温阳通络方抗胶原诱导性关节炎小鼠滑膜新生血管形成的机制研究

Study on the Mechanism of Wenyang Tongluo Recipe against Synovial Neovascularization in Collagen induced Arthritis Mice

目的探讨温阳通络方对抗胶原诱导性关节炎(collagen-induced arthritis,CIA)小鼠滑膜新生血管形成的作用机制。方法复制胶原诱导型关节炎小鼠,随机分为甲氨蝶呤组、模型组、空白对照组、温阳通络方高、中、低剂量组,每组5只。测量足趾容积,采用HE染色、免疫组化、Western blot检测滑膜组织中Ang-2、VEGF蛋白表达情况。结果造模前(0 d)、造模35 d,各组DBA/1小鼠足趾容积差异无统计学意义;造模48 d,各组足趾容积与空白对照组比较差异有统计学意义(P<0.01)。空白组、甲氨蝶呤组、温阳通络方高、中、低剂量组与模型组比较,差异有统计学意义(P<0.05)。空白组、模型组、温阳通络方低、中剂量组与甲氨蝶呤组比较,差异有统计学意义(P<0.01),温阳通络高剂量组与甲氨蝶呤组比较,差异无统计学意义。模型组滑膜组织HE染色提示有大量淋巴细胞浸润及滑膜细胞增生;甲氨蝶呤组、温阳通络方高、中、低剂量组仅有少量淋巴细胞,未见明显滑膜细胞增生。模型组小鼠滑膜组织中有大量Ang-2、VEGF蛋白表达,而甲氨蝶呤组、温阳通络方高、中、低剂量组中Ang-2、VEGF蛋白表达明显减少(P<0.01)。Western blot检测,空白组、甲氨蝶呤组、温阳通络方高、中、低剂量组Ang-2、VEGF蛋白表达水平均较模型组明显降低。结论温阳通络方能抗胶原诱导性关节炎小鼠滑膜组织新生血管形成,进而减轻滑膜炎症。

Objective To investigate the mechanism of Wenyang Tongluo Recipe against synovial neovascularization in mice with collagen-induced arthritis(CIA).Methods The mice with CIA were randomly divided into methotrexate group,model group,blank control group,Wenyang Tongluo Recipe of high,middle and low dose groups,with 5 mice in each group.Measuring toe volume,HE staining and the expressions of Ang-2 and VEGF in synovium were detected by immunohistochemistry and Western blot.Results There were no significant differences in toe volume of DBA/1 mice in each group before treatment(0 d)and on 35 d,while significant differences in toe volume when compared with the blank control group on 48 d(P<0.01).Compared with the model group,the blank group,methotrexate group,Wenyang Tongluo Recipe of high,middle and low dose groups had significant differences(P<0.01).There was no difference in methotrexate group and high dose group.HE staining of synovium tissues in model group showed a lot of lymphocyte infiltration and synovium proliferation,while methotrexate group,Wenyang Tongluo Recipe of high,middle and low dose groups showed only a small amount of lymphocyte,and no obvious synovium proliferation.There were a lot of Ang-2 and VEGF protein expressions in model group,and the expression levels of Ang-2 and VEGF decreased in methotrexate group,Wenyang Tongluo Recipe of high,middle and low dose groups(P<0.01).Western blot showed that the levels of Ang-2 and VEGF in blank group,methotrexate group,Wenyang Tongluo Recipe of high,middle and low dose groups were significantly lower than those in model group.Conclusion Wenyang Tongluo Recipe could prevent the formation of new blood vessels in synovium of collagen-induced arthritis mice,thus reduce synovitis.