Tianhe Zhuifeng Gao reverses inflammatory response and attenuates bone/cartilage destruction in rheumatoid arthritis via PSMC2-RUNX2-COL1A1 axis based on transcriptional regulatory network analysis and experimental validation

<b>Background: </b>Tianhe Zhuifeng Gao (TZG) is an authorized Chinese patent drug with satisfying clinical efficacy, especially for RA patientswith cold-dampness syndrome. However, its underlyingpharmacological mechanisms remain unclear.<b>M</b><b>ethod</b><b>:</b><b> </b>Anti-arthritic effects of TZG were evaluated using an adjuvant-induced arthritis (AIA) ratmodel. Transcriptional regulatory network analysis based on synovial tissuesobtained from AIA rats, combining with our previous analysis based on whole blood samples from RA patients with cold-dampness syndrome were performed to identify involved dominant pathways, which were experimentally verified using AIA-wind-cold-dampness stimulation modified (AIA-M) animal model.<b>R</b><b>esults: </b>TZGtreatmentdramatically attenuated joint injury and inflammatory response in AIA rats, and PSMC2-RUNX2-COL1A1 axis, which was closely associated with bone/cartilage damage, was inferred to be one of therapeutic targets of TZG against RA. Experimentally, TZG displayed obvious pharmacological effects for alleviating the joint inflammation and destruction through reinstating the body weight, reducingthe arthritis score, the limbs diameters, the levels of RF and CRP, and the inflammatory cytokines, recovering the thymus and spleen indexes, diminishing bone and cartilage destruction, as well elevating the pain thresholdsof AIA-M rats. In addition, TZG markedly reversed the abnormal energy metabolism in AIA-M rats through enhancing articular temperature, daily water consumption, and regulating expression levels of energy metabolism parameters and hormones. Moreover, TZG also significantly modulated the abnormal expression levels of PSMC2, RUNX2 and COL1A1 proteins in the ankle tissues of AIA-M rats.<b>C</b><b>onclusion: </b>TZG may exert the bone protective effects in RA therapy via regulating bone and cartilage damage-associated PSMC2-RUNX2-COL1A1 axis.

**背景**： 天河追风膏（Tianhe Zhuifeng Gao, TZG）是一款获批的中药专利药物，临床疗效确切，尤其适用于寒湿证类风湿关节炎（Rheumatoid Arthritis, RA）患者，但其潜在药理机制尚未明确。
**方法**： 本研究采用佐剂诱导性关节炎（Adjuvant-induced Arthritis, AIA）大鼠模型评估TZG的抗关节炎活性。通过对AIA大鼠滑膜组织开展转录调控网络分析，并结合本团队此前针对寒湿证RA患者外周血样本的分析结果，筛选出TZG发挥治疗作用的核心通路；随后采用风寒湿刺激修饰的佐剂诱导性关节炎（AIA-wind-cold-dampness stimulation modified, AIA-M）动物模型对上述通路进行实验验证。
**结果**： TZG可显著减轻AIA大鼠的关节损伤与炎症反应，且与骨/软骨损伤密切相关的PSMC2-RUNX2-COL1A1信号轴被推测为TZG治疗RA的潜在靶点之一。实验结果显示，TZG可通过恢复AIA-M大鼠体重、降低关节炎评分与四肢径围、减少类风湿因子（Rheumatoid Factor, RF）和C反应蛋白（C-reactive Protein, CRP）及炎症因子水平、改善胸腺与脾脏指数、减轻骨与软骨破坏，以及提升痛阈，有效缓解关节炎症与组织破坏。此外，TZG可通过升高关节温度、增加每日饮水量、调节能量代谢相关参数与激素的表达水平，显著逆转AIA-M大鼠的异常能量代谢状态。进一步研究表明，TZG可显著调控AIA-M大鼠踝关节组织中PSMC2、RUNX2及COL1A1蛋白的异常表达水平。
**结论**： TZG可通过调控与骨及软骨损伤相关的PSMC2-RUNX2-COL1A1信号轴，在RA治疗中发挥骨保护作用。