目的:研究芍药苷(paeoniflorin,Pae)对胶原诱导性关节炎(collagen—induced arthritis,CIA)大鼠滑膜细胞Ras和Raf-1表达的影响。方法:Ⅱ型胶原诱导大鼠CIA模型,Pae(25:50,100mg·kg^-1)灌胃给药,qd,连续7d,测量足肿胀及体重变化,进行关节炎评分。分离滑膜细胞与不同浓度Pae共培养,Western blot检测Ras和Raf-1的水平。结果:CIA大鼠致敏后d16出现足爪肿胀,d20~d28肿胀达峰值。关节炎指数增高,并伴有体重下降。Pae(25,50,100mg·kg^-1)可改善足爪炎症,降低关节炎指数,且Pae50和100mg·kg^-1组大鼠体重恢复正常。CIA大鼠滑膜细胞中Ras和Raf-1的表达水平高于正常大鼠,致敏后d20~d28表达水平最高,d42后开始下降。整体用药Pae(50,100mg·kg^-1)可降低Ras表达;Pae(25,50,100mg·kg^-1)能降低Rail表达水平。体外用药Pae(2.5,12.5,62.5mg·L^-1)可降低CIA大鼠滑膜细胞中Ras和Rail的表达水平。结论:Ras和Raf-1介导的信号通路参与CIA大鼠滑膜炎症,Pae通过影响Ras和Raf-1的表达而调节异常的Ras—MAPKs信号通路可能是其抑制滑膜炎症的主要机制之一。
Objective: To investigate the effect of paeoniflorin (Pae) on the expressions of Ras and Raf-1 in the synoviocytes of the rats with collagen-induced arthritis (CIA). Methods: After CIA was induced by type Ⅱ collagen, rats were treated with oral Pae (25, 50 and 100 mg·kg^-1). Synoviocytes were isolated and cultured with Pae at various concentrations. Ras and Raf-1 were detected by Western blot. Results: CIA induced paw swelling, increased arthritis index (AI) and decreased body weight. Pae diminished the paw swelling and AI at 25, 50 and 100 mg·kg^-1 , and increased the weight of CIA rats at 50 and 100 mg·kg^-1. The expressions of Ras and Raf-1 in synoviocytes were increased in CIA rats, which reached the maximum at 20 - 28 d and declined from 42 d after induction of CIA. Pae at 50, 100 mg·kg^-1 suppressed the expressions of Ras and Raf-1 in the synoviocytes in vivo. In cultured synoviocytes, Pae at 2.5, 12.5 and 62.5 mg·L^-1 also inhibited the expressions in vitro. Conclusion: The signals, Ras and Raf-1, are involved in synovitis of CIA rats. Pae can modulate the abnormal Ras-MAPK signal pathway via affecting the expression of Ras and Raf-1, which may be one of the mechanisms underlying its inhibition of synovitis.