基于Toll样受体/髓系分化初级反应蛋白质88信号通路探讨四妙汤加土茯苓方治疗急性痛风性关节炎的作用机制  

The mechanism of Simiaotang and Tufuling Fang(四妙汤加土茯苓方)for treatment of acute gouty arthritis:a Toll-like receptor/myeloid differentiation primary response protein 88 signaling pathway-based experimental study

作　　者：顾富城 杨美鑫 林惠红 吴伟欣 耿秋东 王和鸣[4] 李楠 GU Fucheng;YANG Meixin;LIN Huihong;WU Weixin;GENG Qiudong;WANG Heming;LI Nan(School of Traditional Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian,China;Zhangzhou Traditional Chinese Medical Hospital,Zhangzhou 363000,Fujian,China;Key Laboratory of Orthopedics&Traumatology and Rehabilitation of Traditional Chinese Medicine of Ministry of Education,Fuzhou 350122,Fujian,China;People’s Hospital Affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian,China)

机构地区：[1]福建中医药大学中医学院,福建福州350122 [2]福建省漳州市中医院,福建漳州363000 [3]中医骨伤及运动康复教育部重点实验室,福建福州350122 [4]福建中医药大学附属人民医院,福建福州350122

出　　处：《中医正骨》2024年第8期9-18,共10页The Journal of Traditional Chinese Orthopedics and Traumatology

基　　金：福建省卫生健康科技计划项目(2021QNA068)。

摘　　要：目的:基于Toll样受体(Toll-like receptor,TLR)/髓系分化初级反应蛋白质88(myeloid differentiation primary response protein 88,MyD88)信号通路探讨四妙汤加土茯苓方治疗急性痛风性关节炎(acute gouty arthritis,AGA)的作用机制。方法:将48只SPF级雄性SD大鼠随机分为6组,每组8只。模型组、秋水仙碱组、中药低剂量组、中药中剂量组、中药高剂量组大鼠以高尿酸造模法结合尿酸盐注射法在右侧踝关节进行AGA造模,空白组以生理盐水灌胃,并在右侧踝关节注射生理盐水。AGA造模结束后,观察大鼠右侧踝关节,评估炎症指数,并计算踝关节肿胀指数。AGA造模结束后24 h,中药低、中、高剂量组大鼠均以四妙散加土茯苓方药液灌胃(生药用量分别为5 g·kg^(-1)、10 g·kg^(-1)、20 g·kg^(-1)),秋水仙碱组以秋水仙碱混悬液灌胃,空白组和模型组大鼠均以等量生理盐水灌胃,每天1次,连续灌胃7 d。药物干预结束后24 h,腹主动脉取血,测定血清尿素氮和肌酐含量;取右侧踝关节滑膜组织,以实时荧光定量聚合酶链反应技术检测TLR/MyD88信号通路相关基因TLR2、TLR4、MyD88、核因子κB抑制剂激酶α(nuclear factor-κB inhibitor kinase-α,IKK-α)、核因子κB抑制剂α(nuclear factor-κB inhibitor-α,IκB-α)mRNA表达水平,以蛋白质印迹技术检测IKK-α、IκB-α蛋白表达水平,采用免疫组化技术观察TLR2、TLR4蛋白表达情况及巨噬细胞聚集情况。结果:(1)模型验证结果。除空白组外,其余5组大鼠右侧踝关节注射尿酸盐混悬液后均逐渐发生肿胀,24 h后炎症指数均达到2级以上。造模后4 h、8 h、12 h、24 h,模型组、秋水仙碱组、中药低剂量组、中药中剂量组、中药高剂量组的踝关节肿胀指数均高于空白组(造模后4 h:P=0.000,P=0.000,P=0.002,P=0.004,P=0.031;造模后8 h:P=0.001,P=0.000,P=0.002,P=0.007,P=0.002;造模后12 h:P=0.000,P=0.000,P=0.004,P=0.010,P=0.006;造模后24 h:P=0.004,P=0.000,P=0.003,P=0.001Objective:To explore the mechanism of Simiaotang and Tufuling Fang(四妙汤加土茯苓方,SMTTFLF)in treatment of acute gouty arthritis(AGA)based on the Toll-like receptor(TLR)/myeloid differentiation primary response protein 88(MyD88)signaling pathway.Methods:Forty-eight specific pathogen-free(SPF)-grade male Sprague-Dawley(SD)rats were selected and randomized into blank group,model group,colchicine group,low-dose SMTTFLF(L-SMTTFLF)group,medium-dose SMTTFLF(M-SMTTFLF)group,and high-dose SMTTFLF(H-SMTTFLF)group,8 cases in each group.All rats but the ones in blank group were intervened by intragastric administration with potassium oxonate(PO)and hypoxanthine(Hx))suspension,followed by intra-articular injection of urate suspension into the right ankle joint for inducing AGA;while the ones in blank group by intragastric administration and intra-articular injection with the same dosage of normal saline.After the end of AGA modeling,the right ankle joint of the rats was observed for assessing the inflammation index,and the ankle swelling index was calculated for confirming whether the AGA models were built successfully.Twenty-four hours after the end of AGA mo-deling,the successfully modeled rats in L-,M-,and H-SMTTFLF group were further intragastric administrated with SMTTFLF in corresponding concentration(consumpting the crude drug as 5,10,and 20 g/kg,respectively),the ones in colchicine group with colchicine suspension,whereas,the ones in blank group and model group with the same dosage of normal saline,once a day for consecutive 7 days.Twenty-four hours after the end of drug intervention,the blood was drawn from the abdominal aorta of rats in each group for detecting the serum le-vels of urea nitrogen and creatinine.After phlebotomizing,the synovial tissues were harvested from the right ankle joints,and the mRNA le-vels of TLR/MyD88 signaling pathway-related genes,including TLR2,TLR4,MyD88,nuclear factor-κB inhibitor kinase-α(IKK-α)and nuclear factor-κB inhibitor-α(IκB-α),were detected by using real-time

关 键 词：关节炎 痛风性 四妙汤 土茯苓 类Toll受体 髓系分化因子88 

分 类 号：R285.5[医药卫生—中药学]