Cellular Function of Annexin A1 Protein Mimetic Peptide Ac2-26 in Human Skin Keratinocytes HaCaT and Fibroblast Detroit 551 Cells.

Seong Min Kim,Preethi Vetrivel,Young Sil Kim,Hun Hwan Kim,Jung Eun Park,Sang Eun Ha,Gon Sup Kim,Jeong Doo Heo,Pritam Bhagwan Bhosale

doi:10.3390/nu12113261

Seong Min Kim, Preethi Vetrivel + Show 7 more

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https://doi.org/10.3390/nu12113261

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Abstract

Inflammation of the skin is the most common dermatological problem in human. The anti-inflammatory mediated responses of the skin cells provide a mechanism for combating these conditions. Annexin A1 (AnxA1) is one of the proteins that has been shown to have a potent anti-inflammatory effect. However, the effects and mechanisms of AnxA1 in skin keratinocyte and fibroblast have not been reported yet. In the current study, we hypothesized that Ac2-26, AnxA1 mimetic peptide, ameliorates inflammation and wrinkle formation in human skin cells. Therefore, we aimed to identify whether Ac2-26 has anti-inflammatory and anti-wrinkle effects in human keratinocyte (HaCaT) and fibroblast (Detroit 551) cells, respectively. Human HaCaT cells were stimulated by TNF-α/IFN-γ with or without Ac2-26, to identify the anti-inflammatory effect. Human Detroit 551 cells were treated with Ac2-26 to verify the anti-wrinkle effect. Initially, cell cytotoxicity was carried out in each cell line treated using Ac2-26 by MTT assay. Human MDA, IL-8, and procollagen secretion were detected by ELISA assay. The inflammatory chemokines were measured by qRT-PCR analysis. To demonstrate the mechanism, MAPK, NF-κB, JAK/STAT, and MMPs were analyzed by Western blotting. As a result, we identified that Ac2-26 significantly decreased the expression of TNF-α/IFN-γ-stimulated pro-inflammatory chemokines, including IL-1β, IL-6, IL-8, MDC, TARC, and TNF-α, by inhibiting the activation of MAPK, NF-κB, and JAK/STAT pathway in TNF-α/IFN-γ-stimulated HaCaT human keratinocytes. In addition, we also identified that Ac2-26 significantly induced collagen synthesis by generating pro-collagen, and suppressed collagen degradation by inhibiting the collagenase MMP-1 and MMP-8 expression. Collectively, these results suggest that Ac2-26 shows anti-inflammatory and anti-wrinkling effect. These effects may lead to the development of preventive and therapeutic application for inflammation-related skin disease and wrinkle formation.

Highlights

In human, the skin is the largest organ and protects from the outside environment by an epithelial barrier with abundant immune cells, such as lymphocytes and macrophages [1]
Th-2-associated chemokines have been recognized as a crucial mediator in chronic skin disease, suggesting that the regulation of TARC/CCL17 and MDC/CCL22 in keratinocytes may be used as an effective therapeutic target
The results show that Ac2-26 inhibited the production of chemokine MDC and IL-8 in a dosedose-dependent production of these chemokines, compared with the only tumor necrosis factor (TNF)-α- or IFN-γ-treated group

Summary

Introduction

The skin is the largest organ and protects from the outside environment by an epithelial barrier with abundant immune cells, such as lymphocytes and macrophages [1]. These immune cells are strongly associated with inflammatory skin responses to pathogens, but are involved in chronic inflammatory skin diseases such as atopic dermatitis and psoriasis [2]. Th-2-associated chemokines have been recognized as a crucial mediator in chronic skin disease, suggesting that the regulation of TARC/CCL17 and MDC/CCL22 in keratinocytes may be used as an effective therapeutic target

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