基于用GFP标记的大肠杆菌构建大鼠可视化细菌性前列腺感染模型

摘要

目的 采用绿色荧光蛋白(green fluorescent protein,GFP)标记的大肠杆菌构建大鼠细菌性前列腺炎模型,观察细菌在组织中的侵袭路径和机体免疫反应,并建立感染菌在体直接定量的分析方法.方法 于分组大鼠左侧前列腺背叶注入大肠杆菌K88-GFP液(1.49×109cfu·ml-1)0.05 ml,采用腺体表面荧光成像及组织切片荧光显微镜观察,研究荧光菌的感染侵袭过程以及机体的免疫反应,建立可视化细菌性前列腺炎模型.结果 ① 通过表面拍照和病理切片得出细菌在腺体内主要侵袭途径,以及组织的病理变化过程.②测定从感染腺体内提取分离的GFP荧光强度,初步建立直接测定感染菌数量的方法.结论 利用GFP标记的细菌可以构建可视化细菌性前列腺炎模型,利用荧光成像技术可以标定感染的程度并进行直接细菌计量,为进一步开展抗菌药PK/PD研究提供新思路.%Aim To induce an experimental model in rats by E. coli tagged with GFP to investigate the significance of the invasion pathway of bacteria and the immune response in abacterial prostatitis and to establish a direct quantitative analysis method of pathogen. Methods After injecting 0. 05 ml of of E. coli K88-GFP ( 1. 49 × 109cfu · ml-1 ) liquid into the left dorsal lobe of SD rat prostate, a visualized method of bacterial prostatitis model was established through surface fluorescence image and tissue sections observed by fluorescence microscopy. Results ① The pathway of invasion and infection and the pathology changes of gland were confirmed. ② A preliminary direct quantitative analysis method of pathogen was established. Conclusion Through constructing a visualized method of bacterial prostatitis model tagged with GFP, a direct quantitative analysis method of pathogen would be established and it provides a novel approach to further study of PK/PD for antibacterial .