基于外泌体中miR-9与ACE2-Ang（1-7）-MAS-ERK通路相关性探讨补肾降压方对原发性高血压大鼠心室重构的保护作用及机制

Hypertension has been one of the main diseases that are harmful to human health due to its high incidence and high mortality. Left ventricular hypertrophy (LVH) is an important target organ damage caused by hypertension, which is closely related to arrhythmia, sudden death and heart failure. Reversing ventricular hypertrophy is an important target for the treatment of hypertension. Exocrine RNA plays an important role in intercellular biological signals and functional material transfer in ventricular remodeling. MiR-9 in exosome is closely related to ventricular hypertrophy. ACE2-Ang (1-7) -MAS-ERK pathway in RAS related cell pathways is closely related to ventricular remodeling. There is definite evidence that exocrine is involved in the effect of RAS on ventricular remodeling. It is a hot research in exocrine related RNA and RAS cell pathway currently. Our previous studies showed that Bushen Jiangya decoction can stabilize blood pressure and reverse LVH, and its mechanism is related to up regulating microRNA-1 level and inhibiting ERK signaling pathway and its key target genes over expression. Therefore, we speculate that the Bushen Jiangya decoction may reverse LVH by regulating the miR-9 in the exocrine and then activating the ACE2-Ang (1-7) -MAS-ERK pathway. By establishing the model of ventricular hypertrophy in spontaneously hypertensive rats, we intend to reveal the molecular mechanism of Bushen Jiangya decoction on the left ventricular hypertrophy in spontaneously hypertensive rats, from the paracrine mechanism of tiny RNA regulating cell pathway in the exocrine body. This study can provide a scientific basis for the treatment of hypertension by the method of kidney tonifying, and also provide new directions and new ideas for the treatment of hypertension in clinic.

原发性高血压是危害人类健康的主要疾病之一，左室肥厚（LVH）是高血压导致的重要靶器官损害。逆转心室肥厚是治疗高血压病的重要目标。外泌体miR-9与心室肥厚关系密切，ACE2-Ang（1-7）-MAS-ERK是影响心室重构重要RAS旁分泌通路。研究表明外泌体RNA参与RAS心室重构，将外泌体RNA引入RAS通路研究是热点领域。前期研究示：补肾降压方稳定血压、逆转LVH，机制与上调microRNA-1抑制ERK信号通路有关。由此推测补肾降压方可能通过调控外泌体中miR-9激活ACE2-Ang（1-7）-MAS-ERK通路达到逆转LVH作用。本课题通过建立SHR心室肥厚模型及心肌肥大细胞，以ACE2-Ang（1-7）-MAS-ERK通路为切入点，应用qRT-PCR、Western blot、miRNA干扰和过表达技术，体内外实验相结合，探讨补肾法治疗LVH机制，为高血压病治疗提供新思路和靶标。

原发性高血压由于其高发病率和高致死率已成为危害人类健康的主要疾病之一。心肌肥厚是高血压性导致的重要靶器官损害，大量研究表明，左心室肥厚（Left ventricular hypertrophy，LVH）已成为心脑血管独立事件的危险因素，与心律失常、猝死、心力衰竭等密切相关。逆转心室肥厚是治疗高血压病的重要目标。研究表明：外泌体RNA在心室重构的细胞间生物信号及功能物质传递中发挥重要作用。我们的前期研究显示，基于补肾治法的补肾降压方能稳定血压、逆转LVH。因此，我们推测补肾降压方可能是通过调控外泌体中miR-9进而激活ACE2-Ang（1-7）-MAS-ERK通路而达到逆转LVH作用，认为该假说符合逻辑，科学合理。通过研究，我们得出结论：1.补肾降压方能够在SHR模型上稳定血压，减慢心率；2.补肾降压方能在一定程度上逆转心室肥厚；3.补肾降压方能够调节血浆外泌体中miR-9 miRNA表达；4.补肾降压方能够调控SHR大鼠血浆和组织中Ang II、Ang(1-7)浓度水平；5.补肾降压方能够下调SHR大鼠ACE mRNA，上调ACE2 mRNA及相关蛋白表达；6.补肾降压方逆转高血压心室肥厚的作用机制可能与调节血浆外泌体miR-9 miRNA进而激活ACE2-Ang（1-7）-MAS-ERK通路有关。通过体内和体外实验，观察补肾降压方对高血压左室肥厚的影响，从外泌体与相关通路探讨补肾降压方对高血压心室肥厚的可能机制，从而为补肾法治疗高血压提供科学依据，也为临床上寻找治疗高血压病新的治疗方法和新的药物靶点提供新思路。通过深层次揭示高血压左室肥厚的分子机制，阐明中医药治疗的科学内涵。