[1]曾庆,何肖龙,肖汉森,等.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[J].南方医科大学学报,2017,(01):24.
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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()
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《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2017年01期
页码:
24
栏目:
出版日期:
2017-01-20

文章信息/Info

Title:
Lactobacillus rhamnosus GG conditioned medium prevents E. coli meningitis by inhibiting nuclear factor-κB pathway
作者:
曾庆何肖龙肖汉森杜蕾李雨静陈乐程田慧文黄胜和曹虹
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 K1NF-κB
Keywords:
Lactobacillus rhamnosus GG conditioned medium meningitis human brain microvascular endothelial cells E. coli K1 nuclear factor-κB
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 LGG-CM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 rhamnosus GG conditioned medium(LGG-CM)has preventive effect against E. coli K1-induced neuropathogenicity in vitro by inhibiting nuclear factor-κB (NF-κB) signaling pathway. Methods An in vitro blood-brain barrier (BBB) model was constructed using human brain microvascular endothelial cells (HBMECs). The effect of LGG-CM on E. coli-actived NF-κB signaling pathway was assayed using Western blotting. Invasion assay and polymorphonuclear leukocyte (PMN) transmigration assay were performed to explore whether LGG-CM could inhibit E. coli invasion and PMN transmigration across the BBB in vitro. The expressions of ZO-1 and CD44 were detected using Western blotting and immunofluorescence. The changes of trans-epithelial electric resistance (TEER) and bacterial translocation were determined to evaluate the BBB permeability. Results Pre-treament with LGG-CM inhibited E. coli-activated NF-κB signaling pathway in HBMECs and decreased the invasion of E. coli K1 and transmigration of PMN. Western blotting showed that LGG-CM could alleviate E. coli-induced up-regulation of CD44 and down-regulation of ZO-1 expressions in HBMECs. In addition, pre-treatment with LGG-CM alleviated E. coli K1-induced reduction of TEER and suppressed bacterial translocation across the BBB in vitro. Conclusion LGG-CM can block E. coli-induced activation of NF-κB signaling pathway and thereby prevents E. coli K1-induced neuropathogenicity by decreasing E. coli K1 invasion rates and PMN transmigration.

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更新日期/Last Update: 1900-01-01