Wip1 通过 ATM-p53 和 mTOR 信号通路影响角质细胞和内皮细胞的活性，从而调节伤口愈合。

Wip1 regulates wound healing by affecting activities of keratinocytes and endothelial cells through ATM-p53 and mTOR signaling.

机构信息

Department of Plastic Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

出版信息

Burns. 2023 Dec;49(8):1969-1982. doi: 10.1016/j.burns.2023.05.005. Epub 2023 May 12.

DOI:10.1016/j.burns.2023.05.005

PMID:37357059

Abstract

BACKGROUND

As a p53-regulated gene, Wip1 regulates proliferation, migration, apoptosis, and senescence of several type cells, but its biological functions in keratinocytes and endothelial cells which are involved wound healing are not fully understood. This study aims to reveal the function and underlying mechanism of Wip1 in wound healing using models of transgenic animal, keratinocytes, and endothelial cells.

METHODS

Using Wip1 knockout C57 BL/6 mice, we investigated effect of Wip1 deficiency on wound healing and angiogenesis; And using HaCaT and HUVEC as keratinocytes and endothelial cells, combined using primary keratinocytes from Wip1 knockout mice, we studied the effects of Wip1 knockdown/knockout or overexpression on proliferation, migration, and protein expressions of signaling components in ATM-p53 and mTOR pathway.

RESULTS

Wip1 deficiency in mice impaired the wound repair and endothelial angiogenesis, reduced the thickness of granulation tissue, and decreased the number of Ki67-positive cells and CD31 positive vessels in granulation tissue. Knockdown of Wip1 by shRNAs suppressed the proliferation and migration of HaCaT and HUVEC cells and induced notably apoptosis in the two cells. In western blot, Wip1 knockdown enriched p53 and ATM proteins, while decreased activated AKT, mTOR and activated S6 ribosomal protein (pS6) levels in HaCaT and HUVEC cells. Ectopic expression of Wip1 decreased the p53 and ATM proteins, while increased activated AKT, mTOR and pS6 levels in HaCaT and HUVEC cells. And in primary keratinocytes from mice tail skin, Wip1 knockout increased p53 and ATM, while decreased activated AKT, mTOR and pS6 protein levels.

CONCLUSION

Our study directly supports that Wip1 regulated skin wound healing possibly by affecting bioactivities including proliferation, migration and apoptosis of keratinocytes and endothelial cells at least through by modulating ATM-p53 and mTOR signaling.

摘要

背景

作为 p53 调控的基因，Wip1 调节多种类型细胞的增殖、迁移、凋亡和衰老，但它在参与伤口愈合的角质细胞和内皮细胞中的生物学功能尚未完全阐明。本研究旨在利用转基因动物、角质细胞和内皮细胞模型，揭示 Wip1 在伤口愈合中的功能和潜在机制。

方法

使用 Wip1 敲除 C57BL/6 小鼠，我们研究了 Wip1 缺乏对伤口愈合和血管生成的影响；并使用 HaCaT 和 HUVEC 作为角质细胞和内皮细胞，结合使用 Wip1 敲除小鼠的原代角质细胞，我们研究了 Wip1 敲低/敲除或过表达对增殖、迁移和 ATM-p53 和 mTOR 通路信号成分蛋白表达的影响。

结果

小鼠中 Wip1 的缺失损害了伤口修复和内皮血管生成，减少了肉芽组织的厚度，并减少了 Ki67 阳性细胞和 CD31 阳性血管在肉芽组织中的数量。shRNA 敲低 Wip1 抑制了 HaCaT 和 HUVEC 细胞的增殖和迁移，并显著诱导了这两种细胞的凋亡。在 Western blot 中，Wip1 敲低增加了 p53 和 ATM 蛋白的表达，同时降低了 HaCaT 和 HUVEC 细胞中激活的 AKT、mTOR 和激活的 S6 核糖体蛋白（pS6）的水平。Wip1 的过表达降低了 p53 和 ATM 蛋白的表达，同时增加了 HaCaT 和 HUVEC 细胞中激活的 AKT、mTOR 和 pS6 蛋白的水平。并且在来自小鼠尾皮的原代角质细胞中，Wip1 敲除增加了 p53 和 ATM 的表达，同时降低了激活的 AKT、mTOR 和 pS6 蛋白的水平。