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致肾盂肾炎大肠埃希菌(uropathogenic Escherichia coli,UPEC)与尿路上皮细胞之间的相互作用对阐明泌尿道感染的发病机制具有重要意义.本研究利用UPEC132感染非洲绿猴肾细胞(Vero细胞)、人肾癌细胞(Ketr-3细胞)和膀胱癌细胞(EJ细胞),比较细菌对3种细胞的黏附和侵袭能力.结果表明,UPEC132对上述3种细胞均能黏附与侵袭,其黏附率分别为(49.20±7.55)%,(55.22±4.09)%和(73.20±5.26)%,侵袭指数分别为(2.61±0.32)×10-3,(3.00±0.34)×10-3和(3.25±0.20)×10-3.UPEC132对EJ细胞的黏附率高于另外2种细胞(P<0.05),对EJ细胞的侵袭指数与Ketr-3细胞无显著性差异(P>0.05),但高于Vero细胞(P<0.05).用间接免疫荧光法检测细胞表面P血型抗原物质,表明3种细胞均存在P菌毛受体,以EJ细胞表面的受体分布最多.激光共聚焦显微镜可直接观察到表达绿色荧光蛋白的UPEC132/pSELECT-GFP对EJ细胞的侵袭.由于UPEC132对EJ细胞的黏附能力最强,并直接证明具有侵袭性,因此选择EJ细胞作为研究对象为进一步研究UPEC致病机制奠定基础.
The interaction between uropathogenic Escherichia coli (UPEC) and urothelial cells plays an important role in elucidating the pathogenesis of urinary tract infection.In this study, UPEC132 was used to infect Vero cells in African green monkey kidney cells (Vero cells) , Human renal cancer cells (Ketr-3 cells) and bladder cancer cells (EJ cells), and compared the adhesion and invasion ability of the bacteria to the three kinds of cells.The results showed that UPEC132 could adhere and invade the three kinds of cells, (49.20 ± 7.55)%, (55.22 ± 4.09)% and (73.20 ± 5.26)%, respectively. The invasion indexes were (2.61 ± 0.32) × 10-3, (3.00 ± 0.34) × 10-3 and (3.25 ± 0.20) × 10-3.The adhesion rate of UPEC132 to EJ cells was higher than that of the other 2 cells (P <0.05), while the invasion index of EJ cells was not significantly different from that of Ketr-3 cells (P> 0.05) Vero cells (P <0.05) .Indirect immunofluorescence assay of P antigen on the surface of the cells, indicating that P cells are present in all three kinds of cells receptors, the EJ cell surface receptor distribution most. Confocal laser microscopy can be directly observed UPEC132 / pSELECT-GFP expressing green fluorescent protein on EJ cells.Because UPEC132 has the strongest adhesion ability to EJ cells, And directly proved to be invasive, so the selection of EJ cells as the research object for further study of pathogenesis of UPEC lays the foundation.