白细胞介素27在呼吸道合胞病毒致肺部感染中的作用机制研究  

Role of Interleukin-27 in Pulmonary Infection with Respiratory Syncytial Virus

作　　者：何燕超[1] 施天昀 施劲东[1] 梅周芳[1] 钱凌[1] 黄琦慧 揭志军[1] 

机构地区：[1]复旦大学附属上海市第五人民医院呼吸内科,上海200240

出　　处：《中国呼吸与危重监护杂志》2016年第4期356-360,共5页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：国家自然科学基金(编号:81370131);上海市卫生系统优秀学科带头人培养计划(编号:XBR2013078);闵行区自然科学研究课题(编号:2012MHZ019)

摘　　要：目的研究呼吸道合胞病毒(RSV)感染小鼠中白细胞介素27(IL-27)水平的变化,以及IL-27在离体细胞中对RSV病毒复制的影响。方法在动物实验中,取7周龄雌性C57小鼠,分别在RSV感染后的第0、0.5、1、2、4、6、8 d处死小鼠(每组5只),左肺提取RNA,运用Real-time PCR法检测RSV感染不同时间后IL-27p28、IL-27EBI3、RSV-M、干扰素β(IFN-β)mRNA的变化,右肺行HE染色。在细胞实验中,培养人肺腺癌上皮细胞(A549),用不同浓度的重组白细胞介素27(rh IL-27)0、1、10和50 ng/m L刺激培养并感染RSV 24 h,运用Real-time PCR法检测干扰素诱导跨膜蛋白3(IFITM3)、IFN-β、RSV-M mRNA的变化,运用Western blot法检测IFITM3、STAT1/p STAT1蛋白的变化。结果 RSV感染小鼠后第4 d病毒复制达到高峰后逐渐降低,8 d后基本恢复。IFN-βmRNA仅在感染后12 h升高,随后迅速恢复正常。IL-27 p28/EBI3基因在感染后第1 d达高峰后逐渐降低,4 d后基本恢复。在离体试验中,发现10 ng/m L、50 ng/m L浓度的rh IL-27刺激感染的A549细胞可抑制病毒复制,促进IFTIM3基因的表达,并且可以促进STAT1磷酸化。不同浓度的rh IL-27刺激后IFN-βmRNA的表达变化无显著差异。结论在RSV感染小鼠后肺组织中IL-27 mRNA表达增高,而IL-27可能通过促进STAT1磷酸化调节IFITM3蛋白的表达,具有抑制RSV复制的作用。Objective To investigate expression level of interleukin-27（IL-27） and its impact on virus replication in lung after infected with respiratory syncytial virus（ RSV）. Methods In vivo,7-week aged female C57 mice were infected with RSV and sacrificed on day 0,0. 5,1,2,4,6,8（ n = 5）. The left lung was extracted for RNA,and then real-time PCR was used to detect the mRNA levels of IL-27p28,IL-27EBI3,RSV-M protein and interferon-β（ IFN-β）. The right lung was used for HE staining. In vitro,human lung epithelial cells（ A549） were infected with RSV,and stimulated with different concentrations of recombinant human interleukin 27（ rh IL-27） in doses of 0,1ng / m L,10 ng / m L,and 50 ng / m L. After 24 hours,the mRNA expression of interferon induced transmembrane protein 3（ IFITM3）,IFN-β,and RSV-M protein were determined by real-time PCR. IFITM3 and STAT1 / p STAT1 protein expression levels were detected by Western blot. Results The viral load in the lungs of mice peaked on 4 day post infection（DPI）,then gradually decreased,and almost returned to normal on 8 DPI. IFN-β increased transiently 12 hours after infection,and then quickly returned to normal baseline. IL-27 p28 / EBI3 levels peaked on 1 DPI,then gradually decreased to normal on 4 DPI. Stimulating RSV-infected A549 cells with rh IL-27 of 10 ng / m L and 50 ng / m L significantly inhibited viral replication,enhanced IFTIM3 mRNA expression,and induced STAT1 phosphorylation. However,rh IL-27 stimulation did not affect IFN-β mRNA expression. Conclusions The IL-27 mRNA expression increases after RSV infection. The inhibition of IL-27 on RSV replication might be related to up-regulation of STAT1 phosphorylation and IFITM3 protein expression.

关 键 词：白细胞介素27 呼吸道合胞病毒 STAT1 P STAT1 

分 类 号：R563.1[医药卫生—呼吸系统]