温阳益气活血方对慢性心力衰竭大鼠心肌组织Na^(+)-K^(+)-ATP酶、Ca^(2+)-ATP酶的影响

Effects of Wenyang Yiqi Huoxue Prescription on Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase in Myocardial Tissue of Rats with Chronic Heart Failure

目的观察温阳益气活血方对慢性心力衰竭大鼠心肌组织Na^(+)-K^(+)-ATP酶、Ca^(2+)-ATP酶的影响,探讨其治疗心力衰竭的作用机制。方法采用皮下多点注射异丙肾上腺素制备慢性心力衰竭大鼠模型,将成模大鼠随机分为模型组、西药组和温阳益气活血方低、中、高剂量组,每组14只,另设空白组15只,给药组分别予相应药物灌胃,空白组和模型组予等量生理盐水,连续6周。超声检测大鼠心功能,ELISA检测血清脑钠肽(BNP)含量,计算心脏质量指数(HMI),HE染色观察心肌组织病理变化,酶活比色法、免疫组化及RT-PCR检测心肌组织Na^(+)-K^(+)-ATP酶、Ca^(2+)-ATP酶活性及表达。结果与空白组比较,模型组大鼠左室射血分数(LVEF)、左室短轴缩短率(LVFS)显著降低(P<0.01);血清BNP含量显著增加,HMI显著升高(P<0.01);心肌组织大量纤维水肿,胞质疏松,组织边缘心肌纤维溶解,伴有少量淋巴细胞浸润;心肌组织Na^(+)-K^(+)-ATP酶、Ca^(2+)-ATP酶活性显著减弱,蛋白及mRNA表达显著降低(P<0.01)。与模型组比较,温阳益气活血方高剂量组大鼠LVEF、LVFS显著升高(P<0.01);血清BNP含量显著减少,HMI显著降低(P<0.05,P<0.01);心肌纤维水肿、胞质疏松、炎症浸润、细胞空泡变性及纤维溶解程度均有改善;心肌组织Na^(+)-K^(+)-ATP酶和Ca^(2+)-ATP酶活性显著增强,蛋白及mRNA表达显著升高(P<0.01)。结论温阳益气活血方能通过调节心肌Na^(+)-K^(+)-ATP酶、Ca^(2+)-ATP酶活性及表达改善心力衰竭大鼠心功能不全,发挥治疗慢性心力衰竭的作用。

Objective To observe the effects of Wenyang Yiqi Huoxue Prescription on myocardial Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase in rats with chronic heart failure;To explore its mechanism of action in the treatment of heart failure.Methods A rat model of chronic heart failure was established by subcutaneous injection of isoproterenol at multiple points.The model rats were randomly divided into model group,Western medicine group,and Wenyang Yiqi Huoxue Prescription low-,medium-and high-dosage groups,with 14 rats in each group,and 15 rats in blank group.The administration groups were given corresponding drugs by gavage respectively,and the blank group and model group were given the same amount of normal saline for 6 consecutive weeks.Ultrasound was used to detect rat cardiac function,serum brain natriuretic peptide(BNP)content was detected by ELISA,heart mass index(HMI)was calculated,HE staining was used to observe the pathological changes of myocardial tissue,and Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase activity and expression in myocardial tissue were detected by enzyme activity colorimetry,immunohistochemistry and RT-PCR.Results Compared with the blank group,the left ventricular ejection fraction(LVEF)and left ventricular short-axis shortening rate(LVFS)of the rats in model group significantly decreased(P<0.01);serum BNP and HMI increased significantly(P<0.01);with a large number of fibrotic edema in the myocardial tissue,loose cytoplasm,myocardial fibrosis at the edge of the tissue,accompanied by a small amount of lymphocyte infiltration;the activities of Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase in myocardial tissue significantly decreased,and the protein and mRNA expressions significantly decreased(P<0.01).Compared with the model group,the LVEF and LVFS of the rats in Wenyang Yiqi Huoxue Prescription high-dosage group significantly increased(P<0.01);serum BNP and HMI decreased significantly(P<0.05,P<0.01);myocardial fibroedema,cytoplasmic looseness,inflammatory infiltration,cellular vacuolar degeneration and fibrinolysis were improved;the activities of Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase in myocardial tissue were significantly enhanced,and the protein and mRNA expressions significantly increased(P<0.01).Conclusion Wenyang Yiqi Huoxue Prescription can improve the cardiac insufficiency of rats with chronic heart failure by regulating the activity and expression of myocardial Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase,so as to play a role in the treatment of chronic heart failure.