消退素D1对H1N1流感病毒FM1感染小鼠肺组织的保护作用及机制  

Protective Effect and Mechanism of Action of Resolvin D1 on the Lung Tissue of Mice Infected with H1N1 Influenza Virus FM1

作　　者：丁凌[1] 冷报浪[1] 董洪英[1] 王海燕[1] DING Ling;LENG Baolang;DONG Hongying;WANG Haiyan(Hangzhou Third People’s Hospital,Hangzhou 310009,China)

机构地区：[1]杭州市第三人民医院,杭州310009

出　　处：《病毒学报》2023年第4期962-970,共9页Chinese Journal of Virology

基　　金：杭州市科技计划引导项目(项目号:20211231Y040),题目:Notchl介导的肺血管内皮细胞凋亡干预慢性阻塞性肺疾病的作用机制研究。

摘　　要：NOD样受体热蛋白结构域相关蛋白3(NOD⁃like receptor pyrin domain⁃containing 3,NLRP3)炎症小体的激活在H1N1流感病毒感染引起肺组织损伤中起关键作用;消退素D1(Resolvin⁃D1,RvD1)是具有抗炎活性的脂质介质,在多种脏器损伤模型中通过抑制NLRP3炎症小体的激活减轻组织损伤及炎症反应。为了研究RvD1对H1N1感染小鼠肺组织的保护作用及机制,本研究以雄性C57BL/6小鼠为对象、通过H1N1/FM1病毒株滴鼻的方式进行H1N1感染的建模,给予RvD1腹腔注射、空白(Negative control,NC)慢病毒或NLRP3慢病毒尾静脉注射干预。检测肺指数,肺组织病理改变,肺组织中病毒载量及NLRP3、凋亡相关斑点样蛋白(Apoptosis⁃associated speck⁃like protein,ASC)、Cleaved caspase⁃1的表达水平,支气管肺泡灌洗液(Bronchoalveolar lavage fluid,BALF)中IL⁃1β、IL⁃18的含量。与对照组比较,模型组小鼠肺组织出现了肺泡结构损伤、炎症细胞浸润等病理改变,肺指数及肺组织中病毒载量、NLRP3、ASC、Cleaved caspase⁃1的表达水平、BALF中IL⁃1β、IL⁃18的含量增加(P<0.05);与模型组比较,RvD1组小鼠肺组织的病理改变减轻,肺指数及肺组织中NLRP3、ASC、Cleaved caspase⁃1的表达水平、BALF中IL⁃1β、IL⁃18的含量降低(P<0.05),病毒载量无明显变化(P>0.05);与NC慢病毒+RvD1组比较,NLRP3慢病毒+RvD1组小鼠肺组织病理改变加重,肺指数及肺组织中NLRP3、ASC、Cleaved caspase⁃1的表达水平、BALF中IL⁃1β、IL⁃18的含量增加(P<0.05)。以上结果表明RvD1通过抑制NLRP3炎症小体减轻H1N1/FM1感染小鼠的肺损伤。Activation of the NLR family pyrin domain containing 3(NLRP3)inflammasome has a key role in lung⁃tissue injury caused by infection by the H1N1 influenza virus.Resolvin D1(RvD1)is a lipid mediator with anti⁃inflammatory activity.In various models of organ injury,RvD1 has been shown to reduce tissue injury and the inflammatory response by inhibiting activation of the NLRP3 inflammasome.We wished to study the protective effect and mechanism of action of RvD1 in the lung tissue of H1N1⁃infected mice.Male C57BL/6 mice were used to establish a model of H1N1 infection by nasal drip of the H1N1/FM1 virus strain.Then,NC lentivirus or NLRP3 lentivirus was injected(i.v.)into the caudal vein.The Lung Index,pathological changes in lung tissue,viral load,as well as expression of NLRP3,apoptosis⁃associated speck⁃like protein(ASC),and cleaved caspase⁃1 in lung tissue were measured.Also,the contents of interleukin(IL)⁃1βand IL⁃18 in bronchoalveolar lavage fluid(BALF)were measured.Compared with the control group,lung tissue showed pathological changes(alveolar⁃structure injury and infiltration of inflammatory cells),the Lung Index,viral load,expression of NLRP3,ASC,cleaved caspase⁃1 in lung tissue,and the contents of IL⁃1βand IL⁃18 in BALF increased in the model group(P<0.05).Compared with the model group,the pathological changes of lung tissue improved,the Lung Index,expression of NLRP3,ASC,and cleaved caspase⁃1 in lung tissue,and the contents of IL⁃1βand IL⁃18 in BALF decreased(P<0.05),but the viral load did not change significantly(P>0.05)in the model group.Compared with the NC lentivirus+RvD1 group,the pathological changes in lung tissue were aggravated,expression of NLRP3,ASC,and cleaved caspase⁃1 in lung tissue and contents of IL⁃1βand IL⁃18 in BALF increased in the NLRP3 lentivirus+RvD1 group(P<0.05).Overall,our results suggested that RvD1 reduces lung injury in H1N1/FM⁃infected mice by inhibiting expression of the NLRP3 inflammasome.

关 键 词：甲型流感病毒 H1N1/FM1病毒株 消退素D1 NLRP3炎症小体 肺损伤 炎症反应 

分 类 号：R373.1[医药卫生—病原生物学]