清热除痹方对大鼠痛风性关节炎NALP3炎性体信号通路的影响  

Effect of Qingre Chubi Decoction on NALP3 Inflammasome Signaling Pathway in Rats with Gouty Arthritis

作　　者：石尉宏 劳贝妮 张娴娴[1] 韩冷[1] 曾富玲 曹诗丹 徐伟[1] 孙维峰[1] SHI Weihong1,2, LAO Beini3, ZHANG Xianxian1, HAN Lengl, ZENG Fuling1,2, CAO Shidan2, XU Weil, SUN Weifeng1(1. Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010 Guangdong, China; 2. Guangzhou University of Chinese Medicine, Guangzhou 510006 Guangdong, China; 3. Guangdong Second Hospital of Traditional Chinese Medicine, Guangzhou 510095 Guangdon)

机构地区：[1]广州军区广州总医院,广东广州510010 [2]广州中医药大学,广东广州510006 [3]广东省第二中医院,广东广州510095

出　　处：《中药新药与临床药理》2018年第4期461-467,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：军队中医药研究重大专项(10ZYZ116);广州市科技项目(201607010122)

摘　　要：目的探讨清热除痹方治疗大鼠痛风性关节炎(GA)以及对NALP3炎性体信号通路的影响。方法将162只大鼠随机分为6组,每组27只,分别为正常组,模型组,清热除痹方高、中、低剂量组(22,11,5.5g·kg^(-1)·d^(-1)),秋水仙碱阳性药物组(0.2 mg·kg^(-1)·d^(-1))。实验组分别用药物灌胃,模型组和正常组均给予等体积蒸馏水灌胃,连续灌胃11 d,于第8 d除正常组外所有大鼠均采用尿酸钠膝关节腔注射法进行模型复制,观察模型复制后24 h大鼠步态分级。并于模型复制后24,48,72 h各时间点每组取9只大鼠心脏采血3 m L并处死大鼠,ELISA法测定血清中肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)含量。并于相应时间点取大鼠膝关节滑膜组织。Western Blot检测滑膜Nod样受体蛋白3(NALP3)、炎性体凋亡相关斑点样蛋白(ASC)和半胱氨酸天冬氨酸酶-1(caspase-1)的蛋白表达,ELISA测定滑膜IL-1β含量。模型复制后72 h,采用苏木素-伊红(H&E)染色观察大鼠膝关节滑膜组织病理学改变。结果与正常组比较,模型组大鼠步态显著变差(P<0.01),滑膜组织炎症反应明显,各时间点模型组血清中炎性因子TNF-α,IL-1β含量显著升高(P<0.01),滑膜组织中NALP3、ASC、caspase-1、IL-1β表达明显升高(P<0.01);与模型组比较,清热除痹方高、中、低剂量组和秋水仙碱组步态均得到明显改善(P<0.01),滑膜组织炎症反应不同程度减轻,各时间点血清中TNF-α、IL-1β含量显著降低(P<0.05,P<0.01);各时间点滑膜NALP3、ASC、caspase-1、L-1β表达明显降低(P<0.05,P<0.01)。结论清热除痹方能够通过抑制NALP3炎性体信号通路有效治疗GA大鼠。Objective To investigate the application of Qingre Chubi decoction on treatment of gouty arthritis（GA）in rats and its effects on NALP3 inflammasome signaling pathway. Methods A total of 162 healthy mice were randomly divided into six groups with 27 rats in each group,including normal group,model group,Qingre Chubi decoction high dose group（22 g·kg^（-1）·d^（-1））,medium dose group（11 g·kg^（-1）·d^（-1））,low dose group（5.5 g·kg^（-1）·d^（-1））,and colchicine group（0.2 mg·kg^（-1）·d^（-1））. Medicine or distilled water was given once daily for eleven consecutive days throughout the experiment. On the eighth day,GA model was made by injection of monosodium urate in the knee joint cavities of rats. At the 24 ^（th）,48 ^（th）,72 ^（th） hours after modeling,9 rats were executed by drawing out 3 m L blood from their hearts,and the synovial tissue of the knee joint of rats were taken. Levels of tumor necrosis factor-α（TNF-α） and interleukin-1β（IL-1β） in serum were determined by enzyme-linked immunosorbent assay（ELISA）. Rat synovial tissues were obtained at corresponding time points. The synovial tissues of the rats were taken and protein expression of Nod-like receptor protein 3（NALP3）, apoptosis-associated speck-like protein（ASC） and caspase-1 were detected with Western Blot. Expression of inflammatory cytokine IL-1β in synovial tissues was detected with ELISA. At 72 th hour after model establishment,hematoxylin-eosin（HE） staining was used to observe the histopathological changes in synovial tissues of rats＇ knees. Results As compared with the normal group, the gait of rats in the model group was significantly worse（P〈0.01）, and the inflammatory response of the synovial tissue was obvious. The levels of inflammatory cytokines TNF-α and IL-1β in the model group were significantly increased at sampling time points（P〈0.01）. The expression levels of NALP3 in the synovial tissue including NALP3,ASC,caspase-1 and L-1β were

关 键 词：清热除痹方 NALP3 炎性体 痛风性关节炎 

分 类 号：R285.5[医药卫生—中药学]