雷公藤内酯醇对类风湿关节炎大鼠TLR4/NF-κB信号通路的调控作用研究  

Regulation of tripterygiumwilfordii lactone alcohol on TLR4/NF-κB signaling pathway in rheumatoid arthritis rats

作　　者：陈颖婷[1] 何柯新 王云秀[1] 何敏[1] 郑智明[1] CHEN Yingting;HE Kexin;WANG Yunxiu;HE Min;ZHENG Zhiming(Department of Clinical Laboratory,Guangdong Province Traditional ChineseMedical Hospital,Guangzhou,Guangdong 510120,China;Department of Clinical Laboratory,Guangzhou Huiai Hospital,Guangzhou,Guangdong 510108,China)

机构地区：[1]广东省中医院检验科,广东广州510120 [2]广州市惠爱医院检验科,广东广州510108

出　　处：《国际检验医学杂志》2019年第17期2053-2057,共5页International Journal of Laboratory Medicine

基　　金：广东省中医药局科研项目(20161082);广东省科技计划项目(2014A020212502)

摘　　要：目的 观察雷公藤内酯醇治疗类风湿关节炎(RA)大鼠的疗效,并通过Toll-样受体4(TLR4)/核因子κB(NF-κB)信号通路探讨其作用机制。方法 将24只健康SD大鼠,随机分为空白对照组、RA模型组、阳性药双氯芬酸组、雷公藤内酯醇组,每组6只。采用热杀死结核分枝杆菌诱导RA模型组、阳性药双氯芬酸组、雷公藤内酯醇组大鼠制作RA模型;造模成功后,阳性药双氯芬酸组给予10mg/kg双氯芬酸灌胃,雷公藤内酯醇组大鼠给予6mg/kg雷公藤内酯醇灌胃,空白对照组和RA模型组大鼠给予等量0.9%氯化钠溶液灌胃。采用ELISA法测定大鼠血清中炎性细胞因子[肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-4、IL-6]表达水平;评价关节炎指数及足体积。免疫组织化学法检测关节滑膜组织TLR4、NF-κB、p-NF-κB蛋白表达,q-PCR法检测滑膜组织TLR4mRNA的表达。结果 与空白对照组比较,RA模型组血清中TNF-α、IL-4、IL-6表达水平显著升高(P<0.05);与RA模型组比较,阳性药双氯芬酸组血清中TNF-α、IL-4、IL-6表达水平显著下降(P<0.05),且雷公藤内酯醇组TNF-α、IL-4、IL-6表达水平呈极显著下降(P<0.01)。与RA模型组比较,雷公藤内酯醇组在第8、12、16、20天时关节炎指数和足体积显著降低(P<0.05),阳性药双氯芬酸组在第12、16、20天时关节炎指数和足体积显著降低(P<0.05)。与空白对照组比较,RA模型组滑膜组织中TLR4、NF-κB、p-NF-κB蛋白表达量和TLR4mRNA表达量显著上调(P<0.05);与RA模型组比较,阳性药双氯芬酸组滑膜组织中TLR4、NF-κB蛋白表达量和TLR4mRNA表达量显著下降(P<0.05),雷公藤内酯醇组滑膜组织中TLR4、NF-κB、p-NF-κB蛋白表达量和TLR4mRNA表达量显著下调(P<0.05)。结论 雷公藤内酯醇可能通过抑制TLR4/NF-κB信号通路的信号因子表达,从而降低炎性因子的产生,减少炎性反应,起到治疗RA的作用。Objective To observe the therapeutic effect of tripterygiumwilfordii lactone alcohol on rheumatoid arthritis (RA) rats,and explore its mechanism through Toll-like receptor 4 (TLR4)/nuclear factor κB (NF-κB) signaling pathway. Methods Twenty-four healthy SD rats were randomly divided into blank control group,RA model group,positive drug diclofenac group and tripterygiumwilfordii lactone alcohol group with 6 rats in each group.Rats in RA model group,positive drug diclofenac group and tripterygiumwilfordii lactone alcohol group were induced by heat killing Mycobacterium tuberculosis to make RA model.After successful modeling,positive drug diclofenac group was given 10 mg/kg diclofenac acid by gavage,and tripterygiumwilfordii lactone alcohol group was given 6 mg/kg tripterygiumwilfordii lactone alcohol by gavage,rats in blank control group and RA model group were given the same amount of 0.9% sodium chloride solution by gavage.The levels of inflammatory cytokines (TNF-α,IL-4 and IL-6) in serum of rats were determined by ELISA,and the arthritis index and foot volume were evaluated.The expression of TLR4,NF-κB and p-NF-κB in synovial tissue was detected by immunohistochemistry and the expression of TLR4 in synovial tissue was detected by q-PCR. Results Compared with the blank control group,the serum levels of TNF-α,IL-4 and IL-6 in RA model group were significantly decreased ( P <0.05).Compared with RA model group,the expression levels of TNF-α,IL-4 and IL-6 in serum of diclofenac group decreased significantly ( P <0.05),and the expression levels of TNF-α,IL-4 and IL-6 in tripterygiumwilfordii lactone alcohol group decreased significantly ( P <0.01).Compared with RA model group,the arthritis index and foot volume of tripterygiumwilfordii lactone alcohol group decreased significantly on the 8th,12th,16th and 20th day ( P <0.05),while the arthritis index and foot volume of positive drug diclofenac group decreased significantly on the 12th,16th and 20th day ( P <0.05).Compared with the blank control group,the expr

关 键 词：雷公藤内酯醇 内风湿性关节炎 Toll-样受体4/核因子κB信号通路 

分 类 号：R593.22[医药卫生—内科学]