Zeng Qing, He Xiao-Long, Xiao Han-Sheng, Du Lei, Li Yu-Jing, Chen Le-Cheng, Tian Hui-Wen, Huang Sheng-He, Cao Hong
Department of Microbiology, School of Public healthy, Southern Medical University, Guangzhou 510515, China.E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2017 Jan 20;37(1):24-29. doi: 10.3969/j.issn.1673-4254.2017.01.05.
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.
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.
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.
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.
研究鼠李糖乳杆菌GG条件培养基(LGG-CM)是否通过抑制核因子-κB(NF-κB)信号通路对体外大肠杆菌K1诱导的神经致病性具有预防作用。
用人脑微血管内皮细胞(HBMECs)构建体外血脑屏障(BBB)模型。采用蛋白质免疫印迹法检测LGG-CM对大肠杆菌激活的NF-κB信号通路的影响。进行侵袭试验和多形核白细胞(PMN)迁移试验,以探讨LGG-CM是否能在体外抑制大肠杆菌穿过血脑屏障的侵袭和PMN迁移。采用蛋白质免疫印迹法和免疫荧光法检测紧密连接蛋白1(ZO-1)和细胞表面抗原CD44的表达。通过测定跨上皮电阻(TEER)的变化和细菌移位来评估血脑屏障的通透性。
LGG-CM预处理可抑制HBMECs中大肠杆菌激活的NF-κB信号通路,降低大肠杆菌K1的侵袭和PMN的迁移。蛋白质免疫印迹法显示,LGG-CM可减轻大肠杆菌诱导的HBMECs中CD44表达上调和ZO-1表达下调。此外,LGG-CM预处理可减轻大肠杆菌K1诱导的TEER降低,并在体外抑制细菌穿过血脑屏障的移位。
LGG-CM可阻断大肠杆菌诱导的NF-κB信号通路激活,从而通过降低大肠杆菌K1侵袭率和PMN迁移来预防大肠杆菌K1诱导的神经致病性。