南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2014年
7期
965-968
,共4页
彭亮%潘嘉韵%罗苏%杨正慧%黄慕芳%曹虹
彭亮%潘嘉韻%囉囌%楊正慧%黃慕芳%曹虹
팽량%반가운%라소%양정혜%황모방%조홍
脑膜炎%聚磷酸盐激酶1%大肠杆菌K1株%基因敲除
腦膜炎%聚燐痠鹽激酶1%大腸桿菌K1株%基因敲除
뇌막염%취린산염격매1%대장간균K1주%기인고제
meningitis%polyphosphate kinase%Escherichia coli%gene deletion
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。
目的:比較大腸桿菌K1株E44敲除ppk1基因後與野生株之間差異併探討ppk1基因在E.coli K1株緻腦膜炎機製中的作用。方法(1)將野生株與敲除株置于56℃2、4、6 min,以比較二者對熱刺激的牴抗力差異;(2)利用電子顯微鏡直接觀察以及採用經典的粘附、侵襲率定量實驗比較二者對腦微血管內皮細胞(HBMEC)的粘附和侵襲能力;(3)將細菌與腦微血管內皮細胞共同孵育後,利用激光共聚焦觀察二者誘導腦微血管內皮細胞的細胞骨架重排現象。結果與野生株E44相比,ppk1敲除株對于56℃熱刺激的牴抗力明顯下降;電子顯微鏡可以觀察到,敲除株粘附和侵襲入HBMEC的數量均少于野生株,定量粘附、侵襲實驗也進一步證實;藉助激光共聚焦髮現敲除株誘導HBMEC細胞骨架重排的能力要弱于野生株。結論 ppk1對于腦膜炎大腸桿菌K1株牴抗熱刺激、粘附和侵襲HBMEC以及誘導HBMEC的細胞骨架重排具有重要作用。
목적:비교대장간균K1주E44고제ppk1기인후여야생주지간차이병탐토ppk1기인재E.coli K1주치뇌막염궤제중적작용。방법(1)장야생주여고제주치우56℃2、4、6 min,이비교이자대열자격적저항력차이;(2)이용전자현미경직접관찰이급채용경전적점부、침습솔정량실험비교이자대뇌미혈관내피세포(HBMEC)적점부화침습능력;(3)장세균여뇌미혈관내피세포공동부육후,이용격광공취초관찰이자유도뇌미혈관내피세포적세포골가중배현상。결과여야생주E44상비,ppk1고제주대우56℃열자격적저항력명현하강;전자현미경가이관찰도,고제주점부화침습입HBMEC적수량균소우야생주,정량점부、침습실험야진일보증실;차조격광공취초발현고제주유도HBMEC세포골가중배적능력요약우야생주。결론 ppk1대우뇌막염대장간균K1주저항열자격、점부화침습HBMEC이급유도HBMEC적세포골가중배구유중요작용。
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.