Substance P induced preprotachykinin-A mRNA, neutral endopeptidase mRNA and substance P in cultured normal fibroblasts
Substance P induced preprotachykinin-A mRNA, neutral endopeptidase mRNA and substance P in cultured normal fibroblasts
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DOI：
10.1159/000057749
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发表时间：
2002-04-01
影响因子：
2.8
通讯作者：
Katayama, I
中科院分区：
文献类型：
--
作者：
Bae, SJ;Matsunaga, Y;Katayama, I
在某些皮肤疾病中，压力可以调节皮肤表现的诱导和/或进展。然而，对神经内分泌和皮肤免疫系统中的回路知之甚少。为了解决这个问题，我们分析了由培养的正常人成纤维细胞组成的皮肤的主要人口，并可能增加压力诱导的皮肤炎症反应的P物质（SP）的自分泌诱导的调节机制。在非刺激条件下，正常成纤维细胞表达适量的前速激肽-A（PPT-A），SP mRNA的前体，和外源性SP显着上调PPT-A mRNA的表达。SP刺激后1-3 h，成纤维细胞内SP肽和SP mRNA的表达达到最大值，而SP刺激24 h后，成纤维细胞内具有SP水解活性的细胞表面肽中性内肽酶（NEP）的表达增加。向成纤维细胞施用NEP抑制剂（磷胺）可诱导更高的SP产生。此外，神经激肽（NK）受体拮抗剂（Spantide，FK 224和FK 888）和蛋白质合成抑制剂（放线菌酮）抑制SP的产生的30-40%的控制响应。免疫组化结果显示，SP刺激成纤维细胞后，SP在细胞质内呈特异性染色。最后，我们证实成纤维细胞中表达的PPT-A的核苷酸序列与人PPT-AcDNA完全一致。本文首次报道正常人皮肤成纤维细胞对外源性SP可诱导SP mRNA、NEP mRNA和SP肽的表达，并初步证实成纤维细胞源性SP可能在某些皮肤疾病的诱导和加速中起重要作用。版权所有（C）2002 S. Karger AG，巴塞尔。
In certain skin diseases, stress can modulate the induction and/or progression of cutaneous manifestations. However, little is known about the circuit in neuroendocrine and in the immune systems of the skin. To address this question, we have analyzed the regulatory mechanisms of autocrine induction of substance P (SP) by cultured normal human fibroblasts that compose the major population of the skin and might augment stress-induced skin inflammatory responses. In nonstimulated conditions, normal fibroblasts express a moderate amount of preprotachykinin-A (PPT-A), a precursor of SP mRNA, and exogenous SP significantly upregulated PPT-A mRNA expression. Maximum response of SP peptide and SP mRNA in fibroblasts was observed 1-3 h after stimulation with SP. In contrast, the expression of neutral endopeptidase (NEP), a cell surface peptide with hydrolyzing activity of SP, was increased in fibroblasts stimulated with SP after 24 h. The administration of NEP inhibitor (phosphoramidon) to the fibroblasts induced higher SP production. In addition, the neurokinin (NK) receptor antagonists (spantide, FK224 and FK888) and protein synthesis inhibitor (cycloheximide) inhibited SP production by 30-40% of control response. In immuno-staining study, specific cytoplasmic staining of SP was observed in fibroblasts stimulated with SP. Finally, we confirmed that the nucleotide sequence of the PPT-A expressed in fibroblasts perfectly corresponded to the gene bank human PPT-A cDNA. This is the first report that SP mRNA, NEP mRNA and SP peptide can be induced by normal human skin fibroblasts in response to exogenous SP, and that fibroblast-derived SP might play an important role in the induction and acceleration of certain cutaneous diseases. Copyright (C) 2002 S. Karger AG, Basel.