从ERK通路探讨复方雷公藤外敷对类风湿关节炎滑膜血管形成的影响

At present it still lacks of radical cure methods in rheumatoid arthritis (RA) on clinical and the target for its therapy is controlling inflammation, relieving bone erosion as possible，and this target need to be achieved by the inhibition of synovial cell proliferation, pannus angiogenesis. In the pathogenesis of RA, extracellular signal-regulated kinase (ERK) can be activated by cytokines, the activation of ERK pathway and IL-1, TNF- α, vascular endothelial growth factor (VEGF) and angiopoietin (Ang) participate in RA synovial proliferation, angiogenesis, and finally resulting in bone erosion. Tripterygium wilfordii is an effective traditional Chinese medicine in the treatment of RA, however it can cause severe toxic side effects by oral administration. So the Tripterygium wilfordii external treatment is becoming new clinical focus, while previous studies lacked of its mechanism research.This study discuss the effect of external application of Tripterygium wilfordii on the expression of IL-1, TNF-α, VEGF and Ang in synovial tissue and serum of CIA rats, as well as the expression of ERK on fibroblast-like synoviocytes (FLS), and the relationship of between ERK pathway activation and synovial proliferation as well as angiogenesis, to explore the mechanism of external application of Tripterygium wilfordii, to enrich the connotation of TCM theory, to provide theoretical basis for its clinical application, to provide new ideas and methods for the study of traditional Chinese medicine.

目前临床仍缺乏根治类风湿关节炎（RA）的有效方法，以尽可能控制炎症、减轻骨侵蚀为治疗目标，这需通过抑制滑膜细胞增殖、血管翳形成而实现。在RA发病中细胞外信号调节激酶（ERK）可由细胞因子激活，ERK通路活化与IL-1、TNF-α、血管内皮生长因子（VEGF）和血管生成素（Ang）等共同参与RA滑膜增殖、血管生成，最终导致骨侵蚀。雷公藤是治疗RA的有效中药，但口服可致严重毒副作用，故雷公藤外治正成为临床新的关注点，而以往缺少关于其外治作用机制研究。本研究拟从复方雷公藤外敷对CIA大鼠滑膜组织和血清中IL-1、TNF-α、VEGF和Ang表达，以及对成纤维样滑膜细胞（FLS）中ERK的影响方面，通过研究ERK通路活化与滑膜增殖、血管生成的关系，探讨其外治的作用机制，以诠释RA中医外治法疗效机制特点，丰富中医基础理论的内涵，为其临床应用提供理论依据，为中医药研究提供新的思路和方法。

本课题通过复方雷公藤外敷剂对CIA模型大鼠滑膜组织ERK通路活化与IL-1β、TNF-α、VEGF和Ang-1的关系，探讨复方雷公藤外敷RA的抗炎、镇痛与减轻血管翳形成作用的信号转导机制。采用II型胶原诱导型关节炎（CIA）大鼠模型，治疗组用扶他林外敷、雷公藤多苷片口服和复方雷公藤外敷。同正常组、CIA模型组比较，观察复方雷公藤外敷组CIA大鼠关节肿胀度，关节、肝、肾、卵巢病理切片，血清、滑膜组织及成纤维样滑膜细胞的白介素1β（IL-1β）、肿瘤坏死因子α（TNF-α）、血管内皮生长因子（VEGF）和血管生成素1（Ang-1）含量差异以及滑膜组织与成纤维样滑膜细胞的细胞外信号调节激酶（ERK）通路的变化情况。结果表明：复方雷公藤外敷剂通过降低CIA大鼠血清及滑膜组织炎症细胞因子水平减弱ERK通路活化，进而抑制滑膜组织增殖，减少滑膜中血管新生和血管翳形成，且无肝、肾及生殖毒性。本研究结果诠释了复方雷公藤外敷剂治疗RA的抗炎与抑制滑膜增殖，减少滑膜中血管新生作用的内在机制，丰富中医基础理论的内涵，为其临床应用提供理论依据，为中医药研究提供新的思路和方法。