基于TGF-β/Smad通路探讨高血压心衰证候本质及心肾纤维化机制

Based on the previous research, this project try to pinpoint the syndrome essence and specific sensitive index of hypertensive heart failure rat model according to "syndrome detection by formula" method, by observing the intervention effect of heart and kidney fibrosis, the improvement of cardiac and renal function, especially the improvement of renal function after heart failure through the regulation of TGF-β/Smad signaling pathway after administering Shen-Fu injection and Shen-Mai injection on Dah1 salt sensitive rat model of heart failure,. It would lay a solid foundation for construction of combined disease and syndrome rat model of heart dual-deficiency of qi and yin syndrome of hypertensive heart failure, and it also would provide the experimental basis for the reasonable application of traditional Chinese medicine preparation in hypertensive heart failure by exploring the indications and contraindications of traditional Chinese medicine injection in hypertensive heart failure; Combined experiments in vivo and in vitro, we try to explore the pathological mechanism and the key target of heart and kidney fibrosis of hypertensive heart failure rat model, by using ELISA, Western-blot, RT-PCR, flow cytometry, immunofluorescence, laser scanning confocal and electron microscopy technology, from the protein level to gene level. It would provide the pathological evidence of heart kidney interaction theory and clarify that the heart and kidney bidirectional damage is a pathological link of hypertensive heart failure， the heart and renal fibrosis is the key pathogenesis of the occurrence and development of hypertensive heart failure and all of these would have very important significance.

本项目是在前期研究工作的基础上，基于“以方测证”理论，运用Dahl盐敏感大鼠复制高血压心衰模型，通过参脉注射液与参附注射液，调控TGF-β/Smad信号通路、干预心肾纤维化、改善大鼠心、肾功能的作用，特别是改善心衰后肾功能的效果，明确高血压心衰大鼠模型的证候本质与特异性敏感指标，为构建高血压心衰心气阴虚证病证结合模型打下坚实的基础，探讨中药注射剂对高血压心衰的适应证和禁忌证，为高血压心衰中药制剂的合理应用提供实验依据；结合在体实验和体外细胞实验，运用ELISA、Western-blot、RT-PCR、流式细胞术、免疫荧光、激光共聚焦、电子显微镜等技术，从蛋白水平到基因水平，探讨高血压心衰大鼠心、肾纤维化的病理机制和关键作用靶点，为“心肾相交”理论提供现代病理依据，阐明心肾双向损害是高血压心衰发生发展的病理环节，而心肾纤维化是高血压心衰发生发展的关键病理机制，具有非常重要的意义。

心衰为多种心血管疾病(高血压病、心肌梗死等)的终末阶段，其致死率高、并发症多，是我国严重的公共卫生问题。心肌纤维化是高血压心衰发展的关键病理环节，其严重程度是决定高血压心、肾两脏靶器官损害发展及预后的主要因素。TGF-β是促纤维化发展的关键生长因子，通过TGF-β/Smad信号通路调节Smad2/3活化，参与心肌纤维化的发展。故本研究以盐敏感大鼠为研究对象，以经典TGF-β/Smad通路为切入点，通过高盐饮食构建高血压心衰大鼠模型，以参麦、参附注射液进行干预，结合“以方测证”阐明其中医证候本质，指导临床用药，并进一步从动物实验、细胞实验阐明高血压心衰心肌纤维化的发病机制及参麦注射液治疗靶点，为心衰的防治提供理论依据。.本研究发现，高血压心衰大鼠心肌、肾组织 TGF-β/Smad 信号转导通路活化，并参与肾小管上皮-间质细胞转分化过程，证实了心肾纤维化是高血压心衰发生发展的关键环节，TGF-β/Smad 信号通路活化是其关键病理机制之一。参麦注射液通过调控 TGF-β/Smad 信号转导通路而抑制心、肾纤维化状态，其用机制与抑制 TGF-β1/Smad信号转导通路活化及肾小管上皮-间质细胞转分化过程有关。综上所述，本研究基于“以方测证”确证了高血压心衰大鼠为“心气阴虚证”，并揭示了高血压心衰大鼠心肾肌纤维化的病理机制及参麦注射液治疗高血压心衰的关键作用机理，为心衰临床诊治及中药注射液的应用提供实验指导。.本课题共发表“期刊论文34篇 + 学位论文6篇”，其中Sci论文 5 篇，CSCD论文12 篇，其他论文17 篇；获中国中西医结合学会科学技术奖三等奖1项，湖南省中医药科技奖一等奖1项；培养博士研究生2名，硕士研究生4名，获湖南省优秀硕士学位论文1篇，获国家研究生奖学金3名、湖南省优秀硕士毕业生1名。