益气活血法对舒张性心力衰竭气虚血瘀证大鼠心肌细胞Ca2+瞬变影响及补阳还五汤干预的实验研究

Diastolic heart failure（DHF） is one of the challenges in the field of cardiovascular research. There is no effective treatment for diastolic heart failure at present. Our research group intends to start with the pre-project experimental results that endothelial cells and myocardial cell Ca2+ transients in myocardial force and frequency relationship (FFR) play a vital role in myocardial diastolic function. Whether traditional Chinese medicine compound work on these targets which was mentioned has not yet reached to any conclusions now.Based on the previous investigation and research on the syndrome of diastolic heart failure in our hospital our research group propose the hypothesis that supplementing Qi and activating blood method can treat diastolic heart failure disease. We intend to treat rats with diastolic heart failure in using Buyanghuanwu decoction and Measure the rat myocardial diastolic function using the ultrasonic get the myocardial systolic and diastolic force FFR have a quantitative analysis of cardiac endothelial cells applying laser scanning confocal microscopy and the rat cardiomyocytes Ca2+ transients Determine the SERCA2a enzyme activity the expression of phosphoprotein and Ca2+ regulatory protein level. To explore the effect of Buyanghuanwu decoction on diastolic heart failure.There is important significance for this study to clarify Buyanghuanwu decoction which is good at supplementing Qi and activating blood works by affecting Ca2+ transient in myocardial cells repairing of myocardial endothelial cells and FFR affecting myocardial diastolic function.This study will lay the necessary theoretical foundation for classical prescription and supplementing Qi and activating method in the treatment of diastolic heart failure disease Qi deficiency and blood stasis syndrome and provides the beneficial exploration.

舒张性心力衰竭（DHF）是心血管研究领域的难题之一，尚无有效的治疗方法，课题组拟从前期预实验入手：即心肌细胞内Ca2+ 瞬变、心肌内皮完整性和心肌力与频率关系（FFR）对心脏舒张功能起着非常重要的作用。中药复方对以上靶点是否有作用尚无结论，在前期对DHF证候调查的基础上,提出应用益气活血法治疗DHF的假说，拟用补阳还五汤干预DHF气虚血瘀证大鼠，超声心动图测量大鼠心脏舒张功能，测定心肌活组织收缩力和舒张力、FFR，应用激光共聚焦显微镜对心肌内皮细胞定量分析，测定SERCA2a酶活性、检测受磷蛋白和Ca2+调节蛋白水平的表达，测量大鼠心肌细胞内Ca2+ 瞬变,观察补阳还五汤对DHF的疗效。本研究对阐明功在益气活血的补阳还五汤是否是通过影响心肌细胞内Ca2+ 瞬变、修复心肌内皮和FFR进而改善心肌舒张功能有重要的意义，为应用经典方药益气活血法治疗DHF气虚血瘀证奠定必要的基础提供有益的探索。

本项目研究益气活血法对舒张性心力衰竭大鼠的干预效应及钙瞬变的影响。在项目执行过程中，本项目课题组建立了舒张性心力衰竭大鼠模型，观察了益气活血法及补阳还五汤对舒张性心力衰竭大鼠超声心动图下二尖瓣环舒张早期运动速度/二尖瓣环舒张晚期运动速度（e’/a’）比值的影响；测量了心肌收缩力、舒张力，应用蛋白免疫印迹法（Western Blot）及聚合酶链式反应法（PCR）检测心肌Ca2+调节蛋白水平的表达；应用激光共聚焦显微镜测量大鼠心肌细胞内Ca2+瞬变。研究结果表明舒张性心力衰竭大鼠心肌肌浆网钙泵（sarcoplasmic reticulum Ca2+-ATPase2a，SERCA2a）和受磷蛋白（phospholamban，PLB）钙转运蛋白表达和mRNA的表达水平下降，而补阳还五汤可能是通过提高SERCA2a和PLB的蛋白表达和mRNA转录水平来调节舒张性心力衰竭大鼠左室的舒张功能，表明益气活血法代表方补阳还五汤可以在一定程度上改善舒张性心力衰竭大鼠左室舒张功能。同时，本研究认为补阳还五汤可能是通过影响心肌细胞内Ca2+ 瞬变,进而影响心肌的收缩和舒张功能。因此本项目，初步揭示了应用益气活血法对舒张性心力衰竭气虚血瘀证大鼠的作用并阐述其机制，为中医药治疗舒张性心力衰竭的研究奠定必要的基础，为从新角度研究中药复方治疗舒张性心力衰竭大鼠提供新的思路。