不同月龄BALB/c小鼠感染甲型H1N1型流感病毒的炎性调控差异研究Regulation of Inflammatory Mediators in BALB/c Mice of Different Ages Infected with the Influenza A H1N1 Virus
孙竹筠;晋一帆;刘孝云;叶冬雪;赵涛;肖敏;容蓉;杨勇;
摘要(Abstract):
为对比不同月龄BALB/c小鼠感染甲型H1N1型流感病毒后的淋巴细胞相对含量和炎症因子调控差异,提供流感类病毒性疾病防控的基础研究证据,本研究选用2月、8月龄雌性BALB/c小鼠,分别感染1×105TCID50/0.1mL的甲型H1N1型流感病毒PR8毒株,每天记录体重、肛温变化,于感染第7d处死解剖取材,计算脏器指数,qPCR技术检测肺组织流感病毒M基因拷贝数,流式细胞术检测全血CD3+、CD4+、CD8+淋巴细胞含量及肺组织中IL-6、IFN-γ、MCP-1等炎症因子的分泌水平。结果显示:(1)感染相同剂量病毒7d后,2月龄小鼠有1例样本死亡,生存质量下降较大,8月龄小鼠全部存活,体重较第0d相比仅下降12.35%,感染后2月龄组小鼠胸腺指数明显下降(P<0.01),8月龄组变化不明显。(2)2月龄组染毒小鼠肺组织M基因拷贝数显著低于8月龄染毒组(P<0.05)。(3)染毒后,2月龄组小鼠体内IFN-γ、IFN-β、IL-6等促炎性细胞因子含量增长显著(P<0.05),IL-6、MCP-1、IL-23含量显著高于8月龄组(P<0.05);2月龄染毒组小鼠杀伤性T淋巴细胞CD8+相对比例也明显高于8月龄染毒组(P<0.05)。可推测2月龄小鼠在感染病原体后产生了较强的免疫反应以清除病毒,但免疫系统的过度反应导致体内IL-6等炎症因子大量分泌,可能出现了细胞因子风暴现象致使机体损伤严重,使2月龄实验组小鼠较8月龄相比出现了更差的状态与较高的死亡率。本研究提示低年龄段人群感染相同剂量的流感病毒时较高年龄段人群可能会出现更严重的病理损伤。
关键词(KeyWords): 甲型H1N1流感病毒;免疫应答;免疫失调;动物模型
基金项目(Foundation): 自主培养创新团队(项目号:2021GXRC028),题目:基于多靶点抗新型冠状病毒抑制剂筛选及高通量免疫调节剂表型发现模型的组分中药研究;; 山东省自然科学基金(项目号:ZR2021LZY012),题目:基于计算机虚拟筛选和高通量靶点筛选相结合的抗新型冠状病毒活性中药发现;; 国家自然科学基金(项目号:81873220),题目:基于证-毒-效关联技术辨识麻黄细辛附子汤对心肌线粒体细胞色素C氧化酶调控的非线性生物学靶向特征~~
作者(Authors): 孙竹筠;晋一帆;刘孝云;叶冬雪;赵涛;肖敏;容蓉;杨勇;
DOI: 10.13242/j.cnki.bingduxuebao.004230
参考文献(References):
- [1]Zhang F,Hu F. The Novel Influenza A(H1N1)[J].Sun Yat-sen Univ,Med Sci,2009,30(5):481-485.
- [2]Liu M,Zhao X,Hua S,Du X,Peng Y,Li X,Lan Y,Wang D,Wu A,Shu Y,Jiang T. Antigenic patterns and evolution of the human influenza A H1N1)Virus[J]. Sci Rep,2015,5(14171).
- [3]Ygberg S,Nilsson A. The developing immune systemfrom foetus to toddler[J]. Acta Paediatr,2012,101(2):120-127.
- [4]Talbot H K. Influenza in Older Adults[J]. Infect Dis Clin North Am,2017,31(4):757-766.
- [5]Dutta S,Sengupta P. Men and mice:Relating their ages[J]. Life Sci,2016,152(244-248).
- [6]Bianconi V,Sahebkar A,Atkin S L,Pirro M. The regulation and importance of monocyte chemoattractant protein-1[J]. Curr Opin Hematol,2018,25(1):44-51.
- [7]Shiomi A, Usui T. Pivotal roles of GM-CSF in autoimmunity and inflammation[J]. Mediators Inflamm,2015,2015568543.
- [8]Schrauwen E J,Herfst S,Chutinimitkul S,Bestebroer T M,Rimmelzwaan G F,Osterhaus A D,Kuiken T,Fouchier R A. Possible increased pathogenicity of pandemic(H1N1)2009 influenza virus upon reassortment[J]. Emerg Infect Dis,2011,17(2):200-208.
- [9]Chowell G,Bertozzi S M,Colchero M A,Lopez-Gatell H,Alpuche-Aranda C,Hernandez M,Miller M A.Severe respiratory disease concurrent with the circulation of H1N1 influenza[J]. N Engl J Med,2009,361(7):674-679.
- [10]Betakova T,Kostrabova A,Lachova V,Turianova L.Cytokines induced during influenza virus infection[J].Curr Pharm Des,2017,23(18):2616-2622.
- [11]Tisoncik J R,Korth M J,Simmons C P,Farrar J,Martin T R,Katze M G. Into the eye of the cytokine storm[J]. Microbiol Mol Biol Rev,2012,76(1):16-32.
- [12]Ryabkova V A,Churilov L P,Shoenfeld Y. Influenza infection,SARS,MERS and COVID-19:Cytokine storm-The common denominator and the lessons to be learned[J]. Clin Immunol,2021,223108652.
- [13]Liu Q,Zhou Y H,Yang Z Q. The cytokine storm of severe influenza and development of immunomodulatory therapy[J]. Cell Mol Immunol,2016,13(1):3-10.
- [14]Garcia M,Misplon J A,Price G E,Lo C,Epstein S L.Age dependence of immunity induced by a candidate universal influenza vaccine in mice[J/OL]. PLoS One,2016,11(4):e0153195.
- [15]Wing L S Y,Loo E X L,Kang A Y H,Lau H X,Tambyah P A,Tham E T. Age-Related differences in immunological responses to SARS-CoV-2[J]. J Allergy Clin Immunol Pract,2020,8(10):3251-3258.
- [16]Teijaro J R. Cytokine storms in infectious diseases[J].Semin Immunopathol,2017,39(5):501-503.