甲型流感病毒感染老年小鼠支气管肺泡灌洗液中细胞因子变化与精氨酸代谢的相关性研究  

The correlation between cytokines and arginine metabolism in bronchoalveolar lavage fluid of aged mice infected with the influenza A virus

作　　者：周娟娟 刘诗洋 李雪 杨欣欣 杨俊连 罗声栋 陈威巍 许文 王福生 Zhou Juanjuan;Liu Shiyang;Li Xue;Yang Xinxin;Yang Junlian;Luo Shengdong;Chen Weiwei;Xu Wen;Wang Fusheng(Graduate School of Bengbu Medical University,Bengbu 233030,China;Department of Infectious Diseases,the Fifth Medical Center,Chinese People′s Liberation Army(PLA)General Hospital,Beijing 100039,China;School of Pharmaceutical Sciences,Binzhou Medical College,Yantai 264003,China)

机构地区：[1]蚌埠医科大学研究生院,蚌埠233030 [2]中国人民解放军总医院第五医学中心感染病医学部研究所,北京100039 [3]滨州医学院药学院,烟台264003

出　　处：《中华实验和临床病毒学杂志》2024年第4期422-431,共10页Chinese Journal of Experimental and Clinical Virology

基　　金：国家重点研发计划(2021YFC2301801)。

摘　　要：目的探究精氨酸代谢在甲型流感病毒(influenza A virus,FluA)感染引起的炎症反应中的作用。方法采用FluA滴鼻感染18月龄老年小鼠,于感染后第6天采集肺组织和支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)。采用Luminex多因子检测法分析BALF中细胞因子水平,采用靶向代谢组学的方法分析BALF中代谢物的变化情况,并对细胞因子与代谢物之间的相关性进行分析。以FluA感染复数(multiplicity of infection,MOI)为1的剂量感染巨噬细胞,并加入不同浓度的精氨酸培养24 h。分别采用实时荧光定量RT-PCR法(real time fluorescence quantitative RT-PCR,qRT-PCR)和流式微球技术(cytometric bead array,CBA)检测白介素(interleukin,IL)-1β、IL-6、肿瘤坏死因子α(tumor necrosis factorα,TNF-α)和IL-10的mRNA和蛋白表达量。结果与对照组相比,FluA感染组老年小鼠BALF中肺表面活性蛋白D(surfactant protein D,SP-D)、TNF-α、单核细胞趋化蛋白1(monocyte chemotactic protein-1,MCP-1)、IL-1α、IL-6、IL-10、S100钙结合蛋白(recombinant S100 calcium binding protein,S100)A9、干扰素诱导蛋白10(interferon inducible protein 10,IP-10)、巨噬细胞炎症蛋白1α(macrophage inflammatory protein-1α,MIP-1α)、基质金属蛋白酶9(matrix metalloprotein-9,MMP9)、补体因子D的水平显著升高,精氨酸代谢通路中精氨酸、天冬氨酸、瓜氨酸、谷氨酸、鸟氨酸、脯氨酸、肌酸和肌氨酸水平上调,且大部分升高的细胞因子水平与精氨酸相关代谢物呈显著正相关。FluA感染后加入精氨酸可以使巨噬细胞中IL-1β、IL-6、TNF-α的表达水平显著降低,而IL-10的水平升高。结论精氨酸可以降低FluA感染诱导的巨噬细胞炎症反应,提示其可能成为FluA感染引起的肺部重症炎症的潜在干预靶点。ObjectiveTo explore the role of arginine metabolism in the inflammatory response to influenza A virus(FluA)infection.MethodsEighteen-month-old mice were infected with FluA via nasal drip,with samples collected on the 6th day post-infection.The concentration of cytokines was determined by the Luminex multifactor assay,while the metabolites in bronchoalveolar lavage fluid(BALF)were analyzed using targeted metabolomic method.Correlation analysis was employed to examine the correlation between cytokines and metabolites.Macrophages were infected with FluA at a multiplicity of infection(MOI)of 1 and cultured with different concentrations of arginine for 24 h.The mRNA and protein expression levels of interleukin(IL)-1β,IL-6,tumor necrosis factorα(TNF-α)and IL-10 were detected by real time fluorescence quantitative RT-PCR(qRT-PCR)and Cytometric Bead Array(CBA).ResultsIn comparison to the control group,the levels of surfactant protein D(SP-D),TNF-α,monocyte chemotactic protein-1(MCP-1),IL-1α,IL-6,IL-10,recombinant S100 calcium binding protein(S100)A9,interferon inducible protein 10(IP-10),macrophage inflammatory protein-1α(MIP-1α),matrix metalloprotein-9(MMP-9),and Complement Factor D in BALF of FluA infection exhibited a significant elevation.The concentrations of arginine,aspartate,citrulline,glutamic acid,ornithine,proline,creatine,and sarcosine in arginine metabolism were up-regulated,which was correlated with most of elevated cytokine levels.The supplementation of arginine after FluA infection significantly decreased the levels of IL-1β,IL-6,and TNF-α,but increased the level of IL-10 in macrophages.ConclusionsArginine reduces the inflammatory response induced by FluA infection in macrophages,suggesting that it may be a potential intervention target for severe pulmonary inflammation following FluA infection.

关 键 词：甲型流感病毒 支气管肺泡灌洗液 细胞因子 精氨酸代谢 

分 类 号：R511.7[医药卫生—内科学]