鼠李糖乳杆菌培养上清液通过抑制NF-κB通路预防大肠杆菌性脑膜炎

发布时间：2018-01-24 17:56

本文关键词： 鼠李糖乳杆菌上清液脑膜炎人脑血管内皮细胞 E.coli K NF-κB　出处：《南方医科大学学报》2017年01期 　论文类型：期刊论文

【摘要】：目的体外探讨鼠李糖乳杆菌上清液(LGG-CM)能否通过抑制NF-κB通路来阻断大肠杆菌K1(E.coli K1)株引起的细菌性脑膜炎。方法用人脑微血管内皮细胞(HBMEC)构建体外血脑屏障模型;采用免疫印迹研究LGG-CM能否抑制E.coli K1激活NF-κB通路;通过侵袭实验和中性粒细胞迁移实验,研究LGG-CM能否抑制细菌侵袭和中性粒细胞迁移;通过免疫印迹研究黏附分子CD44和紧密连接蛋白ZO-1的表达;免疫荧光检测ZO-1蛋白的细胞内分布;用Transwell建立体外血脑屏障模型,通过跨细胞内皮电阻(TEER)值和细菌迁移实验评价LGG-CM对细胞屏障完整性的保护作用。结果免疫印迹结果表明LGGCM能抑制E.coli K1激活NF-κB通路,藉此抑制E.coli K1的侵袭和中性粒细胞迁移。同时,LGG-CM可抑制E.coli K1上调CD44蛋白和下调紧密连接蛋白ZO-1。此外,LGG-CM能够明显减缓TEER值的降低和抑制E.coli K1穿越体外血脑屏障。结论体外实验表明,LGG-CM能够通过抑制NF-κB通路激活、阻断E.coli K1侵袭和中性粒细胞迁移及维护血脑屏障完整性来预防E.coli K1引起的细菌性脑炎。
[Abstract]:Objective to investigate whether Lactobacillus rhamnoides supernatant LGG-CM) can block Escherichia coli K1 by inhibiting NF- 魏 B pathway. Methods Human brain microvascular endothelial cells (HBMECs) were used to construct the blood-brain barrier model in vitro. Western blot was used to study whether LGG-CM could inhibit the activation of NF- 魏 B pathway by E. coli K1. To study whether LGG-CM can inhibit bacterial invasion and neutrophil migration through invasion test and neutrophil migration experiment. The expression of adhesion molecule CD44 and tight junction protein ZO-1 was studied by Western blot. The intracellular distribution of ZO-1 protein was detected by immunofluorescence. The model of blood-brain barrier in vitro was established by Transwell. Transcellular endothelial resistance (TEER). Results Immunoblotting results showed that LGGCM could inhibit the activation of NF- 魏 B pathway by E. coli K1. In order to inhibit the invasion of E. coli K1 and neutrophil migration. LGG-CM could inhibit the up-regulation of CD44 protein and down-regulation of tight junction protein ZO-1.In addition, E. coli K1 was inhibited by E. coli K1. LGG-CM could significantly slow down the decrease of TEER and inhibit E.coli K1 from crossing the blood-brain barrier in vitro. LGG-CM can be activated by inhibiting NF- 魏 B pathway. To prevent bacterial encephalitis caused by E. coli K1 by blocking the invasion of E. coli K1 and neutrophil migration and maintaining the integrity of blood-brain barrier.
【作者单位】：南方医科大学公共卫生学院//广东省热带病研究重点实验室微生物学系;南方医科大学第一临床医学院;南加州大学洛杉矶儿童医院Saban研究所;
【基金】：国家自然科学基金(81370740) 南方医科大学公卫学院院长基金(GW201601)~~
【分类号】：R96
【正文快照】： 大肠杆菌E.coli K1是引起新生儿菌血症和脑膜炎最常见的革兰阴性菌[1]。最近的研究表明E.coli K1在侵袭血脑屏障并引发脑膜炎的过程中有着典型的三联症特征,即NF-κB信号通路激活、细菌侵袭脑微血管内皮细胞(BMEC)以及中性粒细胞迁移穿越血脑屏障[2-3]。E.coli K1引起的三联炎

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