流感病毒感染后接种肺炎链球菌继发中耳炎小鼠模型的建立  

Establishment of mouse model of secondary otitis media by trans-bullar injection of Streptococcus pneumoniae following influenza infection

作　　者：范芳梅 马毓蓉 王子萌[1] 王维[1] 董益麟 何纤 张欣欣 何於娟[1] 

机构地区：[1]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆400016

出　　处：《第三军医大学学报》2018年第5期400-405,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81373151)~~

摘　　要：目的建立甲型流感病毒(influenza A viruses,IAV)感染后继发肺炎链球菌(Streptococcus pneumoniae,Spn)感染性中耳炎小鼠模型。方法经鼻感染不同剂量甲型H1N1型流感病毒PR8,感染后第5天听泡穿刺接种Spn建立C57BL/6小鼠继发感染中耳炎模型,观察小鼠体质量变化和生存率;于建模后不同时间点处死小鼠,制备中耳组织切片HE染色,收集中耳灌洗液进行细胞及细菌计数,ELISA检测灌洗液中炎症因子。结果 PR8预感染可导致Spn感染性中耳炎小鼠死亡,而单纯Spn感染小鼠无死亡,小鼠体质量变化及生存率与病毒剂量呈剂量依赖方式;PR8感染后继发Spn感染小鼠中耳腔炎症细胞浸润明显多于单纯Spn感染小鼠,且细菌载量明显增加;PR8继发Spn感染小鼠中耳腔炎症因子IL-6、IFN-γ与单独Spn感染小鼠差异无统计学意义(P>0.05),而TNF-α明显增高(P<0.05)。结论成功建立了IAV感染后继发Spn感染性中耳炎C57BL/6小鼠模型。Objective To develop a model of secondary otitis media by Streptococcus pneumoniae （Spn） following influenza A virus （IAV） infection in C57BIJ6 mice and explore its pathological mechanism. Methods C57BL/6 mice were infected intranasally with different doses of HI NI （PR8）. In 5 d later, the mice were inoculated with Spn via transbullar injection to induce secondary odds media. Body weight and survival were observed daily in all the mice. Then the mice were killed in different time periods after Sire inoculation. The middle ear （ME） tissues were collected for histological examination by HE staining, and the middle ear lavage fluid （MELF） were harvested for cell counting and bacterial counting. ELISA was used to detect the contents of pro-inflammatory eytokines in the MELF. Results Antecedent PR8 infection led to death in the mice with Spn-indueed otitis media, while simple Spn inoculation had no such effect. The changes of body weight and survival rate were in a dose-dependent manner of PR8 infection. There were more infiltrated inflammation cells and larger bacterial counts in the ME tissue of the model mice when compared with the mice after simple Spn inoculation. There were no significant differences in the production of proinflammatory cytokines IL-6 and IFN-γ among the model mice of secondary otids media （ P 〉 0.05 ） , whereas TNF-α showed a remarkable increase than the mice after simple Spn inoculation （ P 〈 0.05）. Conclusion A model of secondary otitis media of Spn following influenza A virus infection mice is successfully established in C57BL/6 mice.

关 键 词：甲型流感病毒H1N1 继发感染 肺炎链球菌Spn 中耳炎 

分 类 号：R-332[医药卫生] R511.7