High Glucose-Induced Senescent Fibroblasts-Derived Exosomal MiR-497 Inhibits Wound Healing by Regulating Endothelial Cellular Autophagy Via ATG13

Overview

Journal Anal Cell Pathol (Amst)

Date 2025 Jan 20

PMID 39831298

Authors

Changjiang Liu

Yuting Liu

Yifeng Yu

Siyuan Huang

Chao Sun

Dong Zhang

Aixi Yu

Affiliations

Soon will be listed here.

Abstract

Fibroblasts play a crucial role in diabetic wound healing, and their senescence is the cause of delayed wound repair. It was reported that fibroblasts can secrete exosomes that can mediate a vital role in diabetic complications. Our purpose is to examine the biological function of high glucose (HG)-induced senescent fibroblasts from the perspective of exosomes and reveal the mechanism at cellular and animal levels. HG-induced senescent fibroblasts were measured by senescence-associated -galactosidase staining and immunofluorescence. Flow cytometry, 5-ethynyl-2'-deoxyuridine (edu), and cell counting kit 8 (CCK-8) assay were applied to detect apoptosis and cell viability. Fibroblasts and endothelial cells were cocultured, and the migration and angiogenesis abilities were detected by scratch, transwell, and tube formation assays. Exosomes were isolated and identified from fibroblasts that were treated differently. Then, the function of exosomes was investigated in cells and mice, including examining the cellular phenotype changes, detecting the autophagy levels, and evaluating the wound healing rate. Furthermore, the potential mechanism by which senescent fibroblast-derived exosomes inhibit wound healing was examined via bioinformatics, real-time quantitive polymerase chain reaction (qPCR), transfection, and dual-luciferase assays. It illustrated that HG-induced senescent fibroblasts exhibited adverse impacts on cellular proliferation, migration, and angiogenesis of endothelial cells via secreting exosomes, and senescent fibroblast-derived exosomes (S-Exos) can delay skin wound defects in mice. Subsequent differential analysis of the GSE153214 and GSE48417 datasets elucidated that miR-497 was the biomarker in the senescent fibroblasts. Interestingly, the miR-497 levels were also elevated in S-Exos. Its overexpression can regulate human umbilical vein endothelial cell function by regulating autophagy via targeting ATG13. Furthermore, experiments also illustrated that miR-497 can delay wound healing and reduce autophagy. Our study demonstrated that exosomes from senescent fibroblasts can impair endothelial cell function and impede diabetic wound healing. The underlying mechanism was that fibroblast-derived exosomal miR-497 can target ATG13 to reduce autophagy, offering insight into new therapy for diabetic complications and other diseases.

References

Lima A, Illing T, Schliemann S, Elsner P . Cutaneous Manifestations of Diabetes Mellitus: A Review. Am J Clin Dermatol. 2017; 18(4):541-553. DOI: 10.1007/s40257-017-0275-z. View

Spravchikov N, Sizyakov G, Gartsbein M, Accili D, Tennenbaum T, WERTHEIMER E . Glucose effects on skin keratinocytes: implications for diabetes skin complications. Diabetes. 2001; 50(7):1627-35. DOI: 10.2337/diabetes.50.7.1627. View

Dow C, Mancini F, Rajaobelina K, Boutron-Ruault M, Balkau B, Bonnet F . Diet and risk of diabetic retinopathy: a systematic review. Eur J Epidemiol. 2017; 33(2):141-156. DOI: 10.1007/s10654-017-0338-8. View

Chen X, Lin S, Gu L, Zhu X, Zhang Y, Zhang H . Inhibition of miR-497 improves functional outcome after ischemic stroke by enhancing neuronal autophagy in young and aged rats. Neurochem Int. 2019; 127:64-72. DOI: 10.1016/j.neuint.2019.01.005. View

Wu T, Qiao S, Shi C, Wang S, Ji G . Metabolomics window into diabetic complications. J Diabetes Investig. 2017; 9(2):244-255. PMC: 5835462. DOI: 10.1111/jdi.12723. View

