Objective To investigate the role of vascular endothelial growth factor-activating transcriptional factor (VEGF-ATF) on the VEGF signaling pathway in diabetes mellitus. Methods Totally, 20 C57BL/6 mice fed with high fat diet was induced into diabetes mellitus. Ten diabetes mellitus mice received a lower limb muscle injection with VEGF-ATF plasmid, and another ten were as control. VEGF-ATF is an engineered transcription factor designed to increase VEGF expression. Three days later, mice were sacrificed and the injected gastrocnemius was used for analysis. VEGF mRNA and protein expressions were examined by real-time PCR and ELISA respectively. VEGF receptor 2 mRNA expression was tested with RT-PCR. Phosphorylated Akt, Akt, endothelial nitric oxide synthase (eNOS), and phosphorylated eNOS were assessed by western blot. Results At 3 days post-injection, in mice with diabetes mellitus, VEGF gene transfer increased VEGF mRNA copies and VEGF protein expression in injected muscles compared with control; and reinstated the impaired VEGF signaling pathway with increasing the ratios of phosphorylated Akt/Akt and phosphorylated eNOS/eNOS. However, it did not affect the expression of VEGF receptor 2 mRNA. Conclusion Gene transfer with VEGF-ATF is able to reinstate the impaired VEGF downstream pathway, and potentially promote therapeutic angiogenesis in mice with diabetes mellitus. Objective To investigate the role of vascular endothelial growth factor-activating transcriptional factor (VEGF-ATF) on the VEGF signaling pathway in diabetes mellitus. Methods Totally, 20 C57BL / 6 mice fed with high fat diet was induced into diabetes mellitus. Ten diabetes mellitus VEGF-ATF is an engineered transcription factor designed to increase VEGF expression. Three days later, mice were sacrificed and the injected gastrocnemius was used for analysis. VEGF mRNA and protein expressions were detected by real-time PCR and ELISA respectively. VEGF receptor 2 mRNA expression was tested with RT-PCR. Phosphorylated Akt, Akt, endothelial nitric oxide synthase (eNOS), and phosphorylated eNOS were assessed by western blot. Results At 3 days post-injection, in mice with diabetes mellitus, VEGF gene transfer increased VEGF mRNA copies and VEGF protein expression in injected muscles compared wit h control; and reinstated the impaired VEGF signaