肺炎克雷伯杆菌在T24细胞内存活的动态观察  

An Investigation of the dynamic processes on the survival of Klebsiella pneumoniae in Urinary bladder epithelial cell

作　　者：刘行海[1] 魏桂花[1] 路会侠[1] 熊文碧[1] 黄宁[1] 王伯瑶[1] 吴琦[1] 

机构地区：[1]四川大学华西医学中心感染免疫室,四川成都610044

出　　处：《四川生理科学杂志》2008年第2期49-51,共3页Sichuan Journal of Physiological Sciences

基　　金：国家自然科学基金(NO:30570790);教育部博士点基金资助

摘　　要：目的:了解肺炎克雷伯杆菌与膀胱上皮细胞的相互关系,观察肺炎克雷伯杆菌在人膀胱上皮细胞株T24中生存的动态变化。方法:采用肺炎克雷伯杆菌临床分离株03138侵袭T24细胞,并用庆大霉素杀死细胞外的细菌,分别于细菌进入细胞后的4、24、48及72h裂解细胞,释放出细胞内的活细菌,用平板菌落计数法计数胞内活菌数。结果:T24细胞内的肺炎克雷伯杆菌03138株在实验48h内有一定生长,试验72h细胞内活菌数量明显减少。加入细胞因子(TNF-α和INF-γ)可以促进上皮细胞清除胞内细菌。结论:膀胱上皮细胞清除进入细胞内的肺炎克雷伯杆菌,可能是泌尿道天然免疫的一种防御机制,而细胞因子可以调控上皮细胞的抗菌作用。Objective： To understand the interaction between Klebsiella pneumoniae and urinary epithelial cells, intracellular survival curve of klebsiella pneumoniae in human urinary bladder epithelial cell line T24 was assayed. Methods： Monolayers of T24 cell were incubated with a klebsiella pneumoniae clinical isolate 03183 （approximately 100 bacteria per epithelial cell） for 2 h. T24 cells were lysed with Triton X-100 at 4, 24, 48 and 72 hours after the initial incubation. Viable intracellular bacteria were quantified by plating appropriate dilutions on LB agar plates. Meanwhile the effect of proinflammatory cytokines TNF-α and INF-γ on the survival of klebsiella pneumoniae in urinary bladder epithelial cell was investigated. Results： After 48 h of incubation, there was a 34% increase in the number of viable intracellular klebsiella pneumoniae compared with intracellular bacteria recovered after 4 h of incubation. However, the number of viable intracellular bacteria recovered after 72 h of incubation decreased 66%. In the presence of TNF-αand INF-γ,intracellular viable bacteria decreased 73% compared with the control. Conclusion：The reduction of klebsiella pneumoniae viability over time suggests that urinary bladder epithelial cells have the ability to clear invading Klebsiella pneumoniae which might represent an important host defense mechanism against urinary tract infection.

关 键 词：肺炎克雷伯杆菌 泌尿道上皮细胞 抗菌作用 

分 类 号：R378[医药卫生—病原生物学]