“祛风湿、止痹痛、补肝肾、益气血”治法是中医骨科治疗膝痹的经典治法，其代表方独活寄生汤治疗KOA的临床疗效确切，特色优势明显，但其作用机制不清。最新研究表明Zinc-ZIP8-MTF1轴在KOA发病过程中起重要作用，课题组前期研究发现独活寄生汤对软骨细胞ZIP8蛋白具有抑制作用。据此，提出研究假说，认为独活寄生汤可通过影响ZIP8、ZNT9及Mt1/Mt2的表达，维持软骨细胞Zn2+稳态，抑制MTF1和基质金属蛋白酶的转录与激活，即整体调控Zinc-ZIP8-MTF1轴的活化，达到延缓关节软骨退变，改善骨关节炎症状和病情的作用。本研究将模拟炎性细胞因子环境，针对独活寄生汤成分复杂，作用靶点多样等特点，采用基因沉默、基因过表达等技术排除多靶点效应干扰，旨在探寻独活寄生汤的作用靶点和作用规律，探索并验证其对关节软骨细胞Zinc-ZIP8-MTF1轴调控的作用机制。

The therapeutic method of dispelling wind and dampness, relieving the impediment and arthralgia, nourishing the Liver and Kidney, replenishing qi and blood and its representative decoction Duhuojisheng Decoction are the classical therapy and decoction in Chinese medicine orthopaedic respectively, whose clinical effects is curative, feature and advantage is obvious in treating KOA,however, the mechanism of its action and its biological foundation are still unknown.The preliminary study of the research group shows the Zinc-ZIP8-MTF1 axis plays an important role in the pathogenesis of KOA, which locates the upstream of MMPs,and Zn2+ is the important structural composition and active center of MMPs,so the abnormal regulating of Zinc-ZIP8-MTF1 axis can lead to activating of MMPs.So our task group considered that the abnormal metabolic status of Zn2+ in the knee joint cartilage is one of the key factors of secondary injury of articular cartilage,Duhuojisheng decoction can regulate the expression of ZIP8, ZNT9 and MTF1,inhibition of MMPs activation.This project plan to use fluorescence quantitative qPCR Technology and gene knockout technology in vitro experiments. Objective evaluation the effect of Duhuojisheng Decoction on Zn2+ metabolism of knee joint cartilage ,to explore the new targets of Duhuojisheng Decoction in the treatment of knee osteoarthritis .