Molecular Signaling and Dysfunction of the Human Reactive Enteric Glial Cell Phenotype: Implications for GI Infection, IBD, POI, Neurological, Motility, and GI Disorders.
Molecular Signaling and Dysfunction of the Human Reactive Enteric Glial Cell Phenotype: Implications for GI Infection, IBD, POI, Neurological, Motility, and GI Disorders.
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DOI：
10.1097/mib.0000000000000854
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发表时间：
2016-08
影响因子：
4.9
通讯作者：
Christofi FL
中科院分区：
文献类型：
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作者：
Liñán-Rico A;Turco F;Ochoa-Cortes F;Harzman A;Needleman BJ;Arsenescu R;Abdel-Rasoul M;Fadda P;Grants I;Whitaker E;Cuomo R;Christofi FL
临床观察或动物研究表明肠神经胶质细胞（EGC）与运动障碍、IBS、IBD、GI感染、术后肠梗阻和慢传输型便秘有关。人类胃肠道中神经胶质对炎症反应的潜在机制尚不清楚。我们的目标是确定炎症诱导的“反应性人类EGC表型”，并探讨其功能相关性。从15个胃肠道手术标本中培养的人EGC被用来研究基因表达，Ca 2+和嘌呤能信号通过Ca 2 +/fluo-4成像和机械敏感性。设计了107个基因的纳米串面板，作为炎症、转录、嘌呤能信号传导、囊泡转运蛋白、通道、抗氧化剂和其他途径的读出。采用脂多糖（LPS，200μg/ml）和干扰素-γ（10μg/ml）处理24 h，诱导炎症反应并研究分子信号、流量依赖性Ca 2+反应（3 ml/min ~ 10 ml/min）、ATP释放和ATP反应。治疗诱导了“rhEGC表型”，并导致分析的107个基因中58%的mRNA转录上调。调节基因包括炎症基因（54%/IP 10;IFNγ; CxCl 2; CCL 3; CCL 2;C3; s100 B; IL 1 β; IL 2R;TNFα; IL 4; IL 6; IL 8; IL 10; IL 12 A; IL 17 A; IL 22; IL 33），嘌呤基因（52%/P2 R2 A; P2 R2 B; P2 RY 1; P2 RY 2; P2 RY 6; P2 RX 3; P2 RX 7; AMPD 3; ENTPD 2; ENTPD 3; NADSYN 1），通道（40%/Panx 1; CHRNA 7; TRPV 1; TRPA 1）、囊泡转运蛋白（SYT 1、SYT 2、SNAP 25、SYP）、转录因子（relA/relB、SOCS 3、STAT 3、GATA_3、FOXP 3）、生长因子（IGFBP 5;GMCSF）、抗氧化剂基因（SOD 2; HMOX 1）和酶（NOS 2; TPH 2; CASP 3）（p<0.0001）。处理破坏了hEGC中的Ca 2+信号传导、ATP和机械/流动依赖性Ca 2+反应。ATP释放增加5倍，s100 B减少33%。rhEGC表型通过复杂的促炎途径级联来鉴定，其导致重要的分子和功能信号传导途径（Ca 2+、嘌呤能、机械感觉）的改变，这可能破坏GI运动。炎症诱导从ATP到ADP/腺苷/UTP信号传导的“嘌呤能开关”。研究结果对GI感染、IBD、POI、运动和GI疾病有影响。
Clinical observations or animal studies implicate enteric glial cells (EGC) in motility disorders, IBS, IBD, GI infections, post-operative ileus and slow transit constipation. Mechanisms underlying glial responses to inflammation in human GI tract are not understood. Our goal was to identify the ‘reactive human EGC phenotype’ induced by inflammation and probe its functional relevance. Human EGC in culture from 15 GI-surgical specimens were used to study gene expression, Ca2+ and purinergic signaling by Ca2+/fluo-4 imaging and mechanosensitivity. A nanostring-panel of 107 genes was designed as a read out of inflammation, transcription, purinergic signaling, vesicular transport-protein, channel, antioxidant and other pathways. A 24 h treatment with lipopolysaccharide (LPS, 200μg/ml) and interferon-γ (10μg/ml) was used to induce inflammation and study molecular signaling, flow-dependent Ca2+ responses from 3ml/min to 10ml/min, ATP release and ATP responses. Treatment induced a ‘rhEGC phenotype’ and caused upregulation in mRNA transcripts of 58% of 107 genes analyzed. Regulated genes included inflammatory genes (54%/IP10;IFNγ;CxCl2;CCL3;CCL2;C3;s100B;IL1β;IL2R;TNFα;IL4;IL6;IL8;IL10;IL12A;IL17A;IL22; IL33), purine-genes (52%/AdoR2A;AdoR2B;P2RY1;P2RY2;P2RY6;P2RX3;P2RX7;AMPD3;ENTPD2;ENTPD3; NADSYN1), channels (40%/Panx1;CHRNA7;TRPV1;TRPA1), vesicular-transporters (SYT1,SYT2,SNAP25,SYP), transcription factors (relA/relB,SOCS3,STAT3,GATA_3,FOXP3), growth factors (IGFBP5;GMCSF), antioxidant-genes (SOD2;HMOX1), and enzymes (NOS2;TPH2;CASP3)(p<0.0001). Treatment disrupted Ca2+ signaling, ATP and mechanical/flow-dependent Ca2+ responses in hEGC. ATP release increased 5-fold and s100B decreased 33%. The ‘rhEGC phenotype is identified by a complex cascade of pro-inflammatory pathways leading to alterations of important molecular and functional signaling pathways (Ca2+, purinergic, mechanosensory) that could disrupt GI motility. Inflammation induced a ‘purinergic switch’ from ATP to ADP/adenosine/UTP signaling. Findings have implications for GI infection, IBD, POI, motility and GI disorders.