致病性大肠杆菌引发宿主细胞A／E损伤分析方法的建立  

Evaluation of A/E lesions in HeLa cells caused by pathogenic E.coli:Giemsa staining and fluorescence actin staining

作　　者：王海光[1] 顾江[1] 余抒[1] 张卫军[1] 邹全明[1] 朱凤才[2] 毛旭虎[1] 

机构地区：[1]第三军医大学医学检验系临床微生物学及免疫学教研室,重庆400038 [2]江苏省疾病预防控制中心,南京210009

出　　处：《第三军医大学学报》2009年第8期659-662,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30670107);国家高技术研究发展计划(863计划)(2006AA02Z443);重庆市自然科学基金(2006BB5115)~~

摘　　要：目的建立一种能够对由致病性大肠杆菌引发的宿主细胞黏附及擦拭性损伤(A/E损伤)进行半定量分析的方法,为进一步研究其致病性提供评价手段。方法以大肠杆菌黏附HeLa细胞作为感染模型。采用姬姆萨染色和荧光肌动蛋白染色实验,通过菌落计数及肌动蛋白聚集数量定量分析,统计学处理,比较EPEC 43095、EHEC O157∶H7 Sakai株和弱毒株EHEC O157∶H7 00B015以及E.coliDH5α对HeLa细胞的黏附能力,评价它们对细胞的损伤程度。结果EPEC对细胞的黏附及擦拭性损伤程度大于EHEC(P<0.05)。EHEC Sakai和00B015在黏附能力上差异不大(P>0.05),但是,Sakai在肌动蛋白聚集形成能力方面却明显强于00B015(P<0.05)。结论成功建立了一种能够对EPEC或EHEC所引发的宿主细胞黏附及擦拭性损伤进行半定量分析的方法。Objective To establish a semi-quantitative method to analyze the attaching and effacing （A/E） lesions on host cells caused by pathogenic Escherichia coli （E. coil）. Methods HeLa cells were infected with enteropathogenic or enterohemorrhagic E. coli strains （EPEC 43095, EHEC O157：H7 Sakai, EHEC O157：H7 00B015 and E. coli DH5α） and visualized by Giemsa staining and fluorescence actin staining （FAS）. Then the numbers of microcolonies and actin polymerizations formed on the cells were counted and calculated. Results All EPEC induced stronger A/E lesions than EHEC on HeLa cells （P 〈 0.05）. The EPEC 43095 caused the strongest A/E lesions among these E. coli strains detected. Although the ability of EHEC Sakai to attach to HeLa cells was similar to that of EHEC 00B015 （P 〉 0.05 ）, EHEC Sakai induced stronger actin polymerizations in host cells than EHEC 00B015 （P 〈0.05）. Conclusion A semi-quantitative method to analyze the A/E lesions of host cells caused by pathogenic E. coli is successfully established. This work contributes to further studies on pathogenicitv of the pathogenic E. coli.

关 键 词：肠致病性大肠杆菌 肠出血性大肠杆菌 黏附和擦拭性损伤 

分 类 号：R-33[医药卫生] R378.21