Gualdi G, Crotti S, Monari P, Calzavara-Pinton P, Vitali M, Baronio M . The nested graft acts by inducing the process of de-senescence of the fibroblasts in chronic venous ulcers. Int Wound J. 2015; 13(6):1104-1110. PMC: 7949948. DOI: 10.1111/iwj.12415. View

Wu Z, Cai X, Huang C, Xu J, Liu A . miR-497 suppresses angiogenesis in breast carcinoma by targeting HIF-1α. Oncol Rep. 2016; 35(3):1696-702. DOI: 10.3892/or.2015.4529. View

Zhou X, Guo Y, Yang K, Liu P, Wang J . The signaling pathways of traditional Chinese medicine in promoting diabetic wound healing. J Ethnopharmacol. 2021; 282:114662. DOI: 10.1016/j.jep.2021.114662. View

Soydas T, Sarac E, Cinar S, Dogan S, Solakoglu S, Tuncdemir M . The protective effects of metformin in an in vitro model of aging 3T3 fibroblast under the high glucose conditions. J Physiol Biochem. 2018; 74(2):273-281. DOI: 10.1007/s13105-018-0613-5. View

10.

Wang Y, Gao Y, Huang Y, Pan Y, Yu Y, Zhou Y . The potential protective effects of miR-497 on corneal neovascularization are mediated via macrophage through the IL-6/STAT3/VEGF signaling pathway. Int Immunopharmacol. 2021; 96:107745. DOI: 10.1016/j.intimp.2021.107745. View

11.

Berlanga-Acosta J, Mendoza-Mari Y, Martinez M, Valdes-Perez C, Ojalvo A, Armstrong D . Expression of cell proliferation cycle negative regulators in fibroblasts of an ischemic diabetic foot ulcer. A clinical case report. Int Wound J. 2012; 10(2):232-6. PMC: 7950332. DOI: 10.1111/j.1742-481X.2012.12000.x. View

12.

Tkach M, Thery C . Communication by Extracellular Vesicles: Where We Are and Where We Need to Go. Cell. 2016; 164(6):1226-1232. DOI: 10.1016/j.cell.2016.01.043. View

13.

Zhu W, Zhang H, Gao J, Xu Y . Silencing of miR-497-5p inhibits cell apoptosis and promotes autophagy in Parkinson's disease by upregulation of FGF2. Environ Toxicol. 2021; 36(11):2302-2312. DOI: 10.1002/tox.23344. View

14.

Ren H, Zhao F, Zhang Q, Huang X, Wang Z . Autophagy and skin wound healing. Burns Trauma. 2022; 10:tkac003. PMC: 8847901. DOI: 10.1093/burnst/tkac003. View

15.

van Deursen J . The role of senescent cells in ageing. Nature. 2014; 509(7501):439-46. PMC: 4214092. DOI: 10.1038/nature13193. View

16.

Feng L, Cheng K, Zang R, Wang Q, Wang J . miR-497-5p inhibits gastric cancer cell proliferation and growth through targeting PDK3. Biosci Rep. 2019; 39(9). PMC: 6732365. DOI: 10.1042/BSR20190654. View

17.

Kalluri R, LeBleu V . The biology function and biomedical applications of exosomes. Science. 2020; 367(6478). PMC: 7717626. DOI: 10.1126/science.aau6977. View

18.

Lee P, Halter J . The Pathophysiology of Hyperglycemia in Older Adults: Clinical Considerations. Diabetes Care. 2017; 40(4):444-452. DOI: 10.2337/dc16-1732. View

19.

Han Z, Zhang Y, Yang Q, Liu B, Wu J, Zhang Y . miR-497 and miR-34a retard lung cancer growth by co-inhibiting cyclin E1 (CCNE1). Oncotarget. 2015; 6(15):13149-63. PMC: 4537005. DOI: 10.18632/oncotarget.3693. View

20.

Wu Q, Zhou L, Lv D, Zhu X, Tang H . Exosome-mediated communication in the tumor microenvironment contributes to hepatocellular carcinoma development and progression. J Hematol Oncol. 2019; 12(1):53. PMC: 6542024. DOI: 10.1186/s13045-019-0739-0. View