青藤碱对胶原诱导性关节炎大鼠滑膜血管新生的影响

Effect of Sinomenine on synovial angiogenesis in rats with collagen-induced arthritis

摘要:

目的观察青藤碱对Ⅱ型胶原诱导性关节炎模型大鼠（CIA）低氧诱导因子1α（HIF-1α）、血管内皮生长因子（VEGF）及基质金属蛋白9（MMP-9）的影响，探讨青藤碱对类风湿关节炎（RA）滑膜血管新生的作用机制。方法40 只远交群大鼠，随机选取10 只作为对照组，余30 只大鼠采用尾根部皮下注射Ⅱ型胶原和不完全弗氏佐剂的方法复制关节炎模型。15 d 后造模大鼠随机分为模型组、雷公藤组、青藤碱组，每组10 只。雷公藤组：雷公藤多甙片9.68 mg/（kg·d）灌胃给药；青藤碱组：青藤碱片100 mg/（kg·d）灌胃给药；模型组灌服等体积0.9%氯化钠注射液。1 次/d，共28 d。期间测量大鼠踝关节肿胀度。灌药28 d 后处死大鼠，免疫组织化学法检测关节滑膜组织HIF-1α水平，酶联免疫吸附试验测定血清中VEGF、MMP-9 水平。结果与模型组比较，青藤碱组和雷公藤组大鼠关节肿胀度减轻（p <0.05）；青藤碱组和雷公藤组滑膜组织中HIF-1α 表达降低（p <0.05）；青藤碱组和雷公藤组血清中VEGF、MMP-9 表达降低（p <0.05）。结论青藤碱可降低CIA 踝关节肿胀，疗效与雷公藤多甙相当。其机制可能是通过下调HIF-1α的表达及VEGF、MMP-9水平，从而减少滑膜新生血管生成。

Abstract:

To observe the effect of Sinomenine on the expression of hypoxia-inducible factor-1α(HIF-1α), vascular endothelial growth factor (VEGF) and matrix metalloproteinase 9 (MMP-9) in rats with type Ⅱcollagen-induced arthritis (CIA), and to explore its therapeutic roles and mechanism. Methods Totally 40 male SD rats were recruited, among which 10 were randomly selected as the normal control group. The CIA model was established in the rest 30 rats by subcutaneous injection of type Ⅱcollagen of bovine emulsion in the tail root and induction of incomplete Freund's adjuvant. On day 15 primary immunization rats were randomly divided into 3 groups, i.e. model group, Tripterygium Glycosides (TG) group (at a daily dose of 9.68 mg/kg body weight), and Sinomenine (SIN) group (at a daily dose of 100 mg/kg body weight), 10 in each group. The corresponding medication was given to the rats of each group by gastrogavage once daily for 28 days. An equal volume of sodium chloride injection was given to the rats in the normal control group and the model group by gastrogavage, once daily for 28 successive days. The swelling degree of the joints was measured. The rats were sacrificed after 28 -day treatment. Plasma levels of VEGF and MMP -9 were measured with ELISA. The expression of HIF -1αin synovium was detected by immunohistochemistry. Results Compared with the model group, the swelling degree of the joints was significantly alleviated in the TG group and the SIN group (p < 0.05), the HIF-1αexpression obviously decreased in the synovium of theTG group and the SIN group (p < 0.05), the plasma levels of VEGF and MMP-9 were lower in the TG group and the SIN group (p < 0.05). Conclusions SIN could markedly alleviate the swelling degree of joints in the CIA model rats. Its therapeutic efficacy is equal to that of TG. Its mechanism might be by down-regulating the HIF-1αexpression in joints and the plasma levels of VEGF and MMP-9, thereby reducing synovial neovascularization.