基于破骨细胞抑制的大血藤防治类风湿关节炎骨破坏物质基础及调控RANKL/RANK/OPG通路机制研究

Early prevention and treatment of bone destruction induced by rheumatoid arthritis can prevent secondary osteoporosis, osteoclasts play an important role in bone destruction of the pathological mechanisms. TCM treatment of rheumatoid arthritis has rich clinical experience. Sargentodoxae Caulis is one of the usual clinical traditional Chinese medicinal herbs for the treatment of fengshibitong and diepuzhongtong with TCM theory. We found that Sargentodoxae Caulis can inhibit osteoclast differentiation and function, promote osteoblast proliferation, inhibit inflammatory response in vitro, and reduce the expression level of RANKL in the serum of CIA rats with collagenⅡ-induced. According to the previous studies, TCM theory and clinical application, based on the inhibition of osteoclast differentiation and function, we will analyze the pharmacodynamic fingerprint of Sargentodoxae Caulis. Then we will combine the serum pharmacology with the serum pharmacochemistry, based on the strategy of target constituent “knock-out”, evaluate the effects by using CIA rats with kidney deficiency and co-culture model of synovial fibroblasts and monocytes. Our studies will define the effective substances for prevention and treatment of bone destruction in rheumatoid arthritis of Sargentodoxae Caulis, which can inhibit differentiation and function of osteoclasts. We will reveal the regulatory mechanisms of RANKL/RANK/OPG pathway. These results will provide the theoretical basis for the quality control and clinical application of Sargentodoxae Caulis.

早期防治类风湿关节炎患者骨破坏可预防继发性骨质疏松，破骨细胞在骨破坏病理机制中发挥重要作用。中医中药治疗类风湿关节炎具有丰富的临床经验，大血藤是中医和少数民族临床治疗风湿痹痛、跌扑肿痛的常用藤类中药民族药之一，申请者前期研究发现大血藤能抑制破骨细胞分化及功能，促进成骨细胞增殖，抑制炎症细胞的炎症反应，降低Ⅱ型胶原诱导关节炎大鼠血清RANKL水平。本项目拟在前期研究基础上，从中医药理论和临床应用出发，基于破骨细胞分化和功能抑制，开展大血藤谱效相关性分析，将中药血清药理学和中药血清药物化学方法相结合，基于目标成分敲除，应用体内病证结合的肾虚CIA大鼠及其滑膜成纤维细胞-单核细胞共培养诱生的破骨细胞模型进行药效评价，明确大血藤通过抑制破骨细胞分化和功能防治类风湿关节炎骨破坏的物质基础，揭示其调控RANKL/RANK/OPG通路的作用机制，为其质量控制和临床应用提供理论依据。

早期防治类风湿关节炎患者骨破坏可预防继发性骨质疏松，破骨细胞在骨破坏病理机制中发挥重要作用。中医中药治疗类风湿关节炎具有丰富的临床经验，大血藤是中医和少数民族临床治疗风湿痹痛、跌扑肿痛的常用藤类中药民族药之一。. 本项目在前期研究基础上，从中医药理论和临床应用出发，基于破骨细胞分化和功能抑制，建立并优化了诱导分化形成的破骨细胞模型、炎性骨破坏细胞模型、成骨细胞-破骨细胞共培养体系，以及病证结合的大鼠肾虚CIA模型；建立了大血藤指纹图谱，开展了大血藤与炎性骨破坏的谱效相关性分析，并辨识分析了主要成分；分离富集纯化了大血藤抑制破骨细胞分化和功能的成分（群），并验证了对类风湿关节炎骨破坏的作用；开展了大血藤成分（群）的作用机制研究。. 目前研究结果表明，大血藤总酚为其抑制破骨细胞分化和功能防治类风湿关节炎骨破坏的主要成分群，其中(-)-表儿茶素、原花青素B2、绿原酸、儿茶素、羟基酪醇、香草酸为主要候选成分；大血藤总酚及(-)-表儿茶素可抑制IL-1β、IL-6等炎症因子及肿瘤坏死因子TNF-α释放，抑制NF-κB/p65和p38 MAPK磷酸化，抑制破骨细胞分化相关转录因子或标志性基因TRAF6、TRAP、CTSK、NFATc1、c-fos表达，进而通过调控RANKL/RANK/OPG通路抑制破骨细胞的分化及骨吸收功能，发挥对骨破坏的抑制作用。. 本项目的研究，在一定程度上揭示了大血藤通过抑制破骨细胞分化和功能防治类风湿关节炎骨破坏的物质基础及其作用机制，为其质量控制和临床应用提供了理论依据。