痛风宁对急性痛风性关节炎模型大鼠IL-1β,TNF-α及NALP3炎性体的影响  

Effect of Tongfengning on IL-1β,TNF-α and NALP3 Inflammasome in Model Rats with Acute Gouty Arthritis

作　　者：滕方舟 蔡唐彦 郭洁梅 李保林[1] 王建辉 赖兴泉 张艺强 朱亚菊 黄露露 苏友新 TENG Fang-zhou;CAI Tang-yanI;GUO Jie-mei;LI Bao-lin;WANG Jian-hui;LAI Xing-quan;ZHANG Yi-qiang;ZHU Ya-juI;HUANG Lu-lu;SU You-xin(College of Traditional Chinese Medicine,College of Rehabilitation Medicine,Fufian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fuijiang Health College,Fuzhou 350101,China)

机构地区：[1]福建中医药大学中医学院,康复医学院,福州350122 [2]福建卫生职业技术学院,福州350101

出　　处：《中国实验方剂学杂志》2018年第17期120-125,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81473495)

摘　　要：目的：从核苷酸结合寡聚化结构域样受体蛋白3（NALP3）炎性体角度探讨痛风宁的抗炎作用机制。方法：将48只SD大鼠随机分为造模大鼠36只、正常大鼠12只。造模大鼠采用踝关节局部注射O．2mL尿酸钠（monosodium urate，MSU）混悬液建立急性痛风性关节炎（acute outyarthritis，AGA）模型，正常大鼠予相同部位注射生理盐水。造模成功后，随机分为模型组、痛风宁组、秋水仙碱组，分别用生理盐水、痛风宁药液、秋水仙碱混悬液灌胃，每天2次，共3d。正常大鼠为正常组，同时予等量生理盐水灌胃。造模成功后72h，行腹腔注射麻醉后取踝关节液及滑膜组织。采用酶联免疫吸附测定法（ELISA）检测大鼠踝关节液白细胞介素-1β（IL-lβ），肿瘤坏死因子-a（TNF-a）含量；蛋白免疫印迹法（Westernblot）及实时荧光定量PCR（Real-timePCR）检测滑膜组织NALP3炎性体相关蛋白与mRNA表达。结果：与正常组比较，模型组大鼠踝关节液IL-1β，TNF-α含量显著升高，滑膜组织NALP3，半胱氨酸天冬氨酸蛋白酶-1（Caspase-1），凋亡相关斑点样蛋白（ASC）蛋白及mRNA表达均显著增加（P〈0．01）；与模型组比较，痛风宁组与秋水仙碱组大鼠踝关节液IL-1β，TNF-1β含量显著下降，且滑膜组织中NALP3，Caspase-1，ASC蛋白及mRNA表达均显著降低（P〈0．01）；与秋水仙碱组比较，痛风宁组大鼠踝关节液IL-1β ，TNF-a含量仍较高（P〈0．05），但滑膜组织中NALP3，Caspase-1，ASC蛋白及mRNA表达均较低（P〈0．05，P〈0．01）。结论：痛风宁对AGA的抗炎作用机制可能是通过抑制NALP3炎性体相关蛋白及mRNA表达，从而减少IL-1β，TNF-α的生成，减轻关节炎症反应。Objective： To explore the anti-inflammatory mechanism of Tongfengning from the perspective of nucleotide binding oligomeric domain-like receptor protein 3 （ NALP3 ） inflammasome. Method： A total of 48SD rats were randomly divided into the model rats （n = 36） and the normal rats （n = 12）. The model rats were injected with 0.2 mL monosodium urate （MSU） suspension through the ankle joint to establish the acute gouty arthritis （AGA） model, while the normal rats were injected with saline at the same location. After the AGA model was established successfully, the model rats were randomly divided into model group, Tongfengning group and colchicine group; saline, Tongfengning liquid and colchicine suspension were administered respectively to the three groups by gavage for 3 days, twice a day. The normal rats were taken as normal group, and administered with the equal volume of saline by gavage at the same time. All of the rats received intraperitoneal anesthesia, and their synovial fluid and tissues were collected in 72 h after successful modeling. Enzyme-linked immuno-sorbent assay （ELISA） , Western blot and Real-time PCR technology were used to detect the levels of interleukin-1/3 （IL-113） and tumor necrosis factor-or （TNF-ot） in synovial fluid and the nucleotide binding oligomeric domains-containing protein 3 （NALP3） inflammasome-related protein and mRNA expressions in synovial tissues. Result： Compared with the normal group rats, the levels of IL-Iβ and （TNF-a） in synovial fluid of the model group were increased, and the expressions of NALP3, eystein-aspartic acid protease-1 （ Caspase-1 ） , apoptosis-associated speck-like protein （ASC） and their mRNA in synovial tissues of the model group were increased （P 〈 0.01 ） ; compared with the model group rats, the levels of IL-1/3 and TNF-ot of in synovial fluid of the Tongfengning group and the colchicine group were signifieantly decreased, and the expressions of NALP3, Caspase-1, ASC protein and mRNA in synovial

关 键 词：痛风宁 急性痛风性关节炎 核苷酸结合寡聚化结构域样受体蛋白3(NALP3)炎性体 白细胞介素-1β(IL-1β) 肿瘤坏死因子-α(TNF-α) 

分 类 号：R22[医药卫生—中医基础理论] R24[医药卫生—中医学]