孕期流感病毒感染对后代小鼠肺表面活性蛋白表达的影响  

The effect of maternal influenza virus infection during late gestation on the lungs of postnatal Balb/c mice

作　　者：张钰[1] 韦红[1] 周剑芳[2] 于在江[2] 舒跃龙[2] 

机构地区：[1]重庆医科大学附属儿童医院新生儿诊治中心儿童发育疾病研究省部共建教育部重点实验室,400014 [2]中国疾病预防控制中心病毒病预防控制所病毒基因工程国家重点实验室,北京100052

出　　处：《免疫学杂志》2013年第5期422-425,433,共5页Immunological Journal

基　　金：2011年国家临床重点专科建设项目(重庆医科大学附属儿童医院新生儿专业);国家973课题(2011CB504704)

摘　　要：目的研究孕期流感病毒感染对后代小鼠肺组织形态和肺表面活性蛋白(surfactant protein,SP)-A、-D表达的影响,探讨影响机制。方法用非致死量甲型H1N1流感病毒A/四川/SWL1/2009 H1N1稀释液滴鼻感染孕15 d Balb/c孕鼠,在孕19 d随机选取孕鼠剖宫取其胎盘组织,并收集羊水标本;取胎鼠肺组织。待余下孕鼠自然分娩后,取P0,P4,P7,P14 d小鼠肺组织。观察后代小鼠肺组织病理学改变;检测其肺组织SP-A、-D的表达水平。检测胎盘、胎鼠肺组织流感病毒基因;检测羊水白介素-6(interleukin-6,IL-6),肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平。结果与对照组相比,感染组后代小鼠肺泡发育程度较低,肺泡隔厚度明显增加。SP-A的表达量从P4 d开始显著降低;SP-D的表达量从P7 d开始显著升高。未检测到胎盘或胎鼠肺组织流感病毒基因;羊水中IL-6,TNF-α水平明显升高。结论孕期流感病毒感染可能通过造成宫炎症环境间接改变后代肺组织形态和SP-A,-D的表达模式。Evidence from previous influenza pandemics and interpandemic seasonal influenza suggests that pregnant women are at increased risk for influenza complications. But the effect of maternal influenza on the fetus is not as clearly defined. In this study, we aimed to investigate the effect of maternal influenza virus infection during late gestation on postnatal lung surfactant proteins （SP）-A, -D expression in Balb/c mice and explore its potential mechanism. Pregnant Balb/c mice at the fifteenth day of pregnancy （El5） were intranasally inoculated with a sublethal dose of influenza MSichuan/SWL1/2009 （H1N1, SC/1） or sham-infected using vehicle solution after anesthesia with 25 μl isoflurane. At El9, placentas and fetal lungs were harvested, amniotic fluid samples were collected. The presence of H1N1 viral-specific genes for M1 in placentas of infected mice and lungs of exposed offsprings were detected by real time fluorescent quantitative PCR; IL-6 and TNF-cx concentration in amniotic fluid were determined by enzyme-linked immunosorbent assay （ELISA）. On postnatal day 1 （P0）, P4, P7, P14, pathomorphology of the offspring lungs was observed with hematoxylin-eosine staining, while the expression of SP-A and SP-D were detected by Western blotting. Compared with the sham-infected group, the histopathology of exposed offspring lung tissues showed focal alveolar septal widening with pneumocyte hyperplasia, and Western blot demonstrated a decreased expression of SP-A form P4 but a increased expression of SP-D from P7. Furthermore, the concentrations of IL-6 and TNF-α in amniotic fluid were significantly higher, but H 1N 1 viral genes were not detected in placenta or lungs of offspring following infection. Our results demonstrated that maternal influenza virus infection alters the expression of SP-A and SP-D, which implies that the effects of maternal infection on fetal lung development are likely to be indirect.

关 键 词：流感病毒 肺表面活性蛋白 IL-6 TNF-α 

分 类 号：R714.7[医药卫生—妇产科学]