Abstract Egg white peptides are well known for their abundant species and biological properties. The aim of this study was to investigate the effect of egg white peptides on skin wound healing. We hydrolyzed the egg white powder using Alcalase and flavourzyme. Our transwell cell migration assay data showed that EWP2, which is one of the four fractions obtained from Sephadex G-15 isolation, promoted human skin fibroblast HSF cells migration. LC–MS/MS analysis results showed EWP2 were consist of 24 high abundant peptides. We randomly chose and synthesized four of these 24 peptides. Transwell assay data displayed that the egg white peptide KPHAEVVLR (KR-9) increased 8.2-fold of HSF cells migration at 200 μM compared with control. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sμlfophenyl)-2H-tetrazolium (MTS) assay results showed KR-9 did not lead to HSF cells over-proliferation. In vivo animal experiment data proved that KR-9 accelerated 50 % and 136 % mice skin wound healing rate at 50 μM and 100 μM. Furthermore, we explored the mechanism of KR-9. Our western blot and immunofluorescence results showed KR-9 enhanced the protein level of cell membrane heat shock protein 90α (Hsp90α). Thus, we conclude that egg white peptide KR-9 promotes skin fibroblasts migration and mice skin wound healing by stimulating cell membrane Hsp90α secretion.

中文翻译：

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蛋清肽 KPHAEVVLR 通过刺激细胞膜 Hsp90α 分泌促进皮肤成纤维细胞迁移和小鼠皮肤伤口愈合

摘要 蛋清肽以其丰富的物种和生物学特性而闻名。本研究的目的是研究蛋清肽对皮肤伤口愈合的影响。我们使用碱性蛋白酶和风味酶水解蛋清粉。我们的 transwell 细胞迁移测定数据显示，EWP2 是从 Sephadex G-15 分离中获得的四种组分之一，可促进人皮肤成纤维细胞 HSF 细胞迁移。LC-MS/MS 分析结果显示 EWP2 由 24 种高丰度肽组成。我们随机选择并合成了这 24 种肽中的四种。Transwell 测定数据显示，与对照相比，蛋清肽 KPHAEVVLR (KR-9) 在 200 μM 时使 HSF 细胞迁移增加了 8.2 倍。3-(4, 5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-sμlfo苯基)-2H-四唑鎓(MTS)测定结果显示KR-9没有导致HSF细胞过度增殖。体内动物实验数据证明，KR-9 在 50 μM 和 100 μM 下使小鼠皮肤伤口愈合速度加快了 50% 和 136%。此外，我们探索了 KR-9 的机制。我们的蛋白质印迹和免疫荧光结果显示 KR-9 增强了细胞膜热休克蛋白 90α (Hsp90α) 的蛋白质水平。因此，我们得出结论，蛋清肽 KR-9 通过刺激细胞膜 Hsp90α 分泌促进皮肤成纤维细胞迁移和小鼠皮肤伤口愈合。我们的蛋白质印迹和免疫荧光结果显示 KR-9 增强了细胞膜热休克蛋白 90α (Hsp90α) 的蛋白质水平。因此，我们得出结论，蛋清肽 KR-9 通过刺激细胞膜 Hsp90α 分泌促进皮肤成纤维细胞迁移和小鼠皮肤伤口愈合。我们的蛋白质印迹和免疫荧光结果显示 KR-9 增强了细胞膜热休克蛋白 90α (Hsp90α) 的蛋白质水平。因此，我们得出结论，蛋清肽 KR-9 通过刺激细胞膜 Hsp90α 分泌促进皮肤成纤维细胞迁移和小鼠皮肤伤口愈合。