N-9,10-蒽醌-2-甲酰基苯丙氨酸治疗大鼠痛风性关节炎的作用及机制  

Effect and mechanism of N-9,10-anthraquinone-2-formyl phenylalanine on gouty arthritis in rats

作　　者：王伟[1] 高天舒 汤同军 贾昱晗 裴乃琪 钱有桥 赵嘉鑫 丁骁鹏 葛昊 张波 李洪雷 王星 WANG Wei;GAO Tianshu;TANG Tongjun;JIA Yuhan;PEI Naiqi;QIAN Youqiao;ZHAO Jiaxin;DING Xiaopeng;GE Hao;ZHANG Bo;LI Honglei;WANG Xing(Department of Orthopedics,The People′s Hospital of Jurong,Jurong 212400,China;Department of Pharmacology,School of Pharmacy,China Pharmaceutical University,Nanjing 211198,China;School of Basic Medicine and Clinical Pharmacy,China Pharmaceutical University,Nanjing 211198,China;School of Traditional Chinese Pharmacy,China Pharmaceutical University,Nanjing 211198,China;Department of Pharmacy,Kangda College of Nanjing Medical University,Lianyungang 222000,China)

机构地区：[1]句容市人民医院骨科,江苏句容212400 [2]中国药科大学药学院药理系,江苏南京211198 [3]中国药科大学基础医学与临床药学学院,江苏南京211198 [4]中国药科大学中药学院,江苏南京211198 [5]南京医科大学康达学院药学部,江苏连云港222000

出　　处：《药学研究》2023年第5期303-309,共7页Journal of Pharmaceutical Research

基　　金：2018年度镇江市社会发展指导性项目(No.FZ2018014);2018年度句容市民生科技计划项目(No.SF2018013987);江苏省基础研究计划(自然科学基金)-青年基金项目(No.BK20180574);南京医科大学康达学院2021年度科研发展基金(No.KD2021KYJJZD009)。

摘　　要：目的探究N-9,10-蒽醌-2-甲酰基苯丙氨酸(NAY)对尿酸钠(MSU)所致痛风性关节炎(GA)大鼠的作用及机制。方法将雄性SD大鼠随机分为7组,每组8只,即正常组、模型组、NAY低(20 mg·kg^(-1))、中(40 mg·kg^(-1))、高剂量组(80 mg·kg^(-1))、秋水仙碱组(0.8 mg·kg^(-1))和别嘌呤醇组(10 mg·kg^(-1)),通过关节腔内注射MSU晶体建立大鼠急性GA模型,各组灌胃给药5 d,给药第3天造模。将原代培养的大鼠腹部巨噬细胞分成空白组、模型组、NAY组、MCC950组和联合给药组。软尺测量大鼠踝关节周长并计算大鼠关节肿胀度;检测大鼠血清尿酸(UA)、肌酐(CRE)和尿素氮(BUN)含量;HE染色观察大鼠踝关节滑膜组织病理切片;检测各组血清、关节滑膜和细胞上清液中肿瘤坏死因子-α(TNF-α)和白介素-1β(IL-1β)的含量;并检测各组关节滑膜和细胞中NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)和半胱氨酸天冬氨酸蛋白酶-1(caspase-1)的蛋白表达。结果动物实验发现,与模型组相比,NAY给药组在造模12、24和48 h均可不同程度改善GA大鼠踝关节肿胀度,且NAY各个剂量组均显著降低GA大鼠血清UA、CRE和BUN水平,同时显著降低GA大鼠血清中TNF-α和IL-1β水平及GA大鼠关节滑膜中炎症因子TNF-α和IL-1β水平,而NAY能显著降低GA大鼠关节滑膜中NLRP3、ASC和caspase-1的蛋白表达,HE染色结果显示NAY可减轻GA模型大鼠踝关节滑膜组织炎性细胞浸润,改善细胞形态。细胞实验发现,NAY、MCC950和联合给药组均可显著提高MSU诱导大鼠巨噬细胞活力,降低上清液TNF-α和IL-1β的含量,并显著降低模型组NLRP3、ASC和caspase-1蛋白表达,但是联合给药组与MCC950组相比,上述指标没有显著性差异。结论NAY可在体内和体外有效预防MSU晶体诱导的GA,机制可能与其降尿酸和抑制NLRP3炎症小体表达有关。Objective To explore the effect and mechanism of N-9,10-anthraquinone-2-formyl phenylalanine(NAY)on gouty arthritis(GA)rats induced by sodium urate(monosodium urate).Methods Male SD rats were randomly divided into 7 groups:8 each,namely normal group,model group,NAY low(20 mg·kg^(-1)),medium(40 mg·kg^(-1)),high dose group(80 mg·kg^(-1)),colchicine group(0.8 mg·kg^(-1))and allopurinol group(10 mg·kg^(-1)).Rat acute GA model was established by intra-articular injection of MSU crystals.Each group was administered for 5 days and molded on the third day.The primary cultured rat abdominal macrophages were divided into blank group,model group,NAY group,MCC950 group,co-administration group.Measure the circumference of the rat ankle joint with a soft ruler and calculate the swelling degree of the rat joint.Serum samples of rats were collected to detect the contents of serum uric acid(UA),creatinine(CRE)and urea nitrogen(BUN).HE staining was used to observe the pathological sections of the synovium of the ankle joint of rats.The contents of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)were measured in serum,synovial membrane and cell supernatant of each group.The protein expressions of NOD-like receptor family pyrin domain-containing 3(NLRP3),apoptosis-associated speck-like protein containing CARD(ASC)and cysteinyl aspartate-specific proteases-1(caspase-1)were detected in synovium and cells of each group.Results As found in animal experiments,the NAY administration group could improve the ankle swelling of GA rats in different degrees at 12,24 and 48 hours,and each dose group of NAY can significantly reduce serum UA,CRE and BUN levels in GA rats,while significantly reducing TNF-αand IL-1βlevels in serum with the decrease of TNF-αand IL-1βlevels in the synovial membrane.In addition,NAY could significantly reduce the protein expression of NLRP3,ASC and Caspase-1 in the synovium of the ankle joint of GA rats.HE staining results showed that NAY can reduce inflammatory cell infiltration and improve cell mo

关 键 词：痛风性关节炎 N-9 10-蒽醌-2-甲酰基苯丙氨酸 黄嘌呤氧化酶抑制剂 尿酸钠 NLRP3炎症小体 

分 类 号：R965[医药卫生—药理学]