豨莶草对大鼠佐剂型关节炎的治疗作用及机制研究

Therapeutic Effect and Underlying Mechanism of Herba Siegesbeckiae in the Treatment of Adjuvant Arthritis in Rats

目的:通过佐剂型大鼠关节炎模型探讨豨莶草对类风湿关节炎的治疗作用及机制研究。方法:豨莶草药材粉碎后乙醇加热回流制成干浸膏。60只雄性大鼠随机分为6组:正常对照组、模型组、美洛昔康组(40.0 mg·kg^(-1))、豨莶草高、中、低剂量组(1.6,0.8,0.4 g·kg^(-1)),每组10只。通过注射完全弗氏佐剂进行刺激,构建佐剂型关节炎模型,造模成功后美洛昔康和豨莶草组灌胃给药,模型组和正常对照组给予等体积生理盐水,1次/d,持续28 d。测量大鼠足跖肿胀度、脾脏脏器系数,HE染色观察关节滑膜组织病理变化,Western blot检测关节滑膜NLRP3炎性小体的表达,ELISA检测血浆白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)、干扰素-γ(IFN-γ)、白细胞介素18(IL-18)的表达。结果:与正常对照组比较,模型组大鼠足跖肿胀度和脾脏系数明显增大,关节滑膜内NLRP3蛋白表达及血浆IL-1β、IL-6、TNF-α、CRP、IFN-γ、IL-18明显升高(P<0.05)。与模型组比较,豨莶草各剂量组大鼠足跖肿胀度和脾脏系数明显减少,关节滑膜内NLRP3蛋白表达及血浆IL-1β、IL-6、TNF-α、CRP、IFN-γ、IL-18明显降低(P<0.05)。结论:豨莶草高、中剂量组均可能通过抑制NLRP3炎性小体的表达,从而抑制滑膜细胞增殖,调节炎性因子生成,进而抑制类风湿关节炎的炎性反应,以达到减轻关节损伤的作用。

Objective: To investigate the therapeutic effect and mechanism of Herba siegesbeckiae in rheumatoid arthritis by adjuvant arthritis model in rats. Methods: Herba siegesbeckiae was crushed and then the dry extract was prepared by heating reflux with ethanol. Sixty rats were randomly divided into six groups: normal control group,model group,meloxicam group(40.0 mg·kg^(-1)),Herba siegesbeckiae high,medium and low dose groups(1.6,0.8 and 0.4 g·kg^(-1)) with 10 ones per group. Adjuvant arthritis models were established by injecting Freund’s complete adjuvant. After the successful modelling,meloxicam and Herba siegesbeckie groups were with intragastric administration,and the model group and normal control group were given equal volume of normal saline,once daily for 28 days. The voixpedis swelling and spleen coefficient in rats were measured,the pathological changes of synovium were observed by HE staining,the expression of NLRP3 inflammasome in synovium was detected by Western blot,and the expressions of IL-1β,IL-6,TNF-α,CRP,IFN-γ and IL-18 in plasma were detected by ELISA. Results: Compared with that in the normal control group,the paw swelling degree and the spleen coefficient of rats were significantly increased,and the expression of NLRP3 protein in the synovium of joints and plasma IL-1β,IL-6,TNF-α,CRP,IFN-γ and IL-18 were significantly increased in the model group(P<0.05). Compared with those in the model group,the paw swelling degree and the spleen coefficient of rats were significantly decreased,and the expression of NLRP3 protein in the synovium of joints and plasma IL-1β,IL-6,TNF-α,CRP,IFN-γ and IL-18 were significantly decreased in all Herba siegesbeckie groups(P<0.05). Conclusion: The Herba siegesbeckiae high and medium dose groups can inhibit the expression of NLRP3 inflammasome to inhibit the proliferation of synovial cells and regulate the production of inflammatory cytokines,and then inhibit the inflammatory response of rheumatoid arthritis resulting in the reduction of joint injury.