小鼠鲍曼不动杆菌肺感染模型的制备

发布时间：2018-08-15 18:30

【摘要】：目的构建小鼠鲍曼不动杆菌(A.baumannii)诱导的肺炎模型,研究A.baumannii致病的分子机制。方法实验分为正常对照组、环磷酰胺预处理组、正常小鼠接种组和免疫缺陷小鼠接种组。利用免疫抑制剂环磷酰胺预处理实验雄性C57BL/6小鼠制备免疫缺陷小鼠模型,使用临床重症监护病房(ICU)分离的A.baumannii制备新鲜菌液(1×108集落形成单位/m L),通过气道接种方法接种至小鼠,分别于接种后6、24、72 h,进行小鼠肺及肺泡灌洗液白细胞和中性粒细胞计数,HE染色观察肺组织炎症变化,ELISA检测试剂盒检测血清粒细胞巨噬细胞集落刺激因子(GM-CSF)、γ干扰素(IFN-γ)、白细胞介素1β(IL-1β)、IL-2、IL-4、IL-5、IL-6、IL-10、IL-12、肿瘤坏死因子α(TNF-α)水平。结果免疫功能正常的小鼠感染A.baumannii72 h后,肺内细菌基本被清除,而免疫抑制剂处理的小鼠接种A.baumannii后,肺及血液中细菌计数持续增加;正常小鼠感染6 h后,体内白细胞计数和中性粒细胞升高,至72 h下降,而经过免疫抑制剂处理的小鼠感染后,肺及血液中白细胞计数和中性粒细胞持续升高;感染72 h后,免疫功能正常小鼠肺部有轻微炎症,免疫缺陷小鼠肺部炎症较为明显;正常小鼠感染6 h后,血清中上述细胞因子含量均增加,至72 h下降,而经过免疫抑制剂处理的小鼠感染后,肺及血液中上述细胞因子水平均持续升高。结论成功制备了A.baumannii诱导的小鼠肺部感染模型。
[Abstract]:Objective to establish a mouse model of pneumonia induced by Acinetobacter baumannii (A.baumannii) and to study the molecular mechanism of A.baumannii. Methods the experiment was divided into normal control group, cyclophosphamide pretreatment group, normal mice inoculation group and immune deficiency mice inoculation group. Immunosuppressant cyclophosphamide was used to pretreat male C57BL/6 mice to establish the model of immunodeficient mice. Fresh bacterial solution was prepared by A.baumannii isolated from (ICU) of clinical intensive care unit (1 脳 10 8 colony forming unit / m L), was inoculated into mice by airway inoculation method). The leukocytes and neutrophils in lung and alveolar lavage fluid of mice were stained with HE to observe the changes of lung inflammation. The serum granulocyte macrophage colony stimulating factor (GM-CSF) and interferon 纬 (IFN- 纬) were detected by Elisa kit. The levels of IL-12, TNF- 伪, TNF- 伪 and IL-6, IL-10, IL-12, TNF- 伪, IL-12, TNF- 伪, IL-12, TNF- 伪, TNF- 伪, IL-12, TNF- 伪, TNF- 伪, TNF- 伪, Results the bacteria in the lungs of mice with normal immune function were basically cleared after infection with A.baumannii72 h, but the counts of bacteria in lung and blood of mice treated with immunosuppressant increased continuously after A.baumannii inoculation, and the bacteria count in normal mice increased after 6 hours of infection. The leukocyte count and neutrophils increased in vivo and decreased at 72 h, but the leukocyte count and neutrophils in lung and blood of mice treated with immunosuppressant increased continuously after 72 h of infection. There was slight inflammation in the lungs of normal immune function mice, but obvious inflammation in the lungs of immunodeficient mice, and the contents of above mentioned cytokines in serum of normal mice increased after 6 h of infection, and decreased at 72 h. The levels of these cytokines in lung and blood of mice treated with immunosuppressant increased continuously after infection. Conclusion A.baumannii induced lung infection in mice was successfully established.
【作者单位】：陕西中医药大学病原微生物及检验教研室;陕西中医药大学第一附属医院检验科;
【基金】：陕西中医药大学自然科学青年基金(2016QN29) 陕西省教育厅专项科研计划(17JK0200)
【分类号】：R-332;R563.1

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