒治疗慢性心力衰竭气虚血瘀证的可能作用机制。方法60只SD大鼠随机分为造模组50只和空白组10只。造模组大鼠采用冠脉结扎配合力竭式游泳、饥饿法建立慢性心力衰竭气虚血瘀证模型。造模成功46只大鼠随机分为模型组10只,缬沙坦组及健心颗粒低、中、高剂量组各9只。缬沙坦组给予7.2 mg/(kg·d)缬沙坦灌胃,健心颗粒低、中、高剂量组分别给予1.6、3.2、6.4g/(kg·d)剂量的健心颗粒灌胃,模型组和空白组每天仅给予1 ml/100 g无菌生理盐水灌胃。各组均每天灌胃2次,共4周。HE和Masson染色观察大鼠心肌组织病理改变。检测大鼠心肌组织能量代谢指标[包括糖原质量分数、脂肪酸(FA)及乳酸浓度、三磷酸腺苷(ATP)浓度]、左室射血分数(EF)、心肌组织过氧化物酶体增殖物激活受体α(PPARα)、过氧化物酶体增殖物激活受体γ辅助活化因子-1α(PGC-1α)、300 KDA腺病毒E1A结合蛋白(P300)、中链酰基辅酶A脱氢酶(MCAD)、腺苷酸转运体(ANT)蛋白及mRNA表达。结果HE和Masson染色结果显示,模型组大鼠心肌细胞重度肥大、排列松散,心肌纤维断裂,成纤维细胞严重增生并伴有炎细胞浸润,心肌纤维化严重,胶原组织增生明显,缬沙坦组和健心颗粒各剂量组心肌组织损伤和胶原组织增生均有不同程度改善。与空白组比较,其余各组大鼠心肌组织FA和乳酸浓度升高,EF、糖原质量分数、ATP浓度和PPARα、PGC-1α、P300、MCAD、ANT mRNA及蛋白表达降低(P<0.05);与模型组比较,缬沙坦组和健心颗粒各剂量组心肌组织FA和乳酸浓度降低,EF、糖原质量分数、ATP浓度和PPARα、PGC-1α、P300、MCAD、ANT mRNA及蛋白表达升高(P<0.05);且健心颗粒高剂量组各指标改善优于缬沙坦组和健心颗粒低、中剂量组(P<0.05),健心颗粒中剂量组各指标改善与缬沙坦组比较差异无统计学意义(P>0.05)。结论健心颗粒可显著改善慢性心力衰竭气虚血瘀证大鼠心肌能量代谢,且以高剂量最佳,推测其可能是通过调控PPARα/PGC-1α通路而实现。

Objective To investigate the potential mechanism of Jianxin Granules(健心颗粒)in the treatment of chronic heart failure(CHF)with qi deficiency and blood stasis syndrome from the perspective of myocardial energy metabolism.Methods Sixty SD rats were randomly divided into modeling group(n=50)and blank group(n=10).The former established the rat model of CHF with qi deficiency and blood stasis syndrome by coronary artery ligation combined with exhaustive swimming and starvation.Forty-six rats were successfully modeled and were divided into model group(n=10),valsartan group(n=9)and Jianxin Granules low-,medium-and high-dose groups(n=9 each)according to the random number table.Valsartan group was given 7.2 mg/(kg·d)of valsartan by gavage,while the low-,medium-and high-dose groups of Jianxin Granules were given 1.6,3.2 and 6.4g/(kg·d)of Jianxin Granules by gavage,respectively.The model group and the blank group were given 1 ml/100 g of sterile normal saline by ga‐vage.The above interventions were administered twice a day for 4 weeks.Hematoxylin eosin(HE)and Masson stain‐ing were used to observe the pathological changes of rat myocardial tissue.The indexes of energy metabolism in rat myocardial tissues including glycogen mass fraction,concentration of fatty acid(FA)and lactic acid,and adenosine triphosphate(ATP)concentration were measured.Left ventricular ejection fraction(EF),peroxisome proliferator ac‐tivated receptorα(PPARα),peroxisome proliferator activated receptorγcoactivator-1α(PGC-1α),300kda adenovi‐rus E1A binding protein(P300),medium chain acyl CoA dehydrogenase(MCAD)and adenylate transporter(ANT)proteins and mRNA expressions were detected.Results HE and Masson staining showed that in the model group,cardiomyocytes were severely hypertrophic and loosely arranged,with broken myocardial fibers,seriously proliferated fibroblasts accompanied by inflammatory cell infiltration,as well as serious myocardial fibrosis and obvious collagen tissue hyperplasia.The myocardial tissues injury and collagen tissue hyperplasia in valsartan group and all three Ji⁃anxin Granules groups were improved in varying degrees.Compared to those in the blank group,the concentration of FA and lactic acid in myocardial tissues increased,and EF,glycogen mass fraction,ATP concentration,and the mRNA and protein expressions of PPARα,PGC-1α,P300,MCAD and ANT decreased in the model group,valsartan group and three Jianxin Granules groups(P<0.05).Compared to those in the model group,the concentration of FA and lactic acid in myocardial tissues decreased,and EF,mass fraction of glycogen,ATP concentration and the mRNA and protein expressions of PPARα,PGC-1α,P300,MCAD and ANT in the valsartan group and Jianxin Gran⁃ules groups increased(P<0.05).More changes were found in the high-dose Jianxin Granules group than the low-and medium-dose groups(P<0.05).The differences were not statistically significant in the mentioned indexes between the medium-doseJianxin Granules group and the valsartan group(P>0.05).Conclusion Jianxin Granules can improve myocardial energy metabolism in CHF rats with qi deficiency and blood stasis syndrome,presumably by regulating PPARα/PGC-1αpathway.And the high-dose Jianxin Granules shows best effect.