Vascular endothelial growth factor accelerates healing of foot ulcers in diabetic rats via promoting M2 macrophage polarization

• Published in: Diabetic medicine 2024-09, Vol.41 (9), p.e15388-n/a
• Main Authors: Liu, Fei, Xu, Xianrui, Sun, Tao
• Format: Article
• Language: English
• Subjects: Angiogenesis; Animal models; Animals; Axitinib - pharmacology; Axitinib - therapeutic use; Cell culture; Cell Movement - drug effects; Cell viability; Cholecystokinin; Collagen; Diabetes; Diabetes mellitus; Diabetes Mellitus, Experimental - complications; Diabetes Mellitus, Experimental - metabolism; Diabetic Foot - metabolism; Diabetic Foot - pathology; diabetic foot ulcer; fibroblasts; Fibroblasts - drug effects; Fibroblasts - metabolism; Foot diseases; Humans; Immunofluorescence; Indazoles - pharmacology; Indazoles - therapeutic use; Leukocyte migration; Macrophage Activation - drug effects; Macrophages; Macrophages - metabolism; Male; Phenotypes; Plantar ulcers; Polarization; Rats; Rats, Sprague-Dawley; Streptozocin; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - metabolism; Vascular endothelial growth factor receptors; VEGF; Wound healing; Wound Healing - drug effects; Wound Healing - physiology
• ISSN: 0742-3071; 1464-5491; 1464-5491
• DOI: 10.1111/dme.15388

Aim The objective was to investigate the specific role and the regulatory mechanism of vascular endothelial growth factor (VEGF) during wound healing in diabetic foot ulcer (DFU). Methods Streptozotocin‐induced diabetic rats were used to establish a DFU animal model. VEGF and Axitinib (a specific inhibitor of VEGFR) were used for treatment in vivo. The wounds at different time points were imaged and histological analysis of the wounds were performed by haematoxylin and eosin (H&E) staining and Masson's trichrome staining. Immunohistochemical staining was conducted to examine CD31 and eNOS expression in the wounds. Immunofluorescence assay and quantitative real‐time PCR were performed to examine macrophage markers. In addition, THP‐1 was differentiated to macrophages, and then treated with interleukin (IL)‐4 to induce M2 macrophages, followed by VEGF treatment. The conditional medium (CM) from VEGF‐mediated macrophages were collected to culture human dermal fibroblasts (HDFs). Cell viability and migration were measured by Cell Counting Kit (CCK)‐8, wound‐healing and Transwell assays, respectively. Results VEGF treatment remarkably accelerated wound healing of DFU rats. VEGF promoted collagen deposition and elevated CD31 and eNOS expression, confirming the pro‐angiogenesis of VEGF around diabetic wound in rats. Meanwhile, VEGF restricted pro‐inflammatory cytokines and increased F4/80 and CD206 expression, highlighting the activated macrophages and enhanced M2 macrophages following VEGF treatment in diabetic wounds of DFU rats. However, Axitinib exerted an opposite function to VEGF in DFU rats. Moreover, VEGF directly promoted macrophage polarization toward M2 phenotype in vitro, and the CM from VEGF‐mediated M2 macrophages markedly promoted HDFs proliferation, migration and collagen deposition. Conclusion VEGF might accelerate the wound healing of DFU through promoting M2 macrophage polarization and fibroblast migration.