Enumeration and functional investigation of endothelial progenitor cells in neovascularization of diabetic foot ulcer rats with a Chinese 2-herb formula

Abstract

Backbround: We investigated the effect of a Chinese 2-herb formula (NF3) on the enumeration and angiogenic differentiation of endothelial progenitor cells (EPCs) in diabetic foot ulcer rats. Methods: EPCs and stromal cell-derived factor-1α (SDF-1α) were quantified by flow cytometry and ELISA, respectively. In vitro angiogenesis assays included proliferation, adhesion, migration and tube formation. Results: Our result demonstrated that NF3 (0.98g/kg) could significantly enhance the circulating CD34+/VEGFR2+/CD45- EPCs levels in diabetic foot ulcer rats by 60% (P<0.05) through the partial elevation of SDF-1α, restoring the mobilization ability of EPCs for wound neovascularization. We successfully isolated the BM-derived EPCs to study their angiogenic potential after NF3 treatment. BM-derived EPCs significantly expressed cell surface markers of CD34, CD146 and VEGFR2 (P<0.05-0.01). NF3 could significantly stimulate the proliferation and attachment ability of EPCs dose-dependently (P<0.01-0.001). Besides, NF3 could significantly augment EPCs migration (P<0.001) and tube formation (P<0.01-0.001). Conclusions: NF3 modulated diabetic wound healing through regulation of systemic EPCs level and increase in local vascular formation.