小鼠感染猪致病性大肠杆菌模型的构建与病理研究  

Construction and Pathological Study of Mice Infected with Porcine Pathogenic Escherichia coli

作　　者：王菲 姚美慧 贾雨康 孙江涛 董思言 吕博 曹胜亮 滕建 谢理 李玉保[1] 任庆海 WANG Fei;YAO Mei-hui;JIA Yu-kang;SUN Jiang-tao;DONG Si-yan;LV Bo;CAO Sheng-liang;TENG Jian;XIE Li;LI Yu-bao;REN Qing-hai(College of Agriculture and Biology/Liaocheng University,Liaocheng 252000,China;Tian Shai(Liaocheng)Biotechnology Co.,LTD,Liaocheng 252000,China;Shanchuan Biotechnology(Wuhan)Co.,LTD,Wuhan 430000,China)

机构地区：[1]聊城大学农业与生物学院,山东聊城252000 [2]天筛(聊城)生物科技有限公司,山东聊城252000 [3]山川生物科技(武汉)有限公司,湖北武汉430000

出　　处：《山东农业大学学报(自然科学版)》2025年第2期266-274,共9页Journal of Shandong Agricultural University:Natural Science Edition

基　　金：国家自然科学基金面上项目(32372957)。

摘　　要：猪致病性大肠杆菌是一种能引起猪肠道疾病、败血症和尿路感染的重要病原菌,为探究致病性大肠杆菌诱导的损伤机制,将50只8~9周龄Babl/c的雌性小鼠随机分为10组,每组5只,4个模型组分别腹腔注射1×10~6 cfu/mL、1×10~7 cfu/mL、1×10~8 cfu/mL、1×10~9 cfu/mL大肠杆菌300μL,对照组小鼠分别注射相同体积的灭菌PBS,观察72 h,对临床病理变化、白细胞介素-2(IL-2)、白细胞介素-6(IL-6)的含量进行了观察和检测。通过电击转化法将表达绿色荧光蛋白(GFP)的原核表达载体转化到致病性大肠杆菌EC2316中,获得表达绿色荧光的菌株EC2316-GFP。结果表明:菌株EC2316的半数致死量为2×10~7 cfu/mL,细菌在器官定植随着细菌浓度增加而增加,IL2、IL-6显著增加。导入绿荧光蛋白前后菌株EC2316的生长特性没有改变。综上,小鼠在感染细菌后表现出一系列病理变化及炎性因子变化与自然发病的猪致病性大肠杆菌典型病例相似,成功构建了小鼠感染猪致病性大肠杆菌的模型,为深入研究大肠杆菌引发的损伤机制提供了理论和实践依据。Porcine pathogenic Escherichia coli,is an important pathogen that causes intestinal disease,septicaemia and urinary tract infections.In order to explore the injury mechanism induced by pathogenic Escherichia coli,this experiment randomly divides 508-to 9-week-old Babl/c female mice into 10 groups,with 5 mice in each group.Four model groups are intraperitoneally injected with 300μL E.coli at concentrations of 1×106 cfu/mL,1×107 cfu/mL,1×108 cfu/mL,1×109 cfu/mL respectively,while the control group mice are intraperitoneally injected with an equal volume of sterilized PBS.After 72 hours of observation,the histopathological changes are observed and the cellular inflammatory factors interleukin-2(IL-2)and interleukin-6(IL-6)are detected.The prokaryotic expression vector expressing green fluorescent protein(GFP)is transformed into pathogenic E.coli EC2316 by electroporation transformation method,and the strain EC2316-GFP expressing green fluorescence is obtained.The results show that the median lethal dose of strain EC2316 is 2×107 cfu/mL.The bacterial colonization in organs increases with rising bacterial concentration,and IL2 and IL-6 increased significantly.The introduction of GFP did not alters the growth characteristics of strain EC2316.In summary,the pathological changes and inflammatory responses observed in mice infected with porcine pathogenic E.coli closely resemble those documented in naturally occurring cases.A mouse model of porcine pathogenic E.coli infection is successfully established,which provides theoretical and practical foundation for further research into the damage mechanisms caused by E.coli.

关 键 词：猪致病性大肠杆菌 绿荧光蛋白 LD50 小鼠模型 

分 类 号：S855.12[农业科学—临床兽医学]