Research Paper Volume 13, Issue 1 pp 1186—1211

MicroRNA-139-5p upregulation is associated with diabetic endothelial cell dysfunction by targeting c-jun

Yu-Fang Luo1, *, , Xin-Xing Wan1, *, , Li-Ling Zhao1, *, , Zi Guo1, , Rui-Ting Shen2, , Ping-Yu Zeng3, , Ling-Hao Wang1, , Jing-Jing Yuan1, , Wen-Jun Yang1, , Chun Yue1, , Zhao-Hui Mo1, ,

  • 1 Department of Endocrinology, Third Xiangya Hospital of Central South University and Diabetic Foot Research Center of Central South University, Changsha 410013, Hunan Province, China
  • 2 Key Laboratory of Hormones and Development (Ministry of Health), Tianjin Key Laboratory of Metabolic Diseases, Tianjin Metabolic Diseases Hospital and Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300070, China
  • 3 Center of Experimental Medicine, Third Xiangya Hospital of Central South University, Changsha 410013, Hunan Province, China
* Equal contribution

Received: May 30, 2020       Accepted: October 3, 2020       Published: December 3, 2020      

https://doi.org/10.18632/aging.202257
How to Cite

Copyright: © 2020 Luo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Dysfunction of endothelial cells (ECs) and their progenitor cells is an important feature of diabetic vascular disease. MicroRNA (miR)-139-5p is involved in inhibiting the metastasis and progression of diverse malignancies. However, the role of miR-139-5p in ECs still remains unclarified. Here we demonstrated that miR-139-5p expression was elevated in endothelial colony-forming cells (ECFCs) isolated from patients with diabetes, ECs derived from the aorta of diabetic rodents, and human umbilical vein endothelial cells (HUVECs) cultured in high glucose media. MiR-139-5p mimics inhibited tube formation, migration, proliferation, and down-regulated expression of c-jun, vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF)-B, in ECFCs and HUVECs, respectively; moreover, miR-139-5p inhibitors reversed the tendency. Further, gain- and-loss function experiments and ChIP assay indicated that miR-139-5p regulate functions of ECFCs by targeting c-jun-VEGF/PDGF-B pathway. In vivo experiments (Matrigel plug assay and hindlimb ischemia model) showed that miR-139-5p downregulation further promoted ECFC-mediated angiogenesis and blood perfusion. In conclusion, diabetes-mediated high miR-139-5p expression inhibits the c-jun-VEGF/PDGF-B pathway, thus decreasing ECFCs migration, tube formation and proliferation, which subsequently reduces ECs survival. Therefore, miR-139-5p might be an important therapeutic target in the treatment of diabetic vasculopathy in the future.

Abbreviations

AP-1: activator protein-1; CD31: cluster of differentiation 31; ChIP: chromatin immunoprecipitation; CLI: critical limb ischemia; DAB: 3:3′-diaminobenzidine; DMEM: Dulbecco’s modified Eagle’s medium; EC: endothelial cell; ECFC: endothelial colony-forming cell; EDTA: ethylenediaminetetraacetic acid; EGM-2: endothelial cell growth medium 2; EPC: endothelial progenitor cell; FBS-DMEM: fetal bovine serum-Dulbecco’s modified Eagle’s medium; HLI: hind limb ischemia; HPP: high proliferation potential; HUVEC: human umbilical vein endothelial cell; LPP: low proliferation potential; miR: micro-ribonucleic acid; MSC: mesenchymal stem cell; NF-κB: nuclear factor-kappa-light-chain-enhancer of activated B cells; PBS: phosphate-buffered saline; PCR: polymerase chain reaction; PDGF: platelet-derived growth factor; PGC-1α: peroxisome proliferator-activated gamma coactivator 1-alpha; PKC: protein kinase C; SDS: sodium dodecyl sulfate; siRNA: small interfering RNA; STZ: streptozotocin; VEGF: vascular endothelial growth factor; vWF: von Willebrand factor; α-SMA: anti-alpha smooth muscle actin.