致肾盂肾炎大肠杆菌对人肾盂上皮原代细胞黏附的实验研究  

Adhesion of uropathogenic Escherichia coli to human renal pelvis epithelial primary cells

作　　者：谷超[1] 陈锦英[1] 侯敏[1] 贺靖冬[1] 畅继武[2] 

机构地区：[1]天津医科大学基础医学院,300070 [2]天津医科大学第二医院

出　　处：《中华微生物学和免疫学杂志》2007年第9期802-806,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金(30470096)

摘　　要：目的建立人肾盂上皮原代细胞的培养方法,用于致肾盂肾炎大肠杆菌(UPEC)黏附作用的研究。方法利用复合PCR方法检测UPEC 132菌株毒力基因hly和cnfl。采集人肾脏手术标本,利用组织块法取肾盂正常上皮组织,使用添加EGF及BPE的角朊细胞无血清培养基(K-SFM)进行原代细胞培养,经HE染色和免疫细胞化学染色进行鉴定。鉴定符合要求的人肾盂上皮原代细胞用于UPEC黏附试验,分别于不同作用时间观察黏附后细胞形态学变化,并计算黏附率和黏附指数。结果此研究制备的人肾盂上皮原代细胞具备移行上皮细胞特征。带有毒力基因hly和cnfl的UPEC 132菌株作用于人肾盂上皮原代细胞,15 min后细菌开始黏附,120min达到高峰,黏附率为74.4%,黏附指数为34.0。显微镜下观察细胞形变、着色不均,胞浆和胞核都有改变。结论人肾盂上皮原代细胞可用于UPEC的黏附试验,为其致病机理的深入研究奠定基础。Objective To establish an in vitro model of primary epithelial cells of human renal pelvis and to study the adhesion of uropathogenie Escherichia coli（UPEC） to them. Methods The virulence genes My and cnfl of UPEC 132 strain were detected by multiplex PCR. Primary cells culture of normal renal pelvis epithelial tissue from human kidney surgery specimen was performed by ＂tissue cubes＂ methods using keratinocyte serum free medium（K-SFM） with EGF and BPE. The primary cells were identified by HE and immunocytochemistry staining. During the UPEC 132 adhesion to primary epithelial cells of human renal pelvis, morphological changes of the adhered cells were observed and the adhesion rates and indexes were recorded at the various experiment times. Results Primary epithelial cells of human renal pelvis possessed the characterizations of the transitional epithelium. When compared with the negative control group, the adhesion rates and indexes began to increase in 15 min and reached peak at 120 min. By microscopy showed significant changes in the adhered primary cells with UPEC 132 strain. Conclusion Primary epithelial cell of human renal pelvis can be used as an in vitro model, to the UPEC adhesion experiment.

关 键 词：致肾盂肾炎大肠杆菌 人肾盂上皮原代细胞 原代培养 黏附 毒力基因 

分 类 号：R378.2[医药卫生—病原生物学]