小鼠克雷白杆菌肺炎模型的建立及检测  

Establishment and detection of a mouse pneumonia model induced by Klebsiella pneumoniae

作　　者：魏晶[1] 陈纪飞 李强[2] 安英[2] 李芳[1] 

机构地区：[1]大连医科大学基础医学院免疫学教研室,大连116044 [2]吉林医药学院,吉林132013

出　　处：《中国免疫学杂志》2014年第1期85-88,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.30772023)

摘　　要：目的:研究有效建立小鼠克雷白杆菌肺炎模型的方法并进行相关炎症因子检测。方法:构建动物模型,C57小鼠随机分为对照组和模型组,应用气管内注射肺炎克雷白杆菌(106cfu,20#l)的方法建立肺炎模型,感染后24 h,观察小鼠一般状态;收集肺泡灌洗液、分离肺组织,大体观察;肺组织切片HE染色观察两组小鼠炎症情况;肺泡灌洗液进行中性粒细胞计数,肺泡灌洗液和肺组织MPO检测的方法来比较两组小鼠中性粒细胞浸润情况;应用实时定量PCR、Western blot对两组小鼠相关炎症因子进行检测。结果:和对照组相比,模型组小鼠出现明显喘息,寒颤以及竖毛;肺部出现明显的充血、水肿;HE染色显示模型组肺泡壁增厚,大量炎症细胞浸润;模型组肺泡灌洗液中性粒细胞的数量明显高于对照组(P<0.05);模型组支气管肺泡灌洗液和肺组织中MPO表达水平均明显高于对照组(P<0.05);实时定量PCR、Western blot结果均表明模型组小鼠肺组织中炎症因子较对照组明显升高。结论:利用气管内注菌建立小鼠克雷白肺炎模型的方法确实可靠,该模型可用于细胞因子及其受体与克雷白杆菌肺炎关系的研究。Objective：To establish the Klebsiella pneumonia model of mice and to detect inflammatory cytokines. Methods： C57 mice were divided into different groups randomly. The pneumonia models were established by intratracheal Klebsiella pneumoniae （106cfu,20μl）. 24 h after infection, the general status and change in breathing were observed. General observation and HE staining of the lungs were conducted after harvesting. The number of neutrophils in BALF was counted, MPO activity in BALF and lung tissues was detected. The inflammatory cytokines were detected by real time PCR and Western blot. Results： Compared with control group, there were obvious congestion and edema in the lung tissue, more inflammatory cells infiltration after HE staining, more neutrophils in BALF （ P 〈 0.05 ）, and more expression of MPO in the BALF and in the lung tissue in the infected group （ P 〈 0.05 ）. The levels of some inflammatory cytokines were much higher than those in the control group by the detections of qPCR and Western blot. Conclusion： Klebsiella pneumonia model can be established successfully by intratraeheal of Klebsiella pneumonine. This model can be used in the research of the relationship among Klebsiella pneumonia, cytokines and their receptors.

关 键 词：克雷白杆菌 肺炎 模型 

分 类 号：R392[医药卫生—免疫学]