补肾活血汤干预瘦素调控骨关节炎软骨、滑膜及脂肪细胞交互作用的网络机制研究

Osteoarthritis (OA), as the most common form of arthritis, is a disease with formidable impact on human healthy. Traditional Chinese medicine treatment for OA has long history, and has distinct characteristics and advantages. We have identified that Bushen Huoxue decoction had significant clinical efficiency in OA therapy. It has been verified by experimental studies in rat and rabbit model that Bushen Huoxue decoction delayed joint cartilage degeneration by improving micro-circulation and regulating the intra-articular micro-environment, while the molecular mechanism was unclear. Recent studies indicated that leptin may be one of the central adipokine mediators being investigated for metabolic-osteoarthritis link. In addition to classic pro-inflammatory factors, such as IL-1 and TGF-β, Leptin over-expressed in local joint of OA patients specifically, and contributed to OA pathobiology. Leptin is unique among other adipokines analyzed to date in that only leptin has been found to exist at higher concentrations in the synovial fluid compared to serum. These facts pointed out that Leptin may be new biomarkers or molecular targets in OA diagnosis and therapy. It has been reported that Bushen Huoxue decoction could effectively reduce serum leptin level in polycystic ovary syndrome (PCOS) rat model, while the function of this Chinese herbal involved in articular micro-environment regulation was unknown. In this study, we would continue to identify the expression pattern of a set of OA-associated cytokines, proteases, and adipokines before and after Bushen Huoxue decoction treatment to assess the molecular mechanism of Bushen Huoxue decoction in OA therapy by using proteomic, genomic and systems biology approaches, and analyze the Bushen Huoxue decoction-inducd cellular responses and the cross-talk networks of OA articular cells, including cartilage, synovial and lipocytes. The results of this study will construct the OA-associated cytokins, proteases and adipokines data pool, provide theories for clinical application of Bushen Huoxue decoction, and will be helpful for our understanding of the onset and the progression of OA.

骨关节炎（OA）是严重危害人类健康的慢性骨关节疾病，中药治疗OA有悠久历史和独特优势，并已取得良好临床疗效。我课题组在前期研究中证实中药方剂补肾活血汤能有效改善OA临床症状，具显著疗效。依据相关动物实验文献，补肾活血汤能改善OA局部微循环，调节理化环境，延缓软骨退变，但作用机制尚待揭示。新近研究发现除IL-1、TGF-β等经典炎症相关因子外，脂肪因子瘦素特异性地以显著高于血循环的浓度在OA局部高表达，参与OA病理过程，有可能成为OA治疗靶分子。在多囊卵巢实验大鼠模型中已证实补肾活血方能调控机体脂肪代谢并下调瘦素表达；但其对骨关节微环境的调控作用未见研究。本申请拟以软骨、滑膜及脂肪细胞交互作用网络为研究对象，通过组学、系统生物学大规模数据分析及分子生物学技术研究补肾活血汤干预OA的分子机制。课题完成不仅能为临床用药提供科学依据，还将推进对OA发生发展机制的认识，促进临床辩证论治。

骨关节炎（OA）是严重危害人类健康的慢性骨关节疾病。本课题以脂肪因子中重要分子瘦素（Leptin）为切入点，通过蛋白组学、系统生物信息学分析，结合分子生物学技术研究探索了软骨、滑膜及脂肪细胞交互作用网络在OA患者中的表达，搜索以瘦素为代表的脂肪因子在OA中的差异表达基因及信号转导途径，探讨了他们在OA发病中的作用，并在此基础上开展了补肾活血汤靶向瘦素干预OA的分子机制及作用研究，为OA的病因病机研究和疾病的治疗提供了新的实验数据和理论指导。课题组的研究内容及重要成果如下：①课题以脂肪因子Leptin为切入点，研究鉴定了Leptin调控的细胞因子信号网络并探讨其在骨关节炎发生发展中的作用。依照项目书的任务要求，课题组研究发现了不同OA阶段软骨的瘦素、脂联素、抵抗素表达水平存在差异。不同损伤程度软骨的瘦素的表达水平可互相影响，血浆瘦素、脂联素表达水平的增高与关节液中瘦素、脂联素表达水平的升高有关。瘦素、脂联素可能会独立地对OA软骨产生影响。血浆中的瘦素、脂联素浓度分别与关节液中的瘦素、脂联素的浓度密切相关。②在上述研究基础上，我们鉴定了OA不同程度病损软骨中差异表达的蛋白分子，并筛选可能与Leptin相关、参与细胞间通讯及信号转导的信号通路。其中最令我们感兴趣的是Wnt/β-catenin信号通路。我们现已在细胞模型中发现证实Leptin能够调控Wnt/β-catenin信号通路，这一调控作用可能是通过PI3K/Akt及JAK-STAT3两条信号通路完成的。这一发现揭示了OA发生发展的新途径，为临床OA诊治提供新的思路和理论指导，具有潜在的临床价值。③补肾活血汤及其有效成分补骨脂素对OA具有治疗作用。课题完成过程中，我们除了对前期工作中研究的Leptin及Wnt/β-catenin信号通路间cross-talk进行了进一步验证外，还通过细胞模型及动物模型探索了补肾活血汤及补骨脂素对OA的增殖与信号转导作用，结果发现补骨脂素促进体外软骨细胞模型的增殖；补肾活血汤通过Wnt/β-catenin信号通路调控金属基质蛋白酶的表达，这一结果为补肾活血方在OA治疗中的临床应用及新药的研发提供了新的信息和研究思路。