Angiogenic peptide (AG)-30/5C activates human keratinocytes to produce cytokines/chemokines and to migrate and proliferate via MrgX receptors

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• 作者：Chanisa Kiatsurayanon^a ; ^b ; ^c ; Franç ; ois Niyonsaba^a ; ^d ; ^{francois@juntendo.ac.jp" class="auth_mail" title="E-mail the corresponding author} ; Panjit Chieosilapatham^a ; ^b ; Ko Okumura^a ; Shigaku Ikeda^a ; ^b ; Hideoki Ogawa^a
• 关键词：AG-30 ; angiogenic peptide-30 ; hBD ; human β-defensin ; HDP ; host defense peptide ; IL ; interleukin ; LPS ; lipopolysaccharide ; MrgX ; Mass-related G-protein coupled receptor X ; PGN ; peptidoglycan ; poly I ; C ; polyinosinic ; polycytidylic acid ; TGF ; transforming growth factor ; TLR ; toll-like receptor
• 刊名：Journal of Dermatological Science
• 出版年：2016
• 出版时间：September 2016
• 年：2016
• 卷：83
• 期：3
• 页码：190-199
• 全文大小：2128 K

文摘

In addition to their antimicrobial activities, antimicrobial peptides, also known as host defense peptides (HDPs) activate keratinocytes; promote wound healing; and improve the skin barrier. AG-30/5C is a novel angiogenic HDP that activates various functions of fibroblasts and endothelial cells, including cytokine/chemokine production and wound healing.

Objectives

To investigate whether AG-30/5C activates human keratinocytes and to examine the underlying mechanisms.

Methods

Production of cytokines/chemokines was assessed by ELISA. Expression of Mas-related G-protein coupled receptors X (MrgXs) in keratinocytes was determined by real-time PCR and Western blot. MAPK and NF-κB activation was analysed by Western blot. Cell migration was assessed by chemotaxis microchamber and in vitro wound closure assay, whereas cell proliferation was analysed using an XTT assay.

Results

We found that AG-30/5C was more efficient than its parent peptide AG-30 in increasing the production of various cytokines/chemokines and promoting keratinocyte migration and proliferation. Furthermore, MrgX3 and MrgX4 receptors were constitutively expressed in keratinocytes at higher levels than MrgX1 and MrgX2, and were up-regulated upon stimulation with TLR ligands. Because MrgX3 and MrgX4 siRNAs suppressed AG-30/5C-mediated cytokine/chemokine production, keratinocyte migration and proliferation, we propose that AG-30/5C utilizes these MrgXs to stimulate keratinocytes. In addition, AG-30/5C-induced activation of keratinocytes was controlled by MAPK and NF-κB pathways, as evidenced by the inhibitory effects of ERK-, JNK-, p38- and NF-κB-specific inhibitors. Indeed, we confirmed that AG-30/5C enhanced phosphorylation of MAPKs and IκB.

Conclusions

Our findings provide novel evidence that AG-30/5C may be a useful therapeutic agent for wound healing by activating human keratinocytes.