伸筋草总生物碱改善大鼠类风湿性关节炎的机制研究

Study on the mechanism of Lycopodium clavatum alkaloids in the improvement of rheumatoid arthritis in rats

目的通过白细胞介素-1受体相关激酶(IRAK1)/肿瘤坏死因子受体相关因子-6(TRAF6)/核转录因子-κB(NF-κB)信号通路探讨伸筋草总生物碱治疗类风湿性关节炎(RA)的机制。方法注射弗氏佐剂建立RA模型。将实验大鼠随机分为正常组、模型组、阳性对照组和伸筋草高、低剂量组,每组10只。给药21 d后,采用容积排水法测定各组大鼠右踝关节肿胀程度;Western Blot法测定大鼠踝关节滑膜组织IRAK1、TRAF6、NF-κB p65蛋白表达;Elisa法测定血清中白细胞介素-1β(IL-1β)和γ干扰素(IFN-γ)的水平。结果与模型组比较,伸筋草高、低剂量组大鼠踝关节肿胀程度明显降低(P<0.05),滑膜组织IRAK1、TRAF6和NF-κB p65蛋白表达显著下调(P<0.05);伸筋草高剂量组大鼠血清中IL-1β和IFN-γ水平显著下降(P<0.05),伸筋草低剂量组IL-1β水平明显降低(P<0.05),IFN-γ水平可见下降趋势。结论伸筋草总生物碱可降低RA大鼠踝关节的肿胀,其作用机制与抑制IRAK1/TRAF6/NF-κB信号通路及其下游炎性因子的分泌密切相关。

Objective To investigate the mechanism of Lycopodium clavatum alkaloids in the treatment of rats with rheumatoid arthritis(RA)via inhibiting interleukin-1(IL-1)receptor associated kinase(IRAK1)/tumor necrosis factor receptor-associated factor-6(TRAF6)/nuclear factor kappa B(NF-κB)signal pathway.Methods Rats were randomly divided into normal group,model group,high and low dose groups of Lycopodium clavatum and positive control group,10 rats in each group.RA model was duplicated by Freund′s complete adjuvant S.C.After 21 d administration,the swelling degree of the right ankle joint was measured by vessel drainage method.Western Blot was used to analyze the expression of IRAK1,TRAF6,NF-κB p65 protein in rat ankle synovium.Also,the level of interleukin-1β(IL-1β)and interferonγ(IFN-γ)in serum was evaluated by Elisa.Results Compared with model group,the swelling degree of ankle joint in Lycopodium clavatum high and low dose group reduced significantly(P<0.05),mean while the expression of IRAK1,TRAF6 and NF-κB p65 protein in synovium decreased markedly(P<0.05).The levels of IL-1βand IFN-γin Lycopodium clavatum high dose group decreased dramatically(P<0.05),while IL-1βin Lycopodium clavatum low dose group decreased significantly(P<0.05).There was a trend of IFN-γto decrease in Lycopodium clavatum low dose group.Conclusion The total alkaloids of Lycopodium clavatum can alleviate the swelling of ankle joint in rats with RA.Its mechanism is closely related to the inhibition of IRAK1/TRAF6/NF-κB signaling pathway and the secretion of inflammatory factors downstreams.