基于MiR-155-5p/eNOS通路探究杜仲刺蒺藜组分药对拮抗高血压肾损害的作用机制

Hypertensive kidney damage is an important cause of chronic kidney disease. Improving glomerular endothelial function and eNOS activity can effectively reduce the incidence of hypertensive renal damage. MiR-155-5p is an important regulator to increase eNOS activity. The project takes the disease identification of traditional Chinese medicine and the theory of Chinese herbology for the guidance, as well as linking up modern medical theory. Based on the protection of hypertensive target organ, we take the hypertension kidney damage as the entry point and from the perspective of epigenetics, the high-throughput sequencing technology will be used to construct the MicRNA expression profile. Through the establishment of the external injury model, the target MicRNA of regulating eNOS of double-drugs of Eucommia & Tribulus terrestris will be determined. MiR-155-5p is overexpressed and knocked-down in vitro and in vivo respectively, and we will explore the mechanism of double-drugs of Eucommia & Tribulus terrestris in increasing the NO content and eNOS activity through controlling of the MiR-155-5p. The project aims to deeply explore the multi-pathway and multi-target mechanism of anti-hypertensive renal damage, and provide a basis for the protection of the target organ of hypertension by traditional Chinese medicine.

高血压肾损害是是慢性肾脏病的重要原因，改善肾小球内皮功能，提高eNOS活性能够有效降低高血压肾损害的发病率，MiR-155-5p是提高eNOS活性的重要调节分子。本项目以中医辨病和中药药对组方理论为指导，衔接现代医学理论，以高血压肾损害为切入点，立足于高血压靶器官保护，从表观遗传学的视角，应用高通量测序技术构建MicRNA表达谱，通过体外损伤模型的建立，明确杜仲刺蒺藜组分药对调控eNOS拮抗肾小球内皮细胞氧化损伤的靶MicRNA，体外、体内分别过表达及敲减MiR-155-5p，探晰杜仲刺蒺藜组分药对靶向调控MiR-155-5p以提高NO含量、eNOS活性的效应机制，以期深入解读中药多通路多靶点抗高血压肾损害的机制，为中医药保护高血压靶器官提供依据。

高血压病是世界性的重大公共卫生问题，严重危害和威胁着人类的健康。高血压肾损害是严重的高血压并发症，是慢性肾脏病的重要原因。研究表明, 改善肾小球内皮功能，提高eNOS活性能够有效降低高血压肾损害的发病率，miR-155-5p是提高eNOS的重要调节因子。本项目以中医辨病和中药药对组方理论为指导，衔接现代医学理论，以高血压肾损害为切入点，立足于高血压靶器官保护，从基因学的视角，应用高通量测序技术构建MicRNA表达谱，为深入解读中药多通路多靶点抗高血压肾损害的机制，为中医药保护高血压靶器官提供依据。本项目采用体外实验的办法，培养WKY大鼠肾小球内皮细胞，建立AngⅡ损伤模型，观察经杜仲刺蒺藜组分药对干预后肾组织NO含量，eNOS表达等变化，明确杜仲刺蒺藜组分药对调控肾小球内皮细胞eNOS损伤机制，利用高通量测序技术筛选鉴定MicRNA序列表达谱，明确杜仲刺蒺藜组分药对的有效MicRNA靶点；采用体外实验的办法，对肾小球内皮细胞MicRNA155-5p进行过表达，观察经杜仲刺蒺藜组分药对干预后肾组织NO含量，eNOS表达等变化，明确了杜仲刺蒺藜组分药对对靶向调控MicRNA155-5p对eNOS表达的作用，阐释其调控的内在通路和作用靶点；采用体内实验的办法，对SHR大鼠MicRNA155-5p进行过表达，观察经杜仲刺蒺藜组分药对干预后大鼠肾脏，主动脉，NO含量，eNOS表达的变化，验证杜仲刺蒺藜对靶向调控MicRNA155-5p的作用，揭示杜仲刺蒺藜对拮抗eNOS的作用机制。本项目从高血压肾小球内皮细胞损伤的视角，明确抗高血压肾损害中药杜仲刺蒺藜组分药对的有效靶点MiR-155-5p，揭示组分药对靶向干预MiR-155-5p调控eNOS活性、NO含量拮抗肾小球内皮细胞氧化损伤的保护机制，揭示了中医药多通路、多靶点的科学内涵本质，具有重要的探索价值。本研究共计申请专利1项，发表SCI文章6篇，中文文章7篇，其中北大中文核心2篇。