Fig. 4

CXCL5 neutralizing antibody repaired neovascularization and wound healing in type 1 DM mice. Serum CXCL5 levels in the diabetic mice were higher than those in the non-DM control. Treatment with CXCL5 mAb reduced CXCL5 levels (n = 6; A). Representative evaluation of the ischemic (right) and nonischemic (left) hindlimbs before, immediately after 2 weeks and 4 weeks after the hindlimb ischemia surgery in STZ induced type 1 diabetic mice (n = 6; B). The number of circulating EPCs was determined by flow cytometry in STZ induced type 1 diabetic mice. Treatment with CXCL5 mAb 100 μg increased the number of circulating EPCs after ischemia surgery compared with DM (n = 6; C). Anti-CD31 immunostaining showed that CXCL5 mAb 100 μg treatment significantly increased the number of capillaries. Scale bar, 50 µm (n = 6; D) Western blotting and statistical analyses of VEGF and SDF-1 in the ischemia leg (n = 3; E). Angiogenesis in aortic ring cultures from CXCL5 mAb 100 μg mice was significantly increased the number of vessels sprouting compared with DM mice. Scale bar, 50 µm (n = 3; F). Representative matrigel plug images and analysis of hemoglobin content (n = 6; G). Representative matrigel plug images with immunostaining of CD31. CD31 positive areas were enhanced in the CXCL5 mAb 100 μg treatment mice. Scale bar, 50 µm (H). CXCL5 mAb 100 μg treatment improved wound repair ability in STZ induced type 1 diabetic mice. Representative wound areas and the closure rates of 3-mm punch biopsies were measured (n = 6; I). Representative wound area images with immunostaining of CD31. CD31 positive areas were enhanced in the CXCL5 100 μg mAb treatment mice. Scale bar, 50 µm (J). Representative skin images with masson trichrome staining. Collagen depositions were enhanced in the CXCL5 mAb 100 μg treatment mice. Scale bar, 50 and 500 µm (K). CXCL5 Chemokine C-X-C motif ligand 5, DM diabetes mellitus, EPC endothelial progenitor cell, mAb monoclonal antibody, STZ Streptozotocin, SDF-1 stromal cell-derived factor 1, VEGF vascular endothelial growth factor. The Mann–Whitney U test was used to determine statistically significant differences. *p < 0.05, **p < 0.01 compared with the non-DM control. ^#p < 0.05, ^##p < 0.01 compared with the untreated DM group