补肾通络法介导OPG／RANKL／RANK系统调控膝骨性关节炎软骨及软骨下骨代谢的机制研究

Knee osteoarthritis is the most common degenerative bone disease in humans, and also become a hot spot in global public health. Cartilage and subchondral bone are the basic structural and functional units of the knee joint. Studies have shown that the OPG/RANKL/RANK system plays an important role in the regulation of cartilage and subchondral bone metabolism. It may be a target for the treatment of osteoarthritis. .According to the theory of traditional Chinese medicine, such as "kidney bone" and "meridians of obstructions", setting up Rat model of knee osteoarthritis induced by double forelimbs and a monoclonal cell line carrying luciferase reporter gene as study subjects, Using methods such as cell and molecular biology. To study the mechanism of Bushen Tongluo traditional Chinese medicine mediating OPG/RANKL/RANK system on the regulation of cartilage and subchondral bone metabolism. To further lay the foundation for the treatment of degenerative bone disease with "Bushen Tongluo" and its prescriptions. In order to make the In-depth exploration of the modern biological characteristics of the pathogenesis theory of "long-term injury, injury to kidney and meridians, and obstruction of the kidneys", revealing the Internal Law of TCM Pathogenesis Theory of "Kidney Bone and Meridian Obstruction", establishing the Theory and Method of Modern Biology Research on the Theory of Bushen Tongluo, enriching and developing the scientific connotation of the theories of “kidney bone, Meridian Obstruction” and “Kidney Clearance Method”.

膝关节骨性关节炎(Knee Osteoarthritis)是人类最常见的退变性骨病，已成为全球公共卫生关注的热点。软骨及软骨下骨是膝关节的基本结构和功能单位。研究表明OPG／RANKL／RANK系统在调节软骨及软骨下骨代谢方面具有重要作用，将可能成为骨关节炎治疗的靶点。.根据“肾主骨”和“经脉痹阻”等中医病机理论，以去双前肢诱导直立大鼠膝骨性关节炎模型和携带荧光素酶的报导基因单克隆细胞株为研究对象，采用细胞和分子生物学等方法，研究补肾通络中药介导OPG／RANKL／RANK系统调控软骨及软骨下骨代谢的机制，进一步奠定“补肾通络法”及其方药治疗骨退变性疾病的基础，深入探讨“久立伤骨损于肾及经脉痹阻”病机理论的现代生物学特性，揭示“肾主骨及经脉痹阻”中医病机理论的内在规律，建立“补肾通络法”理论现代生物学研究思路和方法，丰富和发展中医“肾主骨、经脉痹阻”和“补肾通络法”理论的科学内涵。

骨关节炎是人类最常见的退行性骨病，已成为全球公共卫生关注的热点。软骨及软骨下骨是膝关节的基本结构和功能单位。研究表明OPG／RANKL／RANK系统在调节软骨及软骨下骨代谢方面具有重要作用，已成为骨关节炎治疗的潜在靶点。本课题根据“肾主骨”和“经脉痹阻”中医学藏象和病机理论，以去双前肢诱导直立大鼠膝骨关节炎模型为研究对象，采用分子生物学等方法，研究补肾通络中药介导OPG／RANKL／RANK系统调控软骨及软骨下骨代谢的机制。成功建立了去双前肢直立、去双前肢直立联合假Hulth、去双前肢直立联合Hulth三种大鼠模型，其中去双前肢直立联合Hulth模型组中的膝关节退变更为典型。去双前肢直立联合Hulth组中大鼠膝关节软骨Mankin评分最高，CollagenⅡ阳性细胞比例最低，MMP-13阳性细胞比例最高，大鼠膝关节软骨的退变最严重。骨痹合剂组中大鼠膝关节软骨Mankin评分最低，CollagenⅡ阳性细胞比例最高，MMP-13阳性细胞比例最低，膝关节退变较轻。骨痹合剂对去双前肢直立联合 Hulth 大鼠膝关节炎模型软骨退变起到延缓作用，具体通过上调TIMP-1表达、下调 VEGF表达，延缓了膝骨关节炎模型软骨的病变进程。骨痹合剂对去双前肢直立联合 Hulth 法大鼠膝骨关节炎模型产生了抗炎机制，可通过下调IL-1β和TNF-α的表达，延缓了膝骨关节炎软骨细胞的退变进程。骨痹合剂可降低软骨下骨中IL-1β mRNA、IL-6 mRNA的表达，减轻炎症反应，下调软骨下骨中RANK mRNA，并上调OPG mRNA、ALP mRNA、COL-Ⅰ mRNA的表达，通过调节骨代谢状态并促进软骨下骨的修复。Western blot法检测结果提示骨痹合剂可通过下调RANK的蛋白表达，上调OPG、COL-Ⅰ的蛋白表达，达到良性调节软骨下骨代谢，延缓膝骨关节炎软骨下骨退变进程。本课题阐述了“补肾通络法”及其方药治疗骨退变性疾病的基础，深入探讨了“久立伤骨损于肾”及“经脉痹阻”中医藏象和病机理论的现代生物学特性，揭示了“肾主骨及经脉痹阻”中医藏象和病机理论的内在规律，完善了“补肾通络法”理论的现代生物学研究思路和方法，丰富和发展了中医“肾主骨、经脉痹阻”和“补肾通络法”理论的现代科学内涵。