五灵胶囊通过调控TLR4/NF-κB通路防治急性痛风性关节炎作用的研究

Wuling Capsules prevent acute gouty arthritis by inhibiting TLR4/NF-κB pathway

目的 探讨五灵胶囊对由单尿酸钠晶体(monosodium urate,MSU)诱导的急性痛风性关节炎(acute gouty arthritis,AGA)大鼠的治疗作用。方法 通过踝关节腔内注射MSU构建AGA大鼠模型。用缚线法、足趾容积测量仪、双足平衡测痛仪检测踝关节周径、足趾容积、双足痛觉;用HE染色观察大鼠右侧踝关节滑膜病理变化;用免疫吸附试验(enzyme-linked immunesorbent assay,ELISA)检测血清中炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素1β(interleukin-1β,IL-1β)水平;用实时荧光PCR技术分别检测关节滑膜中的炎症因子及TLR4/NF-κB信号通路关键指标的表达情况。结果 与对照组比较,模型组的踝关节周径、足趾容积、对痛觉的敏感程度均显著升高(P<0.05),滑膜组织炎性浸润明显,血清中炎症因子TNF-α、IL-1β的水平均显著升高(P<0.05),滑膜组织中炎症因子及TLR4/NF-κB信号通路中关键指标的表达量均显著升高(P<0.05);与模型组比较,秋水仙碱和五灵胶囊各剂量组均可明显改善由MSU诱导的AGA大鼠踝关节周径、足趾容积及对痛觉的敏感程度(P<0.05),改善滑膜组织炎性浸润,抑制由MSU诱导的AGA大鼠血清中炎症因子的上调(P<0.05),抑制关节滑膜组织中IL-1β、TNF-α、IL-6及TLR4、MyD88、NF-κB、IκBα、NLRP3的表达(P<0.05)。结论 五灵胶囊可能通过减少炎症因子的释放,抑制TLR4/NF-κB信号通路,发挥防治AGA的作用。

Objective To investigate the effect of Wuling Capsules on acute gouty arthritis(AGA)induced by monosodium urate(MSU)in rats.Methods AGA rat model was constructed by injecting MSU into the joint cavity.The ankle joint circumference,toe volume and foot pain perception were measured by string,toe volume and foot balance measuring instrument.The pathological changes of the synovium of the right ankle joint was observed by HE staining.The serum levels of inflammatory cytokines,tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)were detected by enzyme-linked immunesorbent assay(ELISA).The expressions of inflammatory factors and key indicators of TLR4/NF-κB signaling pathway in joint synovium were detected by realtime fluorescence poly chain reaction(RT-PCR).Results Compared with blank group,the ankle circumference,toe volume and sensitivity to pain in model group were significantly increased(P<0.05),the inflammatory infiltration of synovial showed remarkable lesions,and the serum levels of inflammatory factors TNF-αand IL-1βwere significantly increased(P<0.05).The expressions of inflammatory factors and key indexes of TLR4/NF-κB signaling pathway in synovial tissue were significantly up-regulated(P<0.05).Compared with model group,colchicine and Wuling Capsules could significantly improve the ankle circumference,toe volume and sensitivity to pain of AGA rats(P<0.05),improve inflammatory infiltration of synovial tissue,and inhibit the up-regulation of inflammatory factors in serum of AGA rats(P<0.05).The expression of IL-1β,TNF-α,IL-6,TLR4,MyD88,NF-κB,IκBαand NLRP3 in synovial tissue was inhibited(P<0.05).Conclusion Wuling Capsules might attenuate AGA mainly through reducing the release of inflammatory factors by inhibiting TLR4/NF-κB signaling pathway.