基于TLR4/NF-κB信号通路探讨通痹胶囊对胶原诱导型关节炎治疗机制的研究

Investigation of therapeutic mechanism of Tongbi capsule in collagen induced arthritis based on TLR4/NF-κB signaling pathway

目的:基于TLR4/NF-κB信号通路研究通痹胶囊对胶原诱导型关节炎(CIA)的治疗机制。方法:48只SD大鼠随机分为对照组、模型组、雷公藤组(0. 01 g/kg)和通痹胶囊低(0. 075 g/kg)、中(0. 150 g/kg)、高(0. 300 g/kg)剂量组,每组8只;使用牛Ⅱ型胶原和完全弗氏佐剂(CFA)建立大鼠CIA模型;采用HE染色观察滑膜组织病理情况,ELISA检测大鼠血清中白介素1β(IL-1β)、白介素6(IL-6)和肿瘤坏死因子α(TNF-α)水平,Western blot检测滑膜组织中Toll样受体4(TLR4)、髓样分化因子88(MyD88)和核因子κB(NF-κB)蛋白表达水平。结果:与正常大鼠相比,模型大鼠关节肿胀度均显著升高,通痹胶囊治疗后关节肿胀度显著降低,模型组滑膜组织增生明显,炎症细胞浸润,关节软骨破坏,通痹胶囊治疗明显改善滑膜组织的增生,降低炎症细胞的浸润;模型组大鼠血清中IL-1β、IL-6、TNF-α水平和滑膜组织中TLR4、MyD88、NF-κB蛋白水平显著升高,通痹胶囊处理后显著降低血清中IL-1β、IL-6、TNF-α水平和滑膜组织中TLR4、MyD88、NF-κB蛋白表达;与通痹胶囊高剂量组相比,TLR4激动剂处理显著提高血清中IL-1β、IL-6、TNF-α水平和滑膜组织中TLR4、MyD88、NF-κB蛋白表达。结论:通痹胶囊可以通过抑制TLR4/NF-κB信号通路,降低炎性反应,显著改善CIA症状。

Objective:To investigate the therapeutic mechanism of Tongbi capsule in collagen-induced arthritis(CIA)based on TLR4/NF-κB signaling pathway.Methods:Forty-eight SD rats were randomly divided into control group,model group,tripterygium wilfordii group(0.01 g/kg)and Tongbi capsule with low dose(0.075 g/kg),middle dose(0.150 g/kg),and high dose(0.300 g/kg)group,8 rats in each group.The CIA model was established by using bovine collagen typeⅡand complete freund’s adjuvant(CFA).HE staining was used to observed the pathological changes of synovial tissue.The serum levels of interleukin 1(IL-1β),IL-6 and Tumor necrosis factorα(TNF-α)were detected by ELISA assay.The protein levels of TLR4,MyD88 and NF-κB in synovial tissue were detected by Western blot.Results:Compared with control group,the arthrocele of rats in model group increased significantly,while the arthrocele of rats decreased significantly after Tongbi capsule treatment.Compared with control group,synovial tissue hyperplasia,inflammatory cells infiltrated,and articular cartilage damage were observed in rats of model group.Tongbi capsule can improve the proliferation of synovial tissue and reduce the infiltration of inflammatory cells.Compared with control group,the serum levels of IL-1β,IL-6,TNF-αand the protein levels of TLR4,MyD88 and NF-κB in synovial tissue were increased significantly in model rats.However,the serum levels of IL-1β,IL-6,TNF-αand the protein levels of TLR4,MyD88 and NF-κB in synovial tissue decreased significantly by Tongbi capsule treatment.Furthermore,compared with Tongbi capsule high dose group,TLR4 agonist treatment significantly increased the serum levels of IL-1β,IL-6,TNF-αand the protein levels of TLR4,MyD 88 and NF-κB in synovial tissue.Conclusion:Tongbi capsule can significantly improve the symptoms of CIA and reduce the inflammatory reaction by inhibiting the TLR4/NF-κB signaling pathway.