反义lncRNA KRT-AS通过调控角蛋白基因表达促进糖尿病皮肤创面愈合

doi:10.3969/j.issn.1009-976X.2026.03.005

摘要/Abstract

摘要： 目的探讨反义长链非编码RNA（lncRNA）对角蛋白基因表达的调控和角质形成细胞功能的影响,并阐述其在糖尿病创面愈合中的作用。方法利用lncRNA芯片技术筛选糖尿病皮肤与正常皮肤差异表达的lncRNA,并通过cDNA末端快速扩增技术鉴定出一条由角蛋白基因反义链转录出的新型lncRNA KRT-AS。采用Western blot、实时荧光定量PCR、免疫荧光检测lncRNA KRT-AS对角蛋白14（Krt14）表达的调控作用。通过CCK-8、流式细胞分析、划痕实验评估其对角质形成细胞增殖、细胞周期和迁移能力的影响。构建糖尿病大鼠皮肤创面模型,观察局部过表达lncRNA KRT-AS对大鼠创面愈合的促进作用。通过核糖核酸酶保护实验和α-鹅膏蕈碱阻断实验,探讨lncRNA KRT-AS对Krt14 mRNA表达调控的机制。结果芯片分析结果提示,lncRNA KRT-AS在糖尿病皮肤中下调最为显著,下调倍数达68.34倍。角质形成细胞敲低lncRNA KRT-AS可抑制Krt14的mRNA和蛋白表达,同时抑制细胞增殖和迁移能力,并将细胞周期阻滞于G0/G1期。动物实验证实,局部过表达lncRNA KRT-AS可促进糖尿病创面愈合,增加表皮厚度,并上调Krt14的表达。机制研究发现,lncRNA KRT-AS可与Krt14 mRNA形成RNA-RNA双链结构,保护mRNA免受降解,从而提高Krt14 mRNA的表达水平。结论 LncRNA KRT-AS在糖尿病皮肤中表达下调。过表达lncRNA KRT-AS可通过增加Krt14 mRNA的稳定性,上调Krt14的表达,从而促进糖尿病创面愈合。LncRNA KRT-AS可作为糖尿病创面修复的潜在治疗靶点。

关键词: 糖尿病创面愈合, 反义长链非编码RNA, 角蛋白, 角质形成细胞, mRNA稳定性

Abstract: Objective This study aimed to investigate the effects of antisense long non-coding RNA （lncRNA） KRT-AS on keratin expression and keratinocyte function, and to elucidate its role in diabetic wound healing. Methods Differential lncRNA expression profiles between diabetic and non-diabetic skin were analyzed using microarray. A novel lncRNA, keratin antisense RNA （KRT-AS）, which is transcribed from the opposite strand of keratin genes, was identified and characterized by rapid amplification of cDNA ends. The effects of lncRNA KRT-AS on keratin 14 （Krt14） expression, proliferation, cell cycle, and migration were assessed in keratinocytes. The in vivo effect of lncRNA KRT-AS was evaluated using a diabetic rat wound healing model. RNA stability was assessed using ribonuclease protection assays and α-amanitin treatment. Results LncRNA KRT-AS was the most significantly downregulated lncRNA in diabetic skin. Knockdown of lncRNA KRT-AS in keratinocytes decreased cell proliferation, induced G0/G1 phase cell cycle arrest, inhibited cell migration, and suppressed Krt14 expression. In vivo experiments suggested lncRNA KRT-AS accelerated diabetic wound closure, increased epidermal thickness and upregulated Krt14 expression in diabetic skin. Mechanistically, lncRNA KRT-AS formed an RNA-RNA duplex with Krt14 mRNA, protecting the mRNA from degradation, thereby enhancing the expression level of Krt14 mRNA. Conclusion LncRNA KRT-AS is downregulated in diabetic skin. Overexpression of lncRNA KRT-AS promotes diabetic wound healing by enhancing the stability of Krt14 mRNA in keratinocytes. LncRNA KRT-AS represents a potential therapeutic target for diabetic wound repair.

Key words: diabetic wound healing, antisense lncRNA, keratin, keratinocyte, mRNA stability

中图分类号:

R587.2
R641

韩雨婷, 杨川, 胡梦蝶. 反义lncRNA KRT-AS通过调控角蛋白基因表达促进糖尿病皮肤创面愈合[J]. 岭南现代临床外科, 2026, 26(03): 167-175.

HAN Yuting, YANG Chuan, HU Mengdie. Antisense lncRNA KRT-AS promotes diabetic cutaneous wound healing by regulating keratin expression[J]. Lingnan Modern Clinics In Surgery, 2026, 26(03): 167-175.

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