补肾活血方对心梗后心衰大鼠心室重构及心脑组织AVP、AQPs表达的影响

Effect of Bushen Huoxue Prescription on Ventricular Remodeling and Expression of AVP and AQPs in Heart and Brain of Rat Model of Heart Failure after Myocardial Infarction

摘要:

目的 探讨补肾活血方对心肌梗死后心力衰竭大鼠脑组织精氨酸加压素（AVP）、水通道蛋白（AQP1、AQP4）及心肌水通道蛋白（AQP1、AQP4、AQP7）表达的影响。方法 选取健康雄性SD大鼠60只，随机分为假手术组、模型组、补肾活血组与托伐普坦组，假手术组大鼠仅开胸冠状动脉（冠脉）挂线不结扎，其他各组采用结扎左冠状动脉前降支联合力竭式游泳、饥饿建立心肌梗死后心力衰竭模型。模型评价成功后，托伐普坦组与补肾活血组分别采用托伐普坦（1.35 mg·kg^-1）及补肾活血方（15.75 g·kg^-1）进行干预，假手术组和模型组采用等体积生理盐水干预，持续4周。彩色多普勒心动超声测定大鼠左心室结构及功能变化，酶联免疫吸附测定法（ELISA）检测血清N末端B型利钠肽前体（NT-proBNP）含量，苏木素-伊红（HE）染色观察大鼠脑组织、心肌组织形态，蛋白免疫印迹法（Western blot）检测大鼠脑组织AVP、AQP1、AQP4和心肌组织AQP1、AQP4、AQP7蛋白表达变化，实时荧光定量聚合酶链式反应（Real-time PCR）检测脑组织AVP、AQP1、AQP4 mRNA表达水平，检测心肌组织AQP1、AQP4、AQP7 mRNA表达水平。结果 与假手术组比较，模型组大鼠左心室内径（LVID）显著增加（P<0.01）、射血分数（EF）、短轴缩短率（FS）显著减少（P<0.01）、左心室室壁厚度（LVS、LVPW）显著增厚（P<0.01）；血清NT-proBNP含量显著增加（P<0.01）；大鼠脑组织神经细胞水肿、空泡样变，大鼠心肌梗死区心肌细胞坏死，心肌纤维断裂；脑组织AVP、AQP1、AQP4 mRNA及蛋白表达显著升高（P<0.01），心肌组织AQP1、AQP4、AQP7 mRNA及蛋白表达显著升高（P<0.01）。与模型组比较，托伐普坦组和补肾活血组大鼠左心室内径（LVID）、射血分数（EF）、短轴缩短率（FS）、左心室室壁厚度（LVS、LVPW）均有不同程度改善（P<0.05，P<0.01）；血清NT-proBNP含量明显减少（P<0.05）；脑组织神经细胞水肿减轻，心肌纤维排列较整齐，部分心肌细胞坏死；脑组织AVP、AQP1、AQP4 mRNA及蛋白表达显著降低（P<0.01），心肌组织AQP1、AQP4、AQP7 mRNA及蛋白表达均不同程度降低（P<0.05，P<0.01）。结论 补肾活血方能通过下调心肌梗死后心力衰竭大鼠脑组织AVP、AQP1、AQP4表达，下调心肌组织AQP1、AQP4、AQP7表达，来调节心脑水液代谢障碍，延缓心室重构，改善心功能。

Abstract:

Objective To explore the effects of Bushen Huoxue prescription on the expression of arginine vasopressin （AVP） and aquaporins （AQP）1 and AQP4 in the brain and AQP1， AQP4， and AQP7 in the myocardial tissue of the rat model of heart failure after myocardial infarction.Method Sixty healthy male SD rats were selected and randomized into sham surgery， model， Bushen Huoxue prescription， and tolvaptan groups. The rats in the sham surgery group only had their coronary arteries exposed without ligation， while those in the other groups were subjected to ligation of the left coronary artery combined with exhaustive swimming and starvation for the modeling of heart failure after myocardial infarction. After the successful modeling， the rats in the tolvaptan （1.35 mg·kg^-1） and Bushen Huoxue prescription （15.75 g·kg^-1） groups were treated with corresponding drugs， and those in the sham surgery and model groups were treated with an equal volume of normal saline for 4 weeks. Color doppler echocardiography was employed to measure changes of the left ventricular structure and function in rats. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the serum level of N-terminal pro-B-type natriuretic peptide （NT-proBNP）. Hematoxylin-eosin （HE） staining was conducted to reveal the pathological changes in the myocardial tissue. Western blotting was employed to measure the protein levels of AVP， AQP1， and AQP4 in the brain tissue and AQP1， AQP4， and AQP7 in the myocardial tissue. Real-time fluorescence quantitative PCR was conducted to determine the mRNA levels of AVP， AQP1， and AQP4 in the brain tissue and AQP1， AQP4， and AQP7 in the myocardial tissue.Result Compared with the sham surgery group， the model group showed increased left ventricular internal diameter （LVID） （P<0.01）， decreased ejection fraction （EF） and short axis fractional shortening rate （FS） （P<0.01）， increased left ventricular septal and posterior wall thicknesses （LVS and LVPW） （P<0.01）， elevated serum NT-proBNP level （P<0.01）， edema and vacuolar degeneration of nerve cells in the brain tissue， necrosis of myocardial cells and rupture of myocardial fibers in the infarcted area， and upregulated mRNA and protein levels of AVP， AQP1， and AQP4 in the brain tissue （P<0.01） and AQP1， AQP4， and AQP7 in the myocardial tissue （P<0.01）. Compared with the model group， tolvaptan and Bushen Huoxue prescription restored LVID， EF， FS， LVS， and LVPW （P<0.05， P<0.01）， lowered the serum NT-proBNP level （P<0.05）， alleviated the edema of nerve cells in the brain tissue， improved the arrangement of myocardial fibers， mitigated the necrosis of myocardial cells， and downregulated the mRNA and protein levels of AVP， AQP1， and AQP4 in the brain tissue （P<0.01） and AQP1， AQP4， and AQP7 in the myocardial tissue （P<0.05， P<0.01）.Conclusion Bushen Huoxue prescription can regulate cardiovascular and cerebrovascular fluid metabolism disorders， delay ventricular remodeling， and improve the cardiac function by downregulating the expression of AVP， AQP1， and AQP4 in the brain tissue and AQP1， AQP4， and AQP7 in the myocardial tissue in the rat model of heart failure after myocardial infarction.