基于Toll-MyD88信号通路研究桂枝芍药知母汤治疗痛风性关节炎的作用机制  

Research on Mechanism of Guizhi Shaoyao Zhimu Tang in Treatment of Gouty Arthritis Based on Toll-MyD88 Signaling Pathway

作　　者：王永辉[1] 房树标[1,2] 李艳彦[1] 周然[1] 

机构地区：[1]山西中医学院,太原030024 [2]湖北中医药大学基础医学院,武汉430065

出　　处：《中国实验方剂学杂志》2016年第21期121-126,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：山西省科技创新团队建设项目(2012081018)

摘　　要：目的:观察桂枝芍药知母汤(GD)对尿酸钠致痛风性关节炎(GA)模型大鼠关节滑膜组织中Toll-髓性分化因子88(My D88)信号通路炎性信号表达的影响,探讨其相关的作用机制。方法:180只雄性SD大鼠随机分配到3个实验,分别为关节滑膜免疫组织化学技术(IHC)实验、酶联免疫吸附测定(ELISA)实验、蛋白质免疫印迹(Western blot)实验。各实验取大鼠60只,按体重随机分为6组,每组10只,分别为模型组,正常组,GD高、中、低剂量组(4,8,16 g·kg^(-1)),秋水仙碱阳性药组(3×10^(-4)g·kg^(-1))。实验组均ig给药,正常组、模型组给予等容积的蒸馏水,每天1次,连续给药7 d。第5天ig前,大鼠足踝关节注射尿酸钠悬液诱导GA。取大鼠关节滑膜组织,IHC检测受体Toll样受体-2(Toll-like receptor 2,TLR-2),TLR-4的表达,Image-Pro Plus 6.0图像分析系统测定平均积分吸光度IA,ELISA测定环氧化酶-2(cyclooxygenase-2,COX-2),转化生长因子-β_1(transforming growth factor-β_1,TGF-β_1)表达,Western blot检测My D88,核因子κB酶抑制剂-β(inhibitorκB kinaseβ,IκK-β),核因子κB抑制蛋白α(NF-κB inhibitorα,IκB-α),过氧化物酶体增殖物激活受体-γ(peroxisome proliferator-activated receptor-γ,PPAR-γ)表达水平。结果:造模72 h后,与正常组比较,GA模型大鼠关节滑膜组织中TLR-2,TLR-4平均IA,My D88,IκK-β蛋白及COX-2表达水平明显增高(P<0.05);与模型组比较,GD中、高剂量组TLR-2,TLR-4平均IA,My D88,IκK-β蛋白及COX-2含量表达均明显低于模型组(P<0.05);而TGF-β_1含量及IκB-α,PPAR-γ蛋白表达水平明显增高(P<0.05);GD各剂量组IκK-β蛋白无明显变化。结论:GD治疗GA的作用机制可能与降低TLR-2,TLR-4受体及My D88蛋白表达,增加PPAR-γ,IκB-α表达,抑制NF-κB活化,降低Toll-My D88信号通路炎性因子表达有关。Objective： To investigate the effect of Guizhi Shaoyao Zhimu Tang（ GT） on inflammatory signal expression of Toll-My D88 in synovial tissues of joint of rats with monosodium urate crystal-induced gouty arthritis（ GA）. Method： Totally 180 male SD rats were randomly divided into 3 experiments,namely IHC,ELISA and Western blot experiments,with 60 rats in each experiment. In each experiment,60 rats were included and divided into 6 groups,namely model group,normal group,high,medium and low-dose GD group（ 4,8,16 mg·kg^（-1）） and colchicine group（ 3 × 10^（- 4）g·kg^（-1））,with 10 rats in each group according to weight. All of experimental groups were treated with medicine through gastric administration,normal and model groups were given equal volume of distilled water. Medicine or distilled water was given once daily for consecutively seven days throughout the experiments. On the fifth day,GA model was induced through injection with monosodium urate in the ankle joint cavity of rats. The synovial tissues of the rats were taken,expressions of Toll like receptor-2,4 in synovial tissues of the joint of rats were detected with immunohistochemistry,IA was measured with Image-Pro Plus6. 0 analysis system. ELISA was used to determine expressions of cyclooxygenase-2（ COX-2） and transforming growth factor-β_1（ TGF-β_1）. The expression levels of Myeloid differentiation factor-88（ My D88）,nuclear factor enzyme inhibitor（ IκK-β）,nuclear factor κB inhibitory protein（ IκB-α） and peroxisome proliferator-activated receptor gamma（ PPAR-γ） were detected by Western blot. Result： Compared with normal group after 72 hours,average IA of TLR-2,TLR-4 and the expression level of My D88,IκK-β,COX-2 in synovial tissues of the GA rats significantly increased; compared with normal group,average IA of TLR-2,TLR-4 and the expression level of My D88,IκK-β,COX-2 significantly decreased in medium and high-dose GT groups（ P〈0. 05）,whereas TGF-β_1,IκB-α and PPAR-γ protein expre

关 键 词：桂枝芍药知母汤 痛风性关节炎 Toll-髓性分化因子88 关节滑膜组织 

分 类 号：R285.5[医药卫生—中药学]