MicroRNA-221-3p Targets THBS1 to Promote Wound Healing in Diabetes.
MicroRNA-221-3p Targets THBS1 to Promote Wound Healing in Diabetes.
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DOI：
10.2147/dmso.s424847
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Zhang Q
中科院分区：
文献类型：
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作者：
Hu K;Liu X;Chang H;Zhang Y;Zhou H;Liu L;Zhang X;Jiao Z;Shen B;Zhang Q
糖尿病足溃疡（DFU）是糖尿病的一种严重并发症，以慢性足溃疡、伤口愈合不良和持续性炎症为特征。MiR-221- 3 p作为microRNA，在以前的研究中已被证明可以加速WH，但其潜在机制尚不清楚。在这项研究中，我们的目的是确定miR-221- 3 p如何通过靶向THBS 1影响WH。表皮注射miRNA-221 - 3 p阿戈米尔对糖尿病创面愈合的影响体外产生的人永生化角质形成细胞（HaCaT细胞）用miR-模拟物转染，阴性对照用高葡萄糖处理。分别采用细胞凋亡实验和微管形成实验观察miRNA-221- 3 p对细胞凋亡和血管生成的影响。通过荧光素酶报告基因分析检测miR-221- 3 p与THBS 1的3 'UTR中预测的靶位点的直接靶相互作用。培育miRNA-221敲除小鼠用于实验验证。我们发现正常小鼠和糖尿病小鼠伤口边缘的miRNA-221- 3 p过表达可以促进WH。相比之下，miR-221敲除小鼠的WH延迟，增加上皮细胞凋亡和减少真皮中的血管生成。发现miR-221- 3 p抑制HaCaT细胞中的凋亡，并增强共培养的人脐静脉内皮细胞（HUVECs）中的血管生成。生物信息学分析以及双荧光素酶报告基因分析显示miR-221- 3 p靶向THBS 1的3 bp非翻译区。我们的研究结果表明，miR-221- 3 p可能通过抑制THBS 1对糖尿病WH产生重要影响，miR-221- 3 p的缺乏可能导致2型糖尿病慢性伤口愈合受损。因此，未来开发药物，如miR-221- 3 p的化学修饰类似物，可以使DFU患者受益。
Diabetes foot ulcer (DFU) is a serious complication of diabetes characterized with chronic foot ulceration, poor wound healing (WH), and persistent inflammation. MiR-221-3p, as microRNA, has been shown to accelerate WH in previous study, but the underlying mechanisms are poorly understood. In this study, we aimed to determine how miR-221-3p influences WH by targeting THBS1. The effect of miRNA-221-3p on wound healing of diabetes by epidermal injection of miRNA-221-3p agomir. In vitro generated human immortalized keratinocytes (HaCaT cells) were transfected with miR-mimics and negative control with high glucose treatment. The effects of miRNA-221-3p on cell apoptosis and angiogenesis using cell apoptosis assay and the tube formation assay, respectively. Direct target interaction of miR-221-3p and predicted target sites in 3’UTR of THBS1 were examined by luciferase reporter gene assay. Breeding miRNA-221 knockout mice for experimental verification. We found that miRNA-221-3p overexpression at the wound edge of normal mice and diabetes mice can promote WH. As contrast, WH of miR-221 knockout mice delayed with increased epithelial apoptosis and reduced angiogenesis in the dermis. miR-221-3p was found to inhibit apoptosis in HaCaT cells, and enhanced angiogenesis in human umbilical vein endothelial cells (HUVECs) that were co-cultured. Bioinformatics analysis as well as the dual-luciferase reporter assay revealed miR-221-3p to target 3ʹ untranslated region of THBS1. Our findings suggested miR-221-3p might exert an essential impact on diabetes WH via inhibition of THBS1 and lack of miR-221-3p possibly results in impaired healing in chronic wounds of type 2 diabetes. Therefore, developing medicine such as chemically modified analogs of miR-221-3p in future could benefit patients with DFU.