耳窥镜直视下气管插管法构建小鼠新型隐球菌肺炎合并脑膜炎模型  

Establishment of a Cryptococcus neoformans murine pneumonia combined with meningitis model by otoscope guiding intubation method

作　　者：李鑫 赵旭 郑建铭 李谦 Li Xin;Zhao Xu;Zheng Jianming;Li Qian(Institute of Antibiotics,Huashan Hospital,Fudan Universit;Key Laboratory of Clinical Pharmacology of Antibiotics,Ministry of Health,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院抗生素研究所卫生部抗生素临床药理重点实验室,上海200040 [2]复旦大学附属华山医院感染科,上海200040

出　　处：《中华传染病杂志》2018年第5期291-295,共5页Chinese Journal of Infectious Diseases

基　　金：上海市科委实验动物研究专项基金（15140903300）

摘　　要：目的采用耳窥镜直视下气管插管法构建小鼠新型隐球菌肺炎合并脑膜炎模型。方法20只雄性C57BL/C小鼠分为正常对照组和感染组，每组均为10只，实验当日小鼠在耳窥镜直视下行气管插管后，感染组沿气管内插管注射10 μL新型隐球菌，正常对照组沿气管内插管注射10 μL氯化钠溶液，在注射细菌后的0 h、第3、5、7和10天观察小鼠一般情况和体质量，于0 h每组处死3只小鼠，第10天处死剩余小鼠，进行小鼠肺脏、肝脏、脾脏和脑组织等组织菌落计数和肺、脑组织病理检查以判断肺炎合并脑膜炎模型是否构建成功。结果感染组10只小鼠行耳窥镜直视下气管插管，均获成功，无小鼠插管后死亡。沿气管内插管注射菌液浓度为2.0×10^9菌落形成单位（colony-forming units, CFU）/mL，气管插管后0 h处死小鼠取肺并定量，其菌落计数为（7.84±0.13） lg CFU/g。2组小鼠观察至感染后10 d，均无小鼠死亡。第10天小鼠肺内菌量平均值达到（8.11±0.81） lg CFU/g，与0 h比较略有增加，第10天小鼠脑组织内菌量达到（4.84±0.21） lg CFU/g。病理结果提示支气管、肺泡及间质内见大量隐球菌，脑实质内见灶性隐球菌浸润。结论采用耳窥镜直视下气管插管法成功构建了小鼠新型隐球菌肺炎合并脑膜炎模型，方法操作简便，成功率高。ObjectiveTo establish a Cryptococcus neoformans murine pneumonia combined with meningitis model by otoscope guiding intubation method.MethodsTwenty male C57BL/C mice were randomized into two groups, including control group and infection group, with 10 mice in each group. On the experiment day, the mice were intubated by otoscope guiding. The mice in infection group were inoculated with 10 μL of fresh Cryptococcus neoformans suspension through endotracheal tube, while the mice in control group were inoculated with 10 μL of sterile saline. The general conditions were observed and the body masses were measured at 0 h, day 3, day 5, day 7 and day 10 after inoculation in each group. Three mice in each group were sacrificed at 0 h and the rest were sacrificed at day 10. The lungs, livers, spleen and brains from each mouse were aseptically collected for quantitative culture and histopathology to confirm the successful establishment of mouse model.ResultsTen mice were all successfully intubated by otoscope guiding and none of the mice was dead. The concentration of inoculated bacterial solution was 2.0×109 colony-forming units （CFU）/mL. The mice were sacrificed at 0h after incubation and the lungs were collected. The pulmonary bacterial culture at baseline （0 h） was （7.84±0.13） lg CFU/g lung. No mice was dead in two groups from 0 h to day 10. The average pulmonary bacterial load in lung was （8.11±0.81） lg CFU/g at day 10, which increased 16 times comparing with baseline. The average cerebral bacterial load in brain was （4.84±0.21） lg CFU/g at day 10. The histopathologic changes in lung showed lots of Cryptococcus neoformans which invaded in the bronchi, alveoli and alveolar space. The pathology in brain showed focal Cryptococcus neoformans invasion.ConclusionsEstablishment of Cryptococcus neoformans murine pneumonia and meningitis model using otoscope guiding intubation method is successful. This method has the advantages of easy operation and high successful rate.

关 键 词：隐球菌 新型 耳镜 插管法 气管内 肺炎 脑膜炎 模型 动物 

分 类 号：R-332[医药卫生] R519.4