基于VEGF/MAPK/FAK通路调控的风湿骨痛胶囊通络止痛的机制研究

Dredging collaterals and analgesia is a principle treatment for arthralgia-syndrome, how to interpret its scientific connotation of is one of the key issues to be solved. Previous studies have shown that: an often used prescription Fengshi Gutong capsule possess of antarthritic effects for treatment of osteoarthritis in rats by reducing joint injury and cytokines level, chemical constituents were submit to PharmMapper and KEGG bioinformatics softwares for predicting their target proteins and related pathways, the predicted results showed that VEGF, MAPK and focal adhesion (FAK) signaling pathways may be involved in dredging collaterals and analgesia. In this project, reasearch was performed with the guidance of traditional Chinese medicine (TCM) theory and Fengshi Gutong capsule was chosen as the object, rats model of MIA-induced arthritis and rat synovial cell model of TNF-α induced inflammation were made, the material basis and mechanism of dredging collaterals and analgesia base on regulation of angiogenesis and cytokines was investigated using modern advanced technologies (docking, bioactivity based UPLC-Q/TOF screening platform, RT-PCR, ELISA and western blot), with emphasis on VEGF, MAPK and FAK signaling pathways. The project can provide a convincing evidence for scientifically interpreting the essence of dredging collaterals and analgesia and a strategy for illuminating the mechanism of TCM treatment of arthralgia-syndrome.

通络止痛是中医治疗风寒湿痹的重要治则，如何诠释其科学内涵是中医药现代化亟待解决的关键问题之一。本课题组前期研究显示：风湿骨痛胶囊可以有效地抑制骨关节炎大鼠的关节损伤和炎症因子表达；通过Pharm Mapper和KEGG等手段对主要成分的靶点和通路进行分析，提示VEGF、MAPK和focal adhesion(FAK)等信号途径与通络止痛密切相关。本项目在中医理论指导下，以临床上常用的治疗风湿寒痹方剂风湿骨痛胶囊为研究对象，建立大鼠骨关节炎模型及体外滑膜细胞炎症模型，利用反向对接、活性导向的UPLC-Q/TOF筛选平台、ELISA、RT-PCR、western blot等现代先进技术，重点围绕VEGF/MAPK/FAK信号通路，研究风湿骨痛胶囊通过调节血管新生和抑制炎症因子表达治疗风寒湿痹的物质基础及作用机制，为阐释通络止痛的本质提供科学依据，为深入研究中药治疗痹证的分子机理提供思路和方法。

通络止痛是中医治疗风寒湿痹的重要治则，如何诠释其科学内涵是中医药现代化亟待解决的关键问题之一。近来研究表明，炎症因子在骨关节炎疼痛的发展中起着关键作用。本项目以风湿骨痛胶囊为研究对象，从抑制炎症因子的角度出发，旨在阐释中药通络止痛的分子机理。项目研究显示，风湿骨痛胶囊能缓解骨关节炎大鼠膝关节损伤，显著地降低血浆中炎症因子的水平，并能明显抑制IL-1β诱导的滑膜细胞炎症模型中炎症因子的表达；利用IL-1β诱导的人滑膜细胞SW982炎症模型，发现风湿骨痛胶囊中山奈酚、羟基红花黄色素A、乌头碱、齐墩果酸、熊果酸、甘草苷等6个主要抗炎成分；对风湿骨痛胶囊及其主要有效成分进行深入的抗炎机制分析，发现风湿骨痛胶囊通过MAPK，NF-κB，AP-1和Akt等信号通路起到抗炎作用。本项目的研究对风湿骨痛胶囊的质量提升及临床应用有一定的指导意义，为阐释中药通络止痛的科学内涵提供重要的实验依据，为深入解析中药治疗痹证的分子机制提供新的思路和方法。