南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (07): 965-.

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ppk1基因缺失对脑膜炎大肠杆菌K1株生物学功能的影响

彭亮,潘嘉韵,罗苏,杨正慧,黄慕芳,曹虹   

  • 出版日期:2014-07-20 发布日期:2014-07-20

Changes of biological behavioral of E. coli K1 after ppk1 gene deletion

  • Online:2014-07-20 Published:2014-07-20

摘要: 目的比较大肠杆菌K1 株E44 敲除ppk1 基因后与野生株之间差异并探讨ppk1 基因在E.coli K1 株致脑膜炎机制中的作
用。方法(1)将野生株与敲除株置于56 ℃ 2、4、6 min,以比较二者对热刺激的抵抗力差异;(2)利用电子显微镜直接观察以及
采用经典的粘附、侵袭率定量实验比较二者对脑微血管内皮细胞(HBMEC)的粘附和侵袭能力;(3)将细菌与脑微血管内皮细胞
共同孵育后,利用激光共聚焦观察二者诱导脑微血管内皮细胞的细胞骨架重排现象。结果与野生株E44相比,ppk1敲除株对
于56 ℃热刺激的抵抗力明显下降;电子显微镜可以观察到,敲除株粘附和侵袭入HBMEC的数量均少于野生株,定量粘附、侵袭
实验也进一步证实;借助激光共聚焦发现敲除株诱导HBMEC细胞骨架重排的能力要弱于野生株。结论ppk1对于脑膜炎大肠
杆菌K1株抵抗热刺激、粘附和侵袭HBMEC以及诱导HBMEC的细胞骨架重排具有重要作用。

Abstract: Objective To study the changes in biological behaviors of meningitis E. coli K1 strain E44 after deletion of
polyphosphate kinase 1 (ppk1) gene and explore the role of ppk1 in the pathogenesis of E. coli K1-induced meningitis. Methods
The wild-type strain E. coli K1 and ppk1 deletion mutant were exposed to heat at 56 ℃ for 6 min, and their survival rates were
determined. The adhesion and invasion of the bacteria to human brain microvascular endothelial cells (HBMECs) were
observed using electron microscopy and quantitative tests. HBMECs were co-incubated with wild-type strain or ppk1 deletion
mutant, and the cytoskeleton rearrangement was observed under laser scanning confocal microscope. Results The survival
rate of the ppk1 deletion mutant was significantly lower than that of the wild-type strain after heat exposure. The ppk1 deletion
mutant also showed lowered cell adhesion and invasion abilities and weakened ability to induce cytoskeleton rearrangement
in HBMECs. Conclusion ppk1 gene is important for E.coli K1 for heat resistance, cell adhesion and invasion, and for inducing
cytoskeletal rearrangement in HBMECs.