小柴胡汤对CFA大鼠滑膜组织NF-κB信号通路作用的探讨

Effect of Xiao Chaihutang on NF-κB Signaling Pathway in Synovial Tissue of CFA Rats

目的:观察小柴胡汤治疗后弗氏完全佐剂(CFA)大鼠滑膜组织核转录因子(NF)-κB信号通路中肿瘤坏死因子受体相关死亡结构域(TARDD),NF-κB抑制蛋白α(IκBα),IκB激酶-α(IKKα),NF-κB p65蛋白的表达。方法:将SPF级健康成年雌性Wistar大鼠应用弗氏完全佐剂和牛Ⅱ型胶原制备CFA动物模型。根据随机数字表,将大鼠随机分为正常组、模型组、小柴胡汤低、中、高剂量组、雷公藤多苷组。各组于造模后7 d开始灌胃给药,正常组及模型组均予注射用水,小柴胡汤低、中、高剂量组(5. 94,11. 88,23. 76 g·kg-1),雷公藤多苷片(0. 006 3 g·kg-1),分别2 mL/次,每天3次灌胃,连续给药28 d后观察实验大鼠踝关节组织病理学及蛋白免疫印迹法(Western blot)检测NF-κB信号通路中TARDD,IKKα,IκBα,NF-κB p65蛋白表达。结果:与正常组比较,模型组大鼠右后踝关节组织病理评分显著增加(P <0. 01),NF-κB信号通路中TARDD,IKKα,IκBα,NF-κB p65蛋白表达显著增高(P <0. 01)。与模型组比较,随着小柴胡汤剂量的增加,实验大鼠右后踝关节组织病理评分显著减少(P <0. 01),在大鼠踝关节组织标本NF-κB信号通路中TARDD,IKKα,IκBα,NF-κB p65关键蛋白的表达上,小柴胡汤低剂量组蛋白表达明显减低(P <0. 05),而小柴胡汤中剂量组、小柴胡汤高剂量组、雷公藤多苷组蛋白表达显著减低(P <0. 01)。结论:研究显示小柴胡汤随着剂量的增加,可以有效地改善滑膜炎症,抑制NF-κB炎症信号通路中关键蛋白的表达,从而阻止炎症而抑制骨侵蚀。

Objective:To observe the expression of tumor necrosis factor receptor-associated death domain(TARDD),nuclear transcription factor-κB inhibiting proteinα(IκBα)IκB kinase-α(IKKα)and nuclear transcription factor(NF)-κB p65 protein in the NF-κB signaling pathway of synovial tissues of complete Freund’s adjuvant(CFA)rats after treatment with Xiao Chaihutang(XCHT).Method:In animal experiments,SPF health adult female Wistar rats were used to prepare the CFA animal model of rats with rheumatoid arthritis with Freund’s complete adjuvant and cattleⅡcollagen type.According to the random number table,the rats were randomly divided into the normal group,the model group,the low-dose XCHT group,the medium-dose XCHT group,the high-dose XCHT group,and the Tripterygium glucosides group.The drugs were given at 7 d after the model was built.Both normal group and model group were given water for injection,and low-dose XCHT group(5.94 g·kg-1),medium-dose XCHT group(11.88 g·kg-1),high-dose XCHT group(23.76 g·kg-1),Tripterygium glucosides group(0.006 3 g·kg-1)were given corresponding drugs by gavage for three times a day,2 m L/time.The histopathology of rat ankle joint was observed,and the protein expressions of TARDD,IKKα,IκBα,NF-κB p65 in the NF-κB signaling pathway in synovial tissue of CFA rats were detected by Western blot.Result:With the increase of the dosage of XCHT,the histopathological score of the right posterior ankle joint of the experimental rats was increased.And in the protein expressions of TARDD,IKKα,IκBα,NF-κB p65 in NF-κB signaling pathway in Synovial Tissue of CFA rats,compared with the model group,the statistical results of the lowdose XCHT group showed decreased protein expressions(P<0.05)and significant differences.However,the statistical results in the medium-dose XCHT group,the high-dose XCHT group and the tripterygium glucosides group showed decreased protein expression(P<0.01)and significant differences.Compared with the normal group,the histopathological score of the right posterior ankle of the model group was significantly increased(P<0.01),and the protein expressions of TARDD,IKKα,IκBα,NF-κB p65 in the NF-κB signaling pathway were significantly increased(P<0.01).Compared with model group,with the increase of dose of Xiao Chaihutang,histopathologic score of posterior ankle of experimental rats significantly reduced(P<0.01).In rats ankle tissue samples,TARDD,IKKα,IκBα,NF-κB p65 key protein expressions in the NF-κB signaling pathway and protein expressions in low-dose XCHT group were obviously lower(P<0.05),and protein expressions in the medium-dose XCHT group,the high-dose XCHT group and the Tripterygium glucosides group were significantly lower(P<0.01).Conclusion:This study shows that as the dose of Xiao Chaihutang increases,it could effectively improve synovitis,and suppress the expressions of key proteins in the inflammatory signaling pathway of NF-κB,thereby preventing inflammation and suppressing bone erosion.