miR-424-5p/miR-582-5p调控阳虚血瘀质膝骨关节炎的易感机制及补肾活血方干预作用研究

The pathogenesis of knee osteoarthritis (KOA) and its prevention and treatment with Traditional Chinese Medicine (TCM) have been the key research area our team focused on for years. Previously, we unconvered the signature of clinical epidemiology and preliminary mechanism of KOA. The preliminary data demonstrates: 1. More than 1/3 of KOA patients manifest Yang deficiency and blood stasis syndrome; 2. Metabolic disorder of chondrocytes is the main pathological manifestation; 3. The miR-424-5p/miR-582-5p expression is downregulated in chondrocytes belongs to Yang deficiency and blood stasis syndrome patients. Based on the findings above, we put forward the following hypothesis: Yang deficiency and blood stasis syndrome leads to miRNA abnormity, which is the key step to high susceptibility of KOA. Bushenhuoxue recipe reduces the susceptibility by regulating miR-424-5p/miR-582-5p, contributing to postponing or stopping the pathological process of KOA.This project intends to probe into the correlation between miRNA regulation and the micro-mechanism of TCM constitution, and further reveal the pathogenesis of KOA through the overexpression of miRNA and the technique of knockdown. Besides, through the Yang-warming and blood-activating therapy, can we intervene the homeostatic disorder of constitution of KOA patients. Thus we can prevent the occurrence and development of KOA by improving the constitution. The project is in line with the important strategic thought of "healthy China", through the implementation of the project, practice the concept of "cure the disease" and promote the development of TCM.

膝骨关节炎（KOA）的发生机制和中医药防治是课题组多年的研究方向。项目组前期完成了KOA临床流行病学调研与部分机制研究，结果表明：1.多于三分之一KOA患者属于阳虚血瘀质；2.软骨细胞代谢紊乱是其主要病理表现；3.阳虚血瘀质KOA中软骨细胞的miR-424-5p/miR-582-5p表达降低。基于上述发现，提出科学假说：阳虚血瘀质的miRNA异常是导致高KOA易感性的关键环节，补肾活血方通过调控miR-424-5p和miR-582-5p降低其易感性，进而延缓或阻断KOA的病理进程。本课题拟通过体内外miRNA的过表达和敲降技术，深入探析miRNA调控与中医体质微观机制之间的相关性，进一步揭示KOA发病机制。同时应用补肾活血方干预KOA体质的偏颇状态，实现调质防病和调质防变。课题符合“健康中国”重大战略思想，通过课题的实施，践行“治未病”理念、推进中医事业发展。

膝骨关节炎（Knee Osteoarthritis，KOA）是一种中老年人常见多发的以软骨退行性变和继发骨质增生为主，最终发生膝关节功能障碍的一种退行性疾病。中医体质学以中医理论为主导，分析疾病的反应状态、病变的性质和发展趋向，指导预防和治疗的学说。KOA的发生机制和中医药防治是本项目团队多年的研究方向。项目组前期的KOA临床流行病学调研与部分机制研究，发现多于三分之一KOA患者属于阳虚血瘀质，并且这些患者miRNA 表达异常。同时，项目组的补肾活血方临床上缓解阳虚血瘀质KOA关节疼痛，改善关节功能。因此，本项目从中医体质学角度研究阳虚血瘀质KOA人群的遗传易感因素和机制。利用原代软骨细胞和阳虚血瘀质动物模型，分析miRNA表达，项目组发现阳虚血瘀质患者的hsa-miR-582-5p低表达是导致KOA高度易感的关键环节。通过荧光素酶报告实验和病毒介导的体内敲低技术，证明hsa-miR-582-5p通过靶向抑制Adamts5表达，促进软骨细胞外基质合成。补肾活血方通过上调miRNA表达，降低了阳虚血瘀质KOA模型的软骨退变和软骨下骨硬化，进而延缓KOA的病理进程。本项目深入探析了miRNA调控与中医体质微观机制之间的相关性，进一步揭示KOA发病机制。同时应用补肾活血方干预KOA体质的偏颇状态，实现调质防病和调质防变。课题符合“健康中国”重大战略思想，通过课题的实施，践行治未病”理念、推进中医事业发展。