N6-甲基腺苷甲基转移酶威尔姆斯瘤1相关蛋白通过表观遗传激活DNA甲基转移酶1来阻碍糖尿病伤口愈合。

N6-methyladenosine methyltransferase Wilms tumor 1-associated protein impedes diabetic wound healing through epigenetically activating DNA methyltransferase 1.

作者信息

Xiao Ren-Jie, Wang Tian-Jiao, Wu Dan-Yin, Yang Shui-Fa, Gao Hai, Gan Pei-Dong, Yi Yang-Yan, Zhang You-Lai

机构信息

Department of Anesthesiology, The Second Affiliated Hospital, Jiangxi Medical College of Nanchang University, Nanchang 330006, Jiangxi Province, China.

State Key Laboratory of Advanced Medical Materials and Devices, Tianjin Key Laboratory of Biomedical Materials, Key Laboratory of Biomaterials and Nanotechnology for Cancer Immunotherapy, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300192, China.

出版信息

World J Diabetes. 2025 Mar 15;16(3):102126. doi: 10.4239/wjd.v16.i3.102126.

DOI:10.4239/wjd.v16.i3.102126

PMID:40093271

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11885966/

Abstract

BACKGROUND

Diabetic wound injury is a significant and common complication in individuals with diabetes. N6-methyladenosine (m6A)-related epigenetic regulation is widely involved in the pathogenesis of diabetes complications. However, the function of m6A methyltransferase Wilms tumor 1-associated protein (WTAP) in diabetic wound healing remains elusive.

AIM

To investigate the potential epigenetic regulatory mechanism of WTAP during diabetic wound healing.

METHODS

Human umbilical vein endothelial cells (HUVECs) were induced with high glucose (HG) to establish cell model. Male BALB/c mice were intraperitoneally injected with streptozotocin to mimic diabetes, and full-thickness excision was made to mimic diabetic wound healing. HG-induced HUVECs and mouse models were treated with WTAP siRNAs and DNA methyltransferase 1 (DNMT1) overexpression vectors. Cell viability and migration ability were detected by cell counting kit-8 and Transwell assays. angiogenesis was measured using a tube formation experiment. The images of wounds were captured, and re-epithelialization and collagen deposition of skin tissues were analyzed using hematoxylin and eosin staining and Masson's trichrome staining.

RESULTS

The expression of several m6A methyltransferases, including METTL3, METTL14, METTL16, KIAA1429, WTAP, and RBM15, were measured. WTAP exhibited the most significant elevation in HG-induced HUVECs compared with the normal control. WTAP depletion notably restored cell viability and enhanced tube formation ability and migration of HUVECs suppressed by HG. The unclosed wound area of mice was smaller in WTAP knockdown-treated mice than in control mice at nine days post-wounding, along with enhanced re-epithelialization rate and collagen deposition. The m6A levels on DNMT1 mRNA in HUVECs were repressed by WTAP knockdown in HUVECs. The mRNA levels and expression of DNMT1 were inhibited by WTAP depletion in HUVECs. Overexpression of DNMT1 in HUVECs notably reversed the effects of WTAP depletion on HG-induced HUVECs.

CONCLUSION

WTAP expression is elevated in HG-induced HUVECs and epigenetically regulates the m6A modification of DNMT1 to impair diabetic wound healing.

摘要

背景

糖尿病伤口损伤是糖尿病患者中一种严重且常见的并发症。N6-甲基腺苷（m6A）相关的表观遗传调控广泛参与糖尿病并发症的发病机制。然而，m6A甲基转移酶威尔姆斯瘤1相关蛋白（WTAP）在糖尿病伤口愈合中的作用仍不清楚。

目的

探讨WTAP在糖尿病伤口愈合过程中的潜在表观遗传调控机制。

方法

用高糖（HG）诱导人脐静脉内皮细胞（HUVECs）建立细胞模型。雄性BALB/c小鼠腹腔注射链脲佐菌素以模拟糖尿病，并进行全层切除以模拟糖尿病伤口愈合。用WTAP小干扰RNA（siRNAs）和DNA甲基转移酶1（DNMT1）过表达载体处理HG诱导的HUVECs和小鼠模型。通过细胞计数试剂盒-8和Transwell实验检测细胞活力和迁移能力。使用管腔形成实验测量血管生成。拍摄伤口图像，并用苏木精-伊红染色和Masson三色染色分析皮肤组织的再上皮化和胶原沉积。

结果

检测了几种m6A甲基转移酶的表达，包括METTL3、METTL14、METTL16、KIAA1429、WTAP和RBM15。与正常对照相比，WTAP在HG诱导的HUVECs中升高最为显著。WTAP缺失显著恢复了细胞活力，并增强了HG抑制的HUVECs的管腔形成能力和迁移能力。在伤口处理9天后，WTAP敲低处理的小鼠的未闭合伤口面积比对照小鼠小，同时再上皮化率和胶原沉积增加。HUVECs中WTAP敲低可抑制HUVECs中DNMT1 mRNA上的m6A水平。HUVECs中WTAP缺失可抑制DNMT1的mRNA水平和表达。HUVECs中DNMT1的过表达显著逆转了WTAP缺失对HG诱导的HUVECs的影响。