Effects of the neuropeptides substance P, calcitonin gene-related peptide, vasoactive intestinal polypeptide and galanin on the production of nerve growth factor and inflammatory cytokines in cultured human keratinocytes
Effects of the neuropeptides substance P, calcitonin gene-related peptide, vasoactive intestinal polypeptide and galanin on the production of nerve growth factor and inflammatory cytokines in cultured human keratinocytes
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DOI：
10.1016/j.npep.2006.06.002
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发表时间：
2006-08-01
期刊：
影响因子：
2.9
通讯作者：
Husz, Sandor
中科院分区：
文献类型：
--
作者：
Dallos, Attila;Kiss, Maria;Husz, Sandor
从皮肤感觉神经末梢释放的神经肽具有神经递质和免疫调节作用；它们发挥促有丝分裂作用，可以影响皮肤中不同类型细胞的功能。本研究旨在系统研究神经肽P物质(SP)、降钙素基因相关肽(CGRP)、血管活性肠肽(VIP)和甘丙素(GAL)对角质形成细胞产生炎性细胞因子(IL-1α、IL-8和TNF-α)的影响，并探讨它们在神经生长因子(NGF)及其前体分子(ProNGF)产生和分泌中的作用。用10(-8)M SP、CGRP、VIP或GAL处理培养的正常人角质形成细胞30min。在不同的时间间隔后，收获细胞进行总RNA提取；此外，收集细胞裂解产物和培养上清。用Q-RT-PCR法检测神经肽对不同细胞因子和神经生长因子基因表达的影响，用ELISA法和Western blotting法检测神经肽的蛋白表达水平。四种神经肽均可诱导IL-1α、IL-8和肿瘤坏死因子-α的表达增加。神经肽处理的细胞培养上清液中IL-1α蛋白含量增加，IL-8分泌轻度增加，而未检测到肿瘤坏死因子-α的分泌。四种神经肽均不同程度地促进了NGF基因的表达。在角质形成细胞的细胞裂解产物中，只能检测到proNGF，其在神经肽处理的细胞中的浓度大约是时间匹配对照组的两倍。对照培养物和神经肽处理的细胞培养物都能分泌ProNGF和成熟的NGF，但神经肽处理的细胞培养物产生的NGF样免疫反应物的数量显著增加(3-7倍)。结果表明，损伤刺激后皮肤神经释放的神经肽能够诱导IL-1α和IL-8的产生上调，并能影响角质形成细胞ProNGF/NGF的表达和分泌。这些发现可能有助于理解神经对皮肤健康和疾病的影响。(C)2006爱思唯尔有限公司。保留所有权利。
Neuropeptides released from the cutaneous sensory nerve endings have neurotransmitter and immunoregulatory roles; they exert mitogenic actions and can influence the functions of different cell types in the skin. The aims of this study were a systematic investigation of the effects of the neuropeptides substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and galanin (GAL) on the inflammatory cytokine production (IL-1 alpha, IL-8 and TNF-alpha) of the keratinocytes, and a study of their role in the production and secretion of nerve growth factor (NGF) and its precursor molecule (proNGF). Cultures of normal human keratinocytes were treated with 10(-8) M SP, CGRP, VIP or GAL for 30 min. After different time intervals, cells were harvested for total RNA isolation; in addition, cell lysates and supernatants were collected. The effects of the neuropeptides on the mRNA expressions of the different cytokines and NGF were investigated by Q-RT-PCR and the protein levels were studied by means of ELISA assays and Western blotting. Each of the four neuropeptides induced increases in the expressions of IL-1 alpha, IL8 and TNF-a mRNA. Increases appeared in the amount of the IL-1 alpha protein in the supernatants of neuropeptide-treated cells, and the IL-8 secretion was mildly elevated, while secretion of TNF-a remained undetectable. The four neuropeptides increased the NGF mRNA expression to different extents. In the cell lysates of the keratinocytes, only proNGF could be detected, its concentration in the neuropeptide-treated cells being approximately twice that in the time-matched controls. Both control cultures and neuropeptide-treated cultures were found to secrete proNGF and mature NGF, but neuropeptide-treated cell cultures produced markedly higher (3-7-fold) amounts of NGF-like immunoreactive materials. The results demonstrated that neuropeptides released from cutaneous nerves after an injurious stimulus are able to induce an upregulation of IL-1 alpha and IL-8 production; they are additionally able to influence the expressions of proNGF/NGF and their secretion from the keratinocytes. These findings may contribute toward an understanding of the neural influence on skin health and disease. (c) 2006 Elsevier Ltd. All rights reserved.