防己黄芪汤通过抑制Notch1通路减轻大鼠胶原诱导性关节炎的机制

Mechanism of Fangji Huangqi Decoction on alleviating collagen-induced arthritis in rats by inhibiting the Notchl pathway

目的:探讨防己黄芪汤(FHD)通过Notch1通路减轻大鼠胶原诱导性关节炎(CIA)大鼠中的机制。方法:采用牛Ⅱ型胶原溶液和完全弗氏佐剂/不完全弗氏佐剂建立CIA模型,FHD低、中、高剂量组分别进行2.2、4.4、8.8 g·kg^(-1)·d^(-1)的FHD干预,每周检测关节炎评分和足关节肿胀度。Masson染色观察关节组织病理形态;ELISA检测大鼠血清白细胞介素(IL)-1β、IL-4、IL-10和IL-17表达;qRT-PCR检测关节和脾脏Notch1、Jagged1、Hes1 mRNA水平,Western Blot检测关节和脾脏Notch1、Jagged1、Hes1和Notch胞内结构域(NICD)蛋白,免疫组化评估关节Notch1、Jagged1、Hes1和NICD的阳性面积。结果:与模型组比较,FHD中、高剂量组大鼠关节炎评分及足关节肿胀度降低(P<0.01),关节破坏减轻,血清促炎细胞因子IL-1β和IL-17降低(P<0.01),抗炎细胞因子IL-4和IL-10升高(P<0.01),关节和脾脏Notch1、Jagged1、Hes1蛋白和mRNA及NICD蛋白表达降低(P<0.01),关节Notch1、Jagged1、Hes1和NICD的阳性面积减少(P<0.01,P<0.05)。结论:FHD通过抑制Notch1信号通路,恢复炎症细胞因子网络平衡,发挥抗关节炎的作用。

Objective:To explore the mechanism of Fangji Huangqi Decoction(FHD)on alleviating collagen-induced arthritis(CIA)in rats through Notchl pathway.Methods:The rat model of collagen-induced arthritis(CIA)was established by injection of type II bovine collagen solution mixed with complete/incomplete Freund's adjuvant.The low,middle and high dose FHD groups were treated with 2.2,4.4,8.8 g·kg^(-1)·d^(-1) FHD respectively.The arthritis score and paw swelling were measured every week.The pathological morphology of joint tissue was observed by Masson staining,the expression of interleukin(IL)-1β,IL-4,IL-10 and IL-17 in serum was detected by ELISA.The mRNA levels of Notchl,Jaggedl and Hesl in joint and spleen were detected by qRT-PCR,Notchl,Jagged1,Hes1 and Notch intracellular domain(NICD)protein in joint and spleen were detected by Western Blot,and the positive areas of Notchl,Jagged,Hesl and NICD in joint were evaluated by immunohistochemistry.Results:Compared with CIA group,in the middle and high dose FHD groups,arthritis scores and foot paw swelling were decreased(P<0.01),joint destruction reduced,IL-1βand IL-17 were decreased(P<0.01),and IL-4 and IL-10 were increased(P<0.01),the protein and mRNA expressions of Notch1,Jaggedl,Hesl,and the protein expression of NICD in the spleen and joints were down-regulation(P<0.01),the positive area of Notch1,Jaggedl,Hesl and NICD were decreased(P<0.01,P<0.05).Conclusion:FHD restore the balance of inflammatory cytokine network and play an anti-arthritis effect by inhibiting Notchl signal pathway.