Human ENS regulates the intestinal epithelial barrier permeability and a tight junction-associated protein ZO-1 via VIPergic pathways
Human ENS regulates the intestinal epithelial barrier permeability and a tight junction-associated protein ZO-1 via VIPergic pathways
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DOI：
10.1152/ajpgi.00066.2003
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发表时间：
2003-11-01
影响因子：
4.5
通讯作者：
Jarry, A
中科院分区：
文献类型：
--
作者：
Neunlist, M;Toumi, F;Jarry, A
虽然肠神经系统（ENS）已被证明调节多种粘膜功能，但其在人体肠上皮屏障的生理控制中的作用尚不清楚。本研究的目的是研究ENS是否能够调节上皮屏障通透性和一个关键的紧密连接相关蛋白，闭塞带蛋白-1 （ZO-1）。因此，我们建立了一个共培养模型，由包含粘膜下层神经网络的人粘膜下层和人极化结肠上皮单层（HT29-Cl）组成。16E或Caco-2)。电场刺激（EFS）激活粘膜下神经元。通过测量细胞旁通透性标志物（fitc -葡聚糖或fitc -菊糖）穿过上皮单层的通量来评估通透性。采用免疫荧光、定量免疫印迹和实时RT-PCR检测ZO-1的表达。通过共培养模型，我们发现粘膜下神经元的EFS导致fitc -葡聚糖或fitc -菊糖通量减少，这被TTX阻断。在HT29-Cl。16E，粘膜下神经元的激活作用被VIP受体拮抗剂（VIPra）阻断，并由VIP复制。此外，在HT29-Cl中评估ZO-1的表达（mRNA，蛋白）。粘膜下神经元激活后，16E显著升高。这些对ZO-1表达的影响被TTX和VIPra阻断，并被VIP复制。综上所述，我们的研究结果强烈提示了VIPergic粘膜下神经元通路对肠上皮屏障通透性和ZO-1表达的调节作用。
Although the enteric nervous system ( ENS) has been shown to regulate various mucosal functions, its role in the physiological control of the human intestinal epithelial barrier is unknown. The aim of this study was to investigate whether the ENS is able to modulate epithelial barrier permeability and a key tight junction-associated protein, zonula occludens-1 (ZO-1). Therefore, we developed a coculture model, consisting of human submucosa containing the submucosal neuronal network and human polarized colonic epithelial monolayers (HT29-Cl.16E or Caco-2). Submucosal neurons were activated by electrical field stimulation (EFS). Permeability was assessed by measuring the flux of paracellular permeability markers ( FITC-dextran or FITC-inulin) across epithelial monolayers. Expression of ZO-1 was determined by immunofluorescence, quantitative immunoblot analysis, and real time RT-PCR. Using the coculture model, we showed that EFS of submucosal neurons resulted in a reduction in FITC-dextran or FITC-inulin fluxes, which was blocked by TTX. In HT29-Cl.16E, the effect of submucosal neuron activation was blocked by a VIP receptor antagonist (VIPra) and reproduced by VIP. Furthermore, ZO-1 expression ( mRNA, protein) assessed in HT29-Cl.16E, was significantly increased after submucosal neuron activation by EFS. These effects on ZO-1 expression were blocked by TTX and VIPra and reproduced by VIP. In conclusion, our results strongly suggest a modulatory role of VIPergic submucosal neuronal pathways on intestinal epithelial barrier permeability and ZO-1 expression.