Research Paper Volume 13, Issue 15 pp 19805—19821
MiR-195-5p and miR-205-5p in extracellular vesicles isolated from diabetic foot ulcer wound fluid decrease angiogenesis by inhibiting VEGFA expression
- 1 Department of Endocrinology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, China
Received: December 17, 2020 Accepted: May 31, 2021 Published: August 9, 2021https://doi.org/10.18632/aging.203393
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Copyright: © 2021 Liu 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.
Diabetic foot ulcers are recalcitrant to healing, and poor angiogenesis is considered as the main contributing factor. We aimed to explore the effect of extracellular vesicles (EVs) derived from wound fluids on new vessel formation in diabetic foot ulcers. EVs were isolated from wound fluids of diabetic foot ulcers (DF-EVs). The inhibitory effect of DF-EVs on human umbilical vein endothelial cells (HUVECs) and wound healing was tested. To elucidate the potential mechanism of these effects, we screened the differentially expressed microRNAs (miRNAs) in DF-EVs via microarray analysis and verified the upregulation of miR-195-5p and miR-205-5p in DF-EVs via quantitative real-time polymerase chain reaction (qRT-PCR). Further dual-luciferase reporter assays and overexpression experiments proved these two miRNAs inhibited the expression of vascular endothelial growth factor A (VEGFA) directly to the 3′ untranslated region (UTR) of VEGFA and, in turn, promoted an inhibitory effect of DF-EVs on angiogenesis and wound healing in patients with diabetic foot ulcers. Our study shows EVs in the wound fluids of diabetic foot ulcer lesions carrying antiangiogenic miR-195-5p and miR-205-5p negatively regulated angiogenesis and wound healing in patients with diabetic foot.
AGEs: advanced glycation end products; Control-EVs: extracellular vesicles deprived from wound fluids of nondiabetic wounds; Cr: serum creatinine; DF: diabetic foot ulcers; DF-EVs: extracellular vesicles deprived from wound fluids of diabetic foot ulcers; DM: diabetes mellitus; ECs: endothelial cells; EVs: extracellular vesicles; FBG: fasting blood glucose; HbA1C: hemoglobin A1C; HDL-C: high-density lipoprotein cholesterol; HE: hematoxylin and eosin; HEK-293T: Human embryonic kidney 293T; HRP: horseradish peroxidase; HUVECs: human umbilical vein endothelial cells; LDL-C: low-density lipoprotein cholesterol; miRNAs: microRNAs; MSCs: mesenchymal stem cells; MUT: mutated; N.C.: negative control; NTA: nanoparticle tracking analysis; qRT-PCR: quantitative real-time polymerase chain reaction; SD: standard deviation; TC: total cholesterol; TG: triglyceride; UTR: untranslated region; WT: wild type.