基于TRPV4/SOX9/RUNX2信号轴探索牛蒡子汤对超负荷诱导早中期KOA干预的机制研究

Knee osteoarthritis (KOA) is a chronic osteoarthritis characterized by the degeneration of articular cartilage. The pathogenesis is mainly related to biomechanical imbalance and inflammatory response, and overloaded mechanical stimulation resulting in chondrocyte proliferation, dysfunction is an important factor in accelerating cartilage degeneration. The classic Niubangzi decoction of Shi’s traumatology has significant effect on KOA prevention and treatment. The preliminary research confirmed that the arctigeninc has the function of targeting activation mechanosensitive ion channel TRPV4, enhancing the production of SOX9 and promoting the proliferation and differentiation of chondrocytes. However, it has not been reported whether it acts through the TRPV4 / SOX9 / RUNX2 signal axis. Our group speculates that Niubangzi decoction and the active ingredients targeting enhance the expression of transcription factor SOX9 through the activation of TRPV4 pathway and then inhibit the transcription factor RUNX2 expression to enhance chondrocyte proliferation and differentiation. This project will clarify the mechanism of the effect of Niubangzi decoction and its active ingredients on TRPV4 / SOX9 / RUNX2 signal axis from mechanics point of view. This project will explore the mechanism of TRPV4 / SOX9 / RUNX2 signal axis by constructing cell and animal models under different mechanics and elucidate the mechanism of Niubangzi decoction and its active ingredients against KOA from the mechanical point of view.

膝骨关节炎(knee osteoarthritis, KOA)是以关节软骨退变为特征的慢性骨关节病，其发病机制主要与生物力学失衡和炎症反应有关，而超负荷力学刺激导致的软骨细胞增殖与分化功能失调是加速软骨退变的重要因素。石氏伤科经典方牛蒡子汤防治KOA临床疗效显著，前期已证实其君药提取物牛蒡子苷元具有靶向激活力学敏感通路TRPV4，增强SOX9生成和调节软骨细胞增殖与分化的功能，但是否通过TRPV4/SOX9/RUNX2信号轴作用未见报道。本课题组推测牛蒡子汤及有效成分是通过激活TRPV4通路继而靶向增强转录因子SOX9表达进而抑制转录因子RUNX2表达来增强软骨细胞增殖与分化。本课题将通过构建不同力学作用下的细胞和动物模型，探寻TRPV4/SOX9/RUNX2信号轴在调节软骨细胞生物学功能中的机制，从力学角度阐明牛蒡子汤及有效成分防治KOA作用的机理。

膝骨关节炎（KOA）是中老年人常见疾病之一，具有较高的致残率，给家庭和社会带来沉重的负担，学界普遍认为KOA的发生是多因素综合作用的结果，且膝关节生物力学改变对KOA的发生、发展具有较大的影响。基于石氏伤科经典方牛蒡子汤防治KOA临床疗效显著，及前期研究已证实牛蒡子苷元具有靶向激活力学敏感通路TRPV4，增强SOX9生成和调节软骨细胞增殖与分化的功能。.本课题从力学角度探讨牛蒡子汤及有效成分调控TRPV4/SOX9/RUNX2信号轴，以阐明其增强软骨细胞增殖与分化，调节软骨细胞凋亡与基质降解改善软骨退化的作用机制。主要研究内容：动物层面，先建立超负荷应力损伤KOA小鼠动物模型，继用牛蒡子汤和龙胆苦苷分别对超负荷诱导早中期KOA小鼠进行干预，观察疗效。在细胞层面，先制备牛蒡子汤冻干粉，干预软骨细胞，观察其调控TRPV4/SOX9/RUNX2信号轴，对软骨细胞增殖与分化的影响；并基于TRPV4/SOX9/RUNX2信号轴探索牛蒡子苷元对软骨细胞的保护作用及机制。运用多组学技术分析模拟步行压力对软骨的干预作用及机制。基于多组学得出的压力关键基因（蛋白），验证压力下牛蒡子苷元对软骨的保护机制。探讨超负荷应力作用对软骨细胞凋亡与基质降解的调控作用。在超负荷应力作用下，探讨TRPV4通过ERK信号通路调控软骨细胞凋亡与基质降解的作用机制。探讨龙胆苦苷通过TRPV4/ERK信号轴调控软骨细胞凋亡与基质降解的分子机制研究。.综上研究，通过构建不同力学作用下的细胞和动物模型，利用分子生物学、生物信息学等技术手段，从多途径、多层次、多靶点证实了力学对KOA发生发展的影响，明确了牛蒡子汤及其有效成分对软骨增殖与分化的作用，初步证实了牛蒡子汤及其有效成分调控软骨细胞TRPV4/SOX9/RUNX2信号轴表达从而防治KOA的实验依据，揭示了该复方制剂及单体防治KOA的治疗机制，为今后牛蒡子汤及其有效成分治疗KOA的转化研究提供了理论依据。