摘要
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 protein α(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 cattle Ⅱ collagen 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,IκB α,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.
引文
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