Human catestatin enhances migration and proliferation of normal human epidermal keratinocytes
Human catestatin enhances migration and proliferation of normal human epidermal keratinocytes
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DOI：
10.1016/j.jdermsci.2011.08.001
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发表时间：
2011-11-01
影响因子：
4.6
通讯作者：
Ogawa, Hideoki
中科院分区：
文献类型：
--
作者：
Hoq, Md Imranul;Niyonsaba, Francois;Ogawa, Hideoki
背景资料：皮肤来源的抗微生物肽，如人β-防御素和cathelicidins，通过灭活微生物积极促进宿主防御。Catestatin是一种影响人类自主神经功能的神经内分泌肽，最近在损伤/感染后的角质形成细胞中检测到，它抑制病原体的生长。目的：研究人catestatin及其变异体对角质形成细胞迁移和增殖的影响，并探讨其可能的信号转导机制。通过BrdU掺入、细胞计数和细胞周期分析来评价细胞增殖。使用荧光钙测定试剂盒测量细胞内Ca 2+动员。表皮生长因子受体（EGFR），Akt，和MAPKs的磷酸化通过Western blotting.Results：Catestatin及其变体剂量依赖性地增强角质形成细胞的迁移和增殖。此外，catestatin肽增加细胞内Ca 2+动员，并诱导表皮生长因子受体，Akt，细胞外信号调节激酶（ERK），和p38在角质形成细胞的磷酸化。通过百日咳毒素的特异性抑制作用证明，catestatin肽诱导角质形成细胞迁移和增殖涉及G蛋白、磷脂酶C、EGFR、PI 3-激酶、ERK和p38（G蛋白抑制剂），U-73122（磷脂酶C抑制剂），AG 1478（EGFR抑制剂）、抗EGFR抗体、渥曼青霉素（PI 3-激酶抑制剂），U 0126（ERK抑制剂）和SB 203580（p38抑制剂）。除了抑制皮肤病原体的生长外，catestatin肽还可以通过增强伤口部位的角质细胞迁移和增殖而有助于皮肤伤口闭合。(C)2011年日本皮肤病研究学会。由Elsevier爱尔兰有限公司出版。保留所有权利。
Background: Skin-derived antimicrobial peptides, such as human beta-defensins and cathelicidins, actively contribute to host defense by inactivating microorganisms. Catestatin, a neuroendocrine peptide that affects human autonomic functions, has recently been detected in keratinocytes upon injury/infection where it inhibits the growth of pathogens. Human catestatin exhibits three single nucleotide polymorphisms: Gly364Ser, Pro370Leu, and Arg374Gln.Objective: To investigate the effects of human catestatin and its variants on keratinocyte migration and proliferation, and to elucidate the possible signaling mechanisms involved.Methods: The migration of normal human keratinocytes was analyzed using Boyden microchamber assay and in vitro wound closure assay. Cell proliferation was evaluated by BrdU incorporation, cell count assay and cell cycle analysis. Intracellular Ca2+ mobilization was measured using a fluorescent calcium assay kit. The phosphorylation of epidermal growth factor receptor (EGFR), Akt, and MAPKs was determined by Western blotting.Results: Catestatin and its variants dose-dependently enhanced keratinocyte migration and proliferation. Moreover, catestatin peptides increased intracellular Ca2+ mobilization and induced the phosphorylation of EGFR, Akt, extracellular signal-regulated kinase (ERK), and p38 in keratinocytes. The induction of keratinocyte migration and proliferation by catestatin peptides involved G-proteins, phospholipase C, EGFR, PI3-kinase, ERK, and p38, as evidenced by the specific inhibitory effects of pertussis toxin (G-protein inhibitor), U-73122 (phospholipase C inhibitor), AG1478 (EGFR inhibitor), anti-EGFR antibody, wortmannin (PI3-kinase inhibitor), U0126 (ERK inhibitor), and SB203580 (p38 inhibitor), respectively.Conclusion: Besides inhibiting the growth of skin pathogens, catestatin peptides may also contribute to cutaneous wound closure by enhancing keratinocyte migration and proliferation at the wound site. (C) 2011 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.