痹通对佐剂性关节炎大鼠TLR4/MYD88/NF-κB信号通路的影响

Effect of Bitong Decoction(痹通)on TLR4/MYD88/NF-κB Signaling Pathway in Adjuvant Arthritis Rats

目的探讨中药痹通对佐剂性关节炎(AA)大鼠TLR4/MYD88/NF-κB信号通路的影响。方法制备AA大鼠模型,反转录聚合酶链式反应法(RT-PCR)检测足跖部Toll样受体4(TLR4)、髓样分化因子88(MYD88)的表达;酶联免疫吸附法(ELISA)检测外周血核因子κB(NF-κBp65)、白介素10(IL-10)的水平;Western Blot法检测足跖软组织中肿瘤坏死因子α(TNF-α)表达,免疫组化法(IHC)检测踝关节环氧合酶2(COX-2)表达。结果与正常组比较,模型组TLR4、MYD88、NF-κBp65、TNF-α、COX-2表达均显著增高(P<0.05);IL-10显著降低(P<0.05)。与模型组比较,痹通高剂量组、中剂量组TLR4、MYD88、NF-κBp65、TNF-α、COX-2表达均显著降低(P<0.05),IL-10表达显著提高(P<0.05)。结论痹通能有效降低AA大鼠TLR4、MYD88、NF-κBp65、TNF-α、COX-2等炎症蛋白的表达,提高IL-10的表达,其治疗作用机制可能与降低TLR4/MYD88/NF-κB信号通路的表达有关。

Objective To explore the effect of Bitong Decoction(痹通)on TLR4/MYD88/NF-κB signaling pathway in adjuvant arthritis(AA)rats.Methods The AA rat model was prepared.The expression of Toll-like receptor 4(TLR4)and myeloid differentiation factor 88(MYD88)were detected by reverse transcription polymerase chain reaction(RT-PCR).Enzyme-linked immunosorbent assay(ELISA)method was used to detect the levels of nuclear factor-κB(NF-κB p65)and interleukin 10(IL-10)in peripheral blood.Western Blot(WB)method to detect the expression of tumor necrosis factor-α(TNF-α)in the soft tissues of the plantar.Immunohistochemistry method(IHC)to detect COX-2 expression in ankle joints.Results Compared with the normal control group,the expressions of TLR4,MYD88,NF-κB p65,TNF-α,and COX-2 in the model control group were significantly increased(P<0.05);IL-10 was significantly decreased(P<0.05).Compared with the model control group,the expressions of TLR4,MYD88,NF-κB p65,TNF-αand COX-2 in the 41.0 g/kg and 20.4 g/kg groups of Bitong Decoction were significantly reduced(P<0.05),and the expression of IL-10 was significantly increased(P<0.05).Conclusions Bitong Decoction can effectively reduce inflammatory proteins expression of TLR4,MYD88,NF-κBp65,TNF-α,COX-2 in AA rats,and increase the expression of IL-10.Its therapeutic mechanism may be related to the reduction of the TLR4/MYD88/NF-κB signaling pathway expression.