NLRP3炎症小体抑制剂N14对小鼠痛风性关节炎的治疗作用  

Therapeutic effects of the NLRP3 inflammasome inhibitor N14 in the treatment of gouty arthritis in mice

作　　者：姜晓琳 郭凯 贺玉伟 陈怡铭 杜姗姗 江余祺 李卓悦 李长贵[3] 秦冲 JIANG Xiao-lin;GUO Kai;HE Yu-wei;CHEN Yi-ming;DU Shan-shan;JIANG Yu-qi;LI Zhuo-yue;LI Chang-gui;QIN Chong(School of Chemical Engineering,Qingdao University of Science and Technology,Qingdao 266042,China;Key Laboratory of Marine Drugs,Chinese Ministry of Education,School of Medicine and Pharmacy,Ocean University of China,Qingdao 266003,China;Shandong Provincial Clinical Research Center for Immune Diseases and Gout,the Affiliated Hospital of Qingdao University,Qingdao 266555,China;Center for Targeted Protein Degradation and Drug Discovery,Ocean University of China,Qingdao 266003,China;Laboratory for Marine Drugs and Bioproducts,Qingdao National Laboratory for Marine Science and Technology,Qingdao 266003,China;Marine Biomedical Research Institute of Qingdao,Qingdao 266071,China)

机构地区：[1]青岛科技大学化工学院,山东青岛266042 [2]中国海洋大学医药学院,海洋药物教育部重点实验室,山东青岛266003 [3]山东省免疫性疾病与痛风临床医学研究中心,青岛大学附属医院,山东青岛266555 [4]中国海洋大学蛋白质靶向降解与药物研发中心,山东青岛266003 [5]青岛海洋科学技术国家实验室海洋药物与生物制品实验室,山东青岛266003 [6]青岛海洋生物医药研究院,山东青岛266071

出　　处：《药学学报》2024年第5期1229-1237,共9页Acta Pharmaceutica Sinica

基　　金：山东省自然科学基金优秀青年基金项目(ZR2021YQ53)。

摘　　要：探讨Nod样受体蛋白3(Nod-like receptor protein 3,NLRP3)炎症小体抑制剂N14对尿酸钠(mono sodium urate,MSU)晶体诱导的痛风性关节炎(gouty arthritis,GA)小鼠的治疗作用。首先采用细胞计数试剂(cell counting kit-8,CCK-8)法检测N14对小鼠单核巨噬细胞J774A.1活力的影响;利用免疫印迹法(Western blot)检测N14对细胞上清中成熟的白介素1β(interleukin 1β,IL-1β)、胱天蛋白酶-1(cysteinyl aspartate specific proteinase-1,caspase-1)活化物caspase-1 p20及细胞裂解液中NLRP3、pro-caspase-1和pro-IL-1β表达的影响。通过MSU诱导的小鼠痛风性关节炎模型,检测痛风性关节炎小鼠的红、肿、热、痛反应;苏木素-伊红(hematoxylin-eosin,H&E)染色进行小鼠足部病理学检测;应用免疫印迹法检测小鼠足部组织中NLRP3、pro-caspase-1和pro-IL-1β的表达;并考察N14对小鼠血浆中谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate transaminase,AST)等含量的影响。本文中所有动物实验均获得青岛海洋生物医药研究院科学伦理委员会批准(批准号:E-MBWNL-2024-20)。实验结果显示,J774A.1细胞与高浓度的N14(100μmol·L-1)共孵育后未见明显细胞毒性,且有效阻止MSU诱导的NLRP3炎症小体的激活。在小鼠痛风性关节炎模型中,N14显著改善痛风性关节炎引起的红、肿、热、痛,下调小鼠足部组织中NLRP3炎症小体相关蛋白水平,同时对小鼠血浆中各项生化指标无显著影响,具有良好的耐受性。综上,N14通过抑制NLRP3炎症小体通路有效缓解小鼠痛风性关节炎,对相关疾病的预防和治疗都具有重要意义。Monosodium urate(MSU)-induced the gouty arthritis(GA)model was used to investigate the effect of Nod-like receptor protein 3(NLRP3)inhibitor N14 in alleviating GA.Firstly,the effect of NLRP3inhibitor N14 on the viability of mouse monocyte macrophage J774A.1 was examined by the cell counting kit-8(CCK-8)assay.The expression of mature interleukin 1β(IL-1β)and cysteinyl aspartate specific proteinase-1(caspase-1)p20 in the cell supernatant and the expression of NLRP3,caspase-1 and pro-IL-1βproteins in the cell lysates was detected by Western blot for the inhibitory effect of N14 on the MSU-induced NLRP3 inflammasome activation in J774A.1 cells.Animal behavioral tests were used to detect redness,swelling,heat and pain in mice with gouty arthritis.Hematoxylin-eosin(H&E)staining revealed pathologic changes and inflammatory infiltration in foot sections.Protein expression of NLRP3,caspase-1,and pro-IL-1βin mouse hind paw tissues were assessed by Western blot.The effect of N14 on the plasma levels of alanine transaminase(ALT),aspartate transaminase(AST),creatinine(CRE),urea,and uric acid(UA)was investigated by the MSU-induced gouty arthritis model in mice.All animal experiments in this paper were approved by the Scientific Ethics Review Board of Qingdao Marine Biomedical Research Institute(grant No.E-MBWNL-2024-20).The experimental results showed that N14did not exhibit cytotoxicity in mouse monocyte macrophage J774A.1 cells at concentrations up to 100μmol·L-1,and N14 effectively prevented MSU-induced activation of NLRP3 inflammasome.In the mouse gouty arthritis model,N14 significantly ameliorated the redness,swelling,heat and pain caused by GA,and down-regulated the levels of NLRP3 inflammasome-associated proteins in mouse hind paw tissues.Meanwhile,N14 appeared to be well tolerated,as it did not significantly affect various biochemical indices in mouse plasma.In conclusion,N14effectively alleviated GA in mice by inhibiting the NLRP3 inflammasome pathway,which is important for both prevention and treatment of relat

关 键 词：4-((二甲基氨基)甲基)-N-((1 2 3 5 6 7-六氢-s-茚-4-基)氨基甲酰基)苯磺酰胺 痛风性关节炎 尿酸钠结晶 炎症因子 NOD样受体蛋白3炎症小体 白细胞介素-1Β 

分 类 号：R967[医药卫生—药理学]