Li Weifang, Xu Gengchen, Chai Gregory W, Ball Alexander, Zhang Qiuwang, Kutryk Michael J B
Department of Geriatric Endocrinology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Division of Cardiology, Keenan Research Center for Biomedical Science, St. Michael's Hospital, Unity Health Toronto, University of Toronto, Toronto, ON, Canada.
Sci Rep. 2025 Feb 25;15(1):6738. doi: 10.1038/s41598-025-91100-1.
MicroRNAs, a class of small non-coding RNA molecules that regulate gene expression post-transcriptionally, are implicated in various pathological conditions including diabetes mellitus (DM). DM has been increasingly recognized as an inflammatory disease and monocytes play a key role in propagating inflammation under hyperglycemic conditions. We hypothesize that high glucose dysregulates microRNAs to promote monocyte inflammatory activity, which may contribute to the pathogenesis of DM. THP-1 monocytes were cultured in normal (5 mM) and high (25 mM) glucose conditions. RT-qPCR and Western blotting were performed to assay microRNAs and proteins, respectively. Monocytes were transfected with microRNA mimics using Lipofectamine RNAiMAX reagent. THP-1 monocyte growth was assessed using Calcein-AM dye and a Boyden chamber assay was applied to measure monocyte migration. The results showed that high glucose downregulated miR-139-5p associated with increased protein expression of CXCR4, an experimentally validated target of miR-139-5p. Correspondingly, treatment with high glucose resulted in a significant increase in THP-1 cell migration towards SDF-1, a cognate ligand for CXCR4. MiR-139-5p overexpression inhibited high glucose-induced CXCR4 expression, leading to reduced cell migration towards SDF-1. High glucose did not affect THP-1 monocyte growth. In conclusion, the miR-139-5p-CXCR4 axis may play a role in high glucose-induced inflammation by regulating monocyte migration.
微小RNA是一类在转录后调节基因表达的小非编码RNA分子，与包括糖尿病（DM）在内的各种病理状况有关。糖尿病越来越被认为是一种炎症性疾病，单核细胞在高血糖条件下的炎症传播中起关键作用。我们假设高血糖会使微小RNA失调，从而促进单核细胞的炎症活性，这可能有助于糖尿病的发病机制。THP-1单核细胞在正常（5 mM）和高（25 mM）葡萄糖条件下培养。分别进行RT-qPCR和蛋白质印迹分析微小RNA和蛋白质。使用Lipofectamine RNAiMAX试剂用微小RNA模拟物转染单核细胞。使用钙黄绿素-AM染料评估THP-1单核细胞的生长，并应用博伊登小室试验测量单核细胞迁移。结果表明，高血糖下调了miR-139-5p，这与miR-139-5p的实验验证靶点CXCR4的蛋白质表达增加有关。相应地，高糖处理导致THP-1细胞向CXCR4的同源配体SDF-1迁移显著增加。miR-139-5p过表达抑制了高糖诱导的CXCR4表达，导致细胞向SDF-1的迁移减少。高血糖不影响THP-1单核细胞的生长。总之，miR-139-5p-CXCR4轴可能通过调节单核细胞迁移在高糖诱导的炎症中起作用。
