Peng Jiarui, Zhu Hong, Ruan Bin, Duan Zhisheng, Cao Mei
Department of Endocrinology, The Third People's Hospital of Yunnan Province, Kunming, China.
Department of Occupational Disease, The Third People's Hospital of Yunnan Province, Kunming, China.
J Diabetes Investig. 2025 Jan;16(1):60-71. doi: 10.1111/jdi.14327. Epub 2024 Nov 7.
Diabetic foot ulcers (DFU) are one of the most destructive complications of diabetes mellitus. The aim of this study was to link miR-155 and SOX2 with DFU to explore the regulation of wound healing by DFU and its potential mechanism.
Human keratinocytes (HaCaT) were induced with advanced glycation end products (AGEs) to construct DFU models in vitro. AGE-induced HaCaT cells were subjected to CCK-8 assays, flow cytometry, and wound healing assays to evaluate cell proliferation, apoptosis, and migration capacity, respectively. RT-qPCR and Western blotting were used to determine gene and protein expression levels, respectively. N6-methyladenosine (M6A) levels in total RNA were assessed using an M6A methylation quantification kit.
Our results suggested that the inhibition of miR-155 promoted wound healing in an in vitro DFU model, while the knockdown of HIF-1α reversed this process, and that HIF-1α was a target protein of miR-155. In addition, knockdown of HIF-1α promoted the m6A level of SOX2 mRNA, inhibited the expression of SOX2, and inhibited the activation of the EGFR/MEK/ERK signaling pathway, thus inhibiting the proliferation and migration of HaCaT cells and promoting the apoptosis of HaCaT cells, while overexpression of SOX2 reversed this effect. We also found that METTL3 knockdown had the opposite effect of HIF-1α knockdown.
Inhibition of miR-155 promoted the expression of HIF-1α and attenuated the m6A modification of SOX2 mRNA, thereby promoting the expression of SOX2 and activating the downstream EGFR/MEK/ERK signaling pathway to promote wound healing in an in vitro DFU model.
糖尿病足溃疡（DFU）是糖尿病最具破坏性的并发症之一。本研究旨在将miR-155和SOX2与DFU联系起来，探讨DFU对伤口愈合的调控及其潜在机制。
用人角质形成细胞（HaCaT）经晚期糖基化终末产物（AGEs）诱导，在体外构建DFU模型。对AGE诱导的HaCaT细胞分别进行CCK-8检测、流式细胞术和伤口愈合检测，以评估细胞增殖、凋亡和迁移能力。分别用RT-qPCR和蛋白质免疫印迹法测定基因和蛋白质表达水平。使用m6A甲基化定量试剂盒评估总RNA中的N6-甲基腺苷（M6A）水平。
我们的结果表明，在体外DFU模型中，抑制miR-155可促进伤口愈合，而敲低HIF-1α可逆转这一过程，且HIF-1α是miR-155的靶蛋白。此外，敲低HIF-1α可提高SOX2 mRNA的m6A水平，抑制SOX2的表达，并抑制EGFR/MEK/ERK信号通路的激活，从而抑制HaCaT细胞的增殖和迁移，促进HaCaT细胞的凋亡，而SOX2的过表达可逆转这一效应。我们还发现，敲低METTL3具有与敲低HIF-1α相反的作用。
在体外DFU模型中，抑制miR-155可促进HIF-1α的表达，减弱SOX2 mRNA的m6A修饰，从而促进SOX2的表达并激活下游EGFR/MEK/ERK信号通路，促进伤口愈合。
