芍药苷对胶原诱导性关节炎大鼠滑膜细胞Ras和Raf-1表达的影响

Effect of paeoniflorin on the expressions of Ras and Raf-1 in the synoviocytes of rats with collagen-induced arthritis

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.