Oxidative stress-induced endothelial cells-derived exosomes accelerate skin flap survival through Lnc NEAT1-mediated promotion of endothelial progenitor cell function

• Published in: Stem cell research & therapy Vol. 13; no. 1; pp. 1 - 325
• Main Authors: Guo, Linlin, Chen, Yuxuan, Feng, Xiaoling, Sun, Di, Sun, Jiaming, Mou, Shan, Zhao, Kangcheng, An, Ran
• Format: Journal Article
• Language: English
• Published: London BioMed Central Ltd 18.07.2022; BioMed Central; BMC
• Subjects: Analysis; Angiogenesis; Antibodies; Apoptosis; Cell culture; DNA microarrays; Endothelial cells; Endothelial progenitor cells; Endothelium; Exosomes; Flow cytometry; Gene expression; Gene silencing; Genetic engineering; Hydrogen peroxide; Instrument industry; Lnc NEAT1; Non-coding RNA; Oxidative stress; Photographic industry; Polymerase chain reaction; Progenitor cells; Proteins; Reconstructive surgery; Signal transduction; Skin; Skin flap; Stem cells; Surgery, Plastic; Transplantation; Umbilical cord; Western blotting; Wnt protein; β-Catenin; United Kingdom; Australia; United States; Japan; China; Germany; Beijing China; United States > US
• ISSN: 1757-6512; 1757-6512
• DOI: 10.1186/s13287-022-03013-9

Background Flap transplantation is commonly used in reconstructive surgery. A prerequisite for skin flap survival is sufficient blood supply. However, such approaches remain unclear. This study aimed to explore the underlying mechanisms of exosomes derived from human umbilical vascular endothelial cells (HUVECs) exposed to oxidative stress on endothelial progenitor cells (EPCs) and their subsequent influence on the survival of skin flaps. Methods HUVECs were treated with various concentrations of H.sub.2O.sub.2 to establish an oxidative stress model. To investigate the effects of H.sub.2O.sub.2-HUVEC-Exos and HUVEC-Exos, Cell Counting Kit-8, tube formation, invasion assays, and quantitative real-time polymerase chain reaction (qRT-PCR) were performed in EPCs. Microarray analysis was used to reveal the differentially expressed long non-coding RNAs (lncRNAs) in the H.sub.2O.sub.2-HUVEC-Exos and HUVEC-Exos. In addition, gene silencing and western blotting were employed to determine the mechanism behind lncRNA nuclear enrichment enriched transcript 1 (Lnc NEAT1) in EPCs. Further, a rat skin flap model was used to determine the role of the exosomes in skin flap survival in vivo. Results HUVECs were stimulated with 100 [mu]mol/L H.sub.2O.sub.2 for 12 h to establish an oxidative stress model. H.sub.2O.sub.2-HUVEC-Exos promoted the proliferation, tube formation, and invasion of EPCs and remarkably increased skin flap survival compared to the HUVEC-Exos and control groups. Sequencing of exosome RNAs revealed that the Lnc NEAT1 level was dramatically increased in the H.sub.2O.sub.2-HUVEC-Exos, leading to activation of the Wnt/[beta]-catenin signaling pathway. Comparatively, knockdown of Lnc NEAT1 in HUVEC-Exos and H.sub.2O.sub.2-HUVEC-Exos significantly inhibits the angiogenic capacity of EPCs, reduced the survival area of skin flap and downregulated the expression levels of Wnt/[beta]-catenin signaling pathway proteins, whereas Wnt agonist partly reversed the negative effect of NEAT1 downregulation on EPCs through the Wnt/[beta]-catenin signaling pathway. Conclusions Exosomes derived from HUVECs stimulated by oxidative stress significantly promoted the pro-angiogenic ability of EPCs through the Wnt/[beta]-catenin signaling pathway mediated by Lnc NEAT1 and hence enhanced random flap survival in vivo. Therefore, the application of H.sub.2O.sub.2-HUVEC-Exos may serve as an alternative therapy for improving random skin flap survival. Keywords: Oxidative stress, Exosomes, Endothelial progenitor cells, Angiogenesis, Lnc NEAT1, Skin flap