补肾活血方调控Wnt/β-catenin和TGF-β双通路干预骨关节炎的分子机制研究

Osteoarthritis (OA) is the most common bone metabolic disease, correlated to Wnt/β-catenin and TGF-β signaling pathway-mediated unbalance of bone metabolic. MMP-13 is the common end for those pathways. It is known that kidney-deficiency and blood stasis is the TCM pathogenesis of OA, which is suitable for Bu Shen Huo Xue treatment. Preliminarily, we made a Bu Shen Huo Xue formula and evidenced its anti-OA effect with MMP-13 inhibitory activity. Therefore we proposed a scientific hypothesis: the anti-OA mechanism of Bu Shen Huo Xue formula is related to the synergistic modulation of both Wnt/β-catenin and TGF-β signaling pathways. To verify this hypothesis, this study plans to establish a classic OA model as well as two transgenic OA model (activation of Wnt/β-catenin signaling and inhibition of TGF-β signaling) to demonstrated the synergistic effect of those pathways in OA and reveal the two-pathway based molecular mechanism of Bu Shen Huo Xue formula for OA treatment. Through serum pharmacochemistry and in vitro experiment, we can verify the action mechanism of Bu Shen Huo Xue formula from the three levels of “formula-herb-component”. This study will eventually clarify the pathogenesis of OA and action mechanism of Bu Shen Huo Xue formula, and also provide new ideas for anti-OA study on TCM formula.

骨性关节炎（OA）是最常见的骨代谢疾病，与Wnt/β-catenin和TGF-β信号通路共同介导的骨代谢失衡有关，MMP-13是两条通路的共同节点。已知肾虚血瘀是OA的中医病机，适用补肾活血法干预。项目组前期自拟补肾活血方，证实其能通过抑制软骨MMP-13活性对OA发挥干预作用，由此我们提出科学假说：补肾活血方有效干预OA的作用机制与Wnt/β-catenin和TGF-β双信号通路的协同调控有关。为验证该假说，本项目拟建立OA经典模型和转基因/基因敲除模型（Wnt/β-catenin通路活化和TGF-β通路阻断），从体内验证两条通路对OA的协同作用、揭示补肾活血方基于双通路调控干预OA的分子机制；再通过血清药物化学和细胞体外实验，从“方—药—成分”三个层次验证补肾活血方的调控机制。本项目研究将为最终阐明OA的发病机理以及补肾活血方的干预机制奠定基础，为中药复方治疗OA的研究提供新的思路。

骨性关节炎（OA）是最常见的骨代谢疾病，与Wnt/β-catenin和TGF-β信号通路共同介导的骨代谢失衡有关，MMP-13是两条通路的共同节点。已知肾虚血瘀是OA的中医病机，适用补肾活血法干预。项目组前期自拟补肾活血方，证实其能通过抑制软骨MMP-13活性对OA发挥干预作用，由此我们提出科学假说：补肾活血方有效干预OA的作用机制与Wnt/β-catenin和TGF-β双信号通路的协同调控有关。为验证该假说，本项目建立OA经典模型和转基因/基因敲除模型，探究补肾活血方基于双通路调控干预OA的分子机制；再通过和细胞体外实验，揭示补肾活血方的调控机制。研究初期发现，补肾活血方对Wnt/β-catenin转基因模型不敏感，故只采用TGF-β条件敲除OA模型进行机制探索。研究结果首先验证补肾活血方基于MMP-13调控干预大鼠OA模型的药效作用，明确其只对TGF-β信号通路有显著调控作用，但对Wnt/β-catenin信号通路作用不明显，该方能通过激活TGF-β信号通路抑制Mmp13改善OA软骨代谢、减缓退变进程，为OA的治疗提供了新的途径，为补肾活血方治疗OA的临床应用提供科学思路。