miR-141-3p通过调控KEAP1/Nrf2轴抑制糖尿病创面的氧化应激损伤
MiR-141-3p Inhibits Oxidative Stress Damage in Diabetic Wounds by Regulating the KEAP1/Nrf2 Axis
刘洋 1张雪婷 2卢俊3
作者信息
- 1. 湖北医药学院附属太和医院骨科,湖北 十堰 442000
- 2. 湖北医药学院附属太和医院皮肤科,湖北 十堰 442000
- 3. 湖北医药学院附属太和医院麻醉科,湖北 十堰 442000
- 折叠
摘要
目的:探讨miR-141-3p对氧化应激状态下角质形成细胞的保护作用及其分子机制.方法:体外过氧化氢(H2O2)预处理角质细胞,选择 miR-141-3p mimics/inhibitor 以及 siKEAP1 技术研究 miR-141-3p 对于KEAP1/Nrf2 信号通路的影响.采用CCK-8 法检测细胞活力,qRT-PCR检测mRNA水平,Western-blot法检测蛋白表达水平,双荧光素酶报告基因系统检测miR-141-3p靶基因,DCFH-DA探针检测胞内ROS水平,酶联免疫吸附试验法检测抗氧化酶活性,EDU法检测细胞增殖功能,细胞划痕实验检测细胞迁移功能.结果:过表达miR-141-3p增强了H2O2 处理下角质形成细胞的增殖和迁移能力,降低了胞内ROS水平,并提高了SOD和GSH-px活性;miR-141 可靶向负调控KEAP1,上调Nrf2 和HO-1 表达,发挥抗氧化应激作用;siKEAP1 抑制了miR-141-3p促增殖、促迁移和抗氧化的作用.结论:miR-141 能够通过KEAP1/Nrf2 信号通路有效减轻H2O2 诱导的角质形成细胞氧化应激损伤,并促进其增殖和迁移,从而发挥促进糖尿病创面愈合的作用.
Abstract
Objective To explore the protective effect and molecular mechanism of miR-141-3p on keratinocytes under oxi-dative stress.Methods Keratinocytes were pretreated with hydrogen peroxide(H2O2)in vitro,and miR-141-3p mimics/inhibitor and siKEAP1 were employed to study the effect of miR-141-3p on the KEAP1/Nrf2 signaling pathway.Cell via-bility,mRNA expressions,protein expressions,miR-141-3p target genes,and intracellular ROS were detected using the CCK-8 assay,qRT-PCR,Western blot,dual-luciferase reporter assay system,and the DCFH-DA probe,respectively.Enzyme-linked immunosorbent assay,EDU method,and cell scratch assay were used to determine the antioxidant enzyme activity,cell proliferation function,and cell migration function,respectively.Results Overexpression of miR-141-3p en-hanced the proliferation and migration of keratinocytes under H2O2,reduced intracellular ROS,and increased SOD and GSH-px activities.MiR-141 could target and negatively regulate KEAP1,and up-regulate the expressions of Nrf2 and HO-1 to suppress oxidative stress.SiKEAP1 inhibited the pro-proliferation,pro-migration,and anti-oxidative effects of miR-141-3p.Conclusion miR-141 effectively reduces H2O2-induced oxidative stress damage in keratinocytes through the KEAP1/Nrf2 signaling pathway and promote their proliferation and migration,thereby facilitating diabetic wound healing.
关键词
糖尿病创面/创面愈合/氧化应激/miR-141-3p/Nrf2Key words
Diabetic wound/Wound healing/Oxidative stress/miR-141-3p/Nrf2引用本文复制引用
基金项目
国家自然科学基金(81772094)
国家自然科学基金(81974289)
出版年
2024
国家科技期刊平台
NSTL
万方数据