蛋白酶激活受体1在甲型流感病毒感染所致炎症中的作用机制  

The role and mechanism of PAR1 in the inflammation caused by IAV infection

作　　者：高蕊[1] 杨亚荣[1] 袁娟[1] 潘建丽[2] GAO Rui;YANG Ya-rong;YUAN Juan;PAN Jian-li(Outpatient of Infectious Diseases,Xi'an Children's Hospital,Xi'an 710003,Shaanxi,China;Special Department,Xi'an Children's Hospital,Xi'an 710003,Shaanxi,China)

机构地区：[1]西安市儿童医院感染门诊,陕西西安710003 [2]西安市儿童医院特需病区,陕西西安710003

出　　处：《川北医学院学报》2023年第8期1026-1029,共4页Journal of North Sichuan Medical College

基　　金：陕西省自然科学基础研究计划(S2022-JC-QN-1494)。

摘　　要：目的:探究蛋白酶激活受体1(PAR1)在甲型流感病毒(IAV)感染所致的炎症中的作用机制。方法:选取10例甲型流感病毒患者及10名健康受试者作为研究对象,比较二者血清PAR1表达水平。按照不同感染复数将人肺癌细胞系A549细胞分为control组、0.05组、0.1组、0.2组及0.4组,使用MTT法检测各组细胞活性;使用酶联免疫吸附试验(ELISA)检测各组白细胞介素6(IL-6)水平;使用RT-qPCR检测各组PAR1 mRNA水平;Western blot检测PAR1、Toll样受体3(TLR3)及核因子κB(NF-κB) p65蛋白水平。敲低PAR1或过表达TLR3后,使用Western blot检测各组细胞PAR1、TLR3、NF-κB p65、p-NF-κB p65蛋白表达量。结果:PAR1在IAV感染患者血清及IAV感染的A549细胞中表达上调。与control组细胞相比,IAV感染细胞中细胞活性降低,促炎因子IL-6水平升高,且PAR1的mRNA表达水平上调。si-PAR1抑制了IAV诱导的炎症细胞因子IL-6水平的升高。TLR3过表达逆转了由si-PAR1所致的TLR3、p-NF-κB p65蛋白水平降低(P<0.05)。TLR3过表达逆转了si-PAR1所致的细胞活力和炎性因子的水平(P<0.05)。si-PAR1通过介导TLR3/NF-κB信号通路降低IAV感染所致的炎症反应。结论:敲低PAR1可通过抑制TLR3/NF-κB信号通路降低了IAV感染所致的炎症反应,发挥抗病毒作用。Objective:To explore the role and mechanism of protease activated receptor 1(PAR1)in inflammation caused by influenza A virus(IAV)infection.Methods:10 patients with IAV and 10 healthy subjects were selected as the research objects,and the serum PAR1 expression levels were compared between them.Human lung cancer cell line A549 cells were divided into control group,0.05 group,0.1 group,0.2 group,and 0.4 group according to different infection multiples.Cell activity in each group was detected using MTT method.The interleukin-6(IL-6)levels in each group was detected by enzyme-linked immunosorbent assay(ELISA),RT-qPCR was used to detect the mRNA expression level of PAR1.Western blot was used to detect the expression level of PAR1,TLR3,and NF-κB p65.After knocking down PAR1 or overexpressing TLR3,Western blot was used to detect PAR1,TLR3,and NF-κB p65,p-NF-κB p65 protein expression level in each groups.Results:PAR1 was up-regulated in the serum of patients infected and A549 cells infected with IAV.Compared with the cells of the control group,IAV infected cells showed decreased cell activity,increased levels of pro-inflammatory factor IL-6,and up-regulated mRNA expression levels of PAR1.si-PAR1 inhibited the elevation of inflammatory cytokine IL-6 levels induced by IAV.Overexpression of TLR3 reversed the decrease of protein level of TLR3,p-NF-κB p65 caused by si-PAR1(P<0.05),overexpression of TLR3 also reversed the cell viability and inflammatory factor levels caused by si-PAR1(P<0.05).si-PAR1 mediated the TLR3/NF-κB signaling pathway to reduce the inflammatory response caused by IAV infection.Conclusion:Knockdown of PAR1 reduced the inflammatory response caused by IAV infection via inhibiting the TLR3/NF-κB signaling pathway and exerted an antiviral effect.

关 键 词：甲型流感病毒 蛋白酶激活受体1 TLR3/NF-κB 炎症 

分 类 号：R373.1[医药卫生—病原生物学] R511[医药卫生—基础医学] R363.1