Enteroglial-derived S100B protein integrates bacteria-induced Toll-like receptor signalling in human enteric glial cells
Enteroglial-derived S100B protein integrates bacteria-induced Toll-like receptor signalling in human enteric glial cells
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DOI：
10.1136/gutjnl-2012-302090
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发表时间：
2014-01-01
期刊：
影响因子：
24.5
通讯作者：
Cuomo, Rosario
中科院分区：
文献类型：
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作者：
Turco, Fabio;Sarnelli, Giovanni;Cuomo, Rosario
目的肠神经胶质细胞（EGC）参与宿主-细菌间的相互作用，在肠道内发挥保护作用。EGC与微生物相互作用的方式仍然知之甚少。我们旨在评估：EGC参与宿主-细菌相互作用; S100 B和Toll样受体（TLR）信号传导会聚在一个共同的途径中，导致一氧化氮（NO）production.Design人EGC的原代培养物暴露于致病性（肠侵袭性大肠杆菌; EIEC）和益生菌（副干酪乳杆菌F19）细菌。通过评估cFos和主要组织相容性复合体（MHC）II类分子的表达来评估细胞活化。通过实时PCR、荧光显微镜和蛋白质印迹分析，在基线和暴露于细菌后评估EGC中的TLR表达。结果EIEC通过诱导cFos和MHC Ⅱ的表达激活EGC，并通过诱导CFos和MHC Ⅱ的表达激活EGC。EGC在基线时表达TLR。病原体和益生菌差异调节TLR在EGC中的表达。病原体，而不是益生菌，显着诱导S100 B蛋白的过度表达和NO释放从EGC。TLR和S100 B通路的特异性抑制剂预处理废除了细菌诱导的NO释放EGC.Conclusions人EGC与细菌相互作用，并通过不同的TLR表达和NO产生来区分病原体和益生菌。在EGC中，在TLR和S100 B通路的特异性抑制剂的存在下，NO释放受损，这表明存在一种新的共同通路，涉及TLR刺激和S100 B蛋白上调。
Objective Enteric glial cells (EGC) have been suggested to participate in host-bacteria cross-talk, playing a protective role within the gut. The way EGC interact with microorganisms is still poorly understood. We aimed to evaluate whether: EGC participate in host-bacteria interaction; S100B and Toll-like receptor (TLR) signalling converge in a common pathway leading to nitric oxide (NO) production.Design Primary cultures of human EGC were exposed to pathogenic (enteroinvasive Escherichia coli; EIEC) and probiotic (Lactobacillus paracasei F19) bacteria. Cell activation was assessed by evaluating the expression of cFos and major histocompatibility complex (MHC) class II molecules. TLR expression in EGC was evaluated at both baseline and after exposure to bacteria by real-time PCR, fluorescence microscopy and western blot analysis. S100B expression and NO release from EGC, following exposure to bacteria, were measured in the presence or absence of specific TLR and S100B pathway inhibitors.Results EIEC activated EGC by inducing the expression of cFos and MHC II. EGC expressed TLR at baseline. Pathogens and probiotics differentially modulated TLR expression in EGC. Pathogens, but not probiotics, significantly induced S100B protein overexpression and NO release from EGC. Pretreatment with specific inhibitors of TLR and S100B pathways abolished bacterial-induced NO release from EGC.Conclusions Human EGC interact with bacteria and discriminate between pathogens and probiotics via a different TLR expression and NO production. In EGC, NO release is impaired in the presence of specific inhibitors of the TLR and S100B pathways, suggesting the presence of a novel common pathway involving both TLR stimulation and S100B protein upregulation.