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首页> 外文期刊>International journal of colorectal disease. >Sensory neuropeptides and epithelial cell restitution: the relevance of SP- and CGRP-stimulated mast cells.
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Sensory neuropeptides and epithelial cell restitution: the relevance of SP- and CGRP-stimulated mast cells.

机译：感觉神经肽和上皮细胞恢复：SP和CGRP刺激的肥大细胞的相关性。

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BACKGROUND: Calcitonin-gene-related peptide (CGRP) and substance P (SP) are neurotransmitters of extrinsic primary afferent neurons located within the dorsal root ganglia. In experimental models of colitis in rats and rabbits, a protective role of SP and CGRP on intestinal mucosa was presumed. The mucosal protection partly depends on a CGRP-mediated modulation of mucosal blood flow. Limited data are available regarding CGRP- or SP-mediated effects on epithelial cell restitution. Having shown earlier that SP-stimulated fibroblasts but not CGRP-stimulated fibroblasts induce epithelial cell migration in vitro, the aim of this study was to explore whether mast cells mediate effects of SP and CGRP on epithelial cell restitution in vitro. METHODS: Mast cells (C57) were exposed to SP [10(-12)-10(-6 M)] and CGRP [10(-12)-10(-7 M)]. After a 24-h incubation period, the cell supernatants (conditioned media, CDM) were taken from mast cell cultures and directly applied to rat intestinal epithelial cell lines-18 or Caco-2 monolayers, which had been wounded with a razor blade 24 h prior to the experiments. Epithelial cell migration was assessed by counting cells across the wound edge and epithelial cell proliferation was measured using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium bromide test. RESULTS: CGRP significantly induced epithelial cell migration and proliferation via mast cells when supernatants were directly applied to epithelial cells in vitro. The effects on epithelial cell migration were abolished after neutralizing anti-transforming growth factor-beta (TGF-beta) had been added to the cell cultures. SP had no effects on epithelial cells following stimulation of mast cells. CONCLUSION: CGRP modulates epithelial cell restitution in vitro mediated by mast cells. The CGRP- and mast-cell-induced epithelial cell migration is TGF-beta dependent. This observation underlines an important role for extrinsic primary afferent neurons in mucosal defence and repair and in keeping the mucosal homeostasis. This knowledge leads to a better understanding of the interaction of the enteric nervous system and wound healing and may, in the future, lead to new therapeutic approaches to inflammatory diseases of the intestine.
机译：背景：降钙素基因相关肽（CGRP）和P物质（SP）是位于背根神经节内的外在初级传入神经元的神经递质。在大鼠和兔子的结肠炎实验模型中，推测SP和CGRP对肠粘膜有保护作用。粘膜保护部分取决于CGRP介导的对粘膜血流的调节。关于CGRP或SP介导的对上皮细胞恢复的影响的数据有限。早先已证明SP刺激的成纤维细胞而不是CGRP刺激的成纤维细胞在体外诱导上皮细胞迁移，本研究的目的是探讨肥大细胞是否介导SP和CGRP对体外上皮细胞恢复的影响。方法：将肥大细胞（C57）暴露于SP [10（-12）-10（-6 M）]和CGRP [10（-12）-10（-7 M）]。孵育24小时后，从肥大细胞培养物中提取细胞上清液（条件培养基，CDM），并直接应用于已用剃须刀受伤的大鼠肠道上皮细胞系18或Caco-2单层。实验之前。通过计数伤口边缘的细胞来评估上皮细胞的迁移，并使用3- [4，5-二甲基噻唑-2-基] -2，5-二苯基-四唑溴化物测试来测量上皮细胞的增殖。结果：当上清液直接应用于体外上皮细胞时，CGRP通过肥大细胞显着诱导上皮细胞迁移和增殖。在将中和的抗转化生长因子-β（TGF-β）加入细胞培养物中后，对上皮细胞迁移的影响被消除。肥大细胞刺激后，SP对上皮细胞无影响。结论：CGRP在肥大细胞介导的体外调节上皮细胞的重建。 CGRP和肥大细胞诱导的上皮细胞迁移是TGF-β依赖性的。该观察结果强调了外源性初级传入神经元在粘膜防御和修复以及保持粘膜稳态方面的重要作用。这些知识可以使人们更好地了解肠神经系统与伤口愈合之间的相互作用，并且在将来可能会导致针对肠炎性疾病的新治疗方法。

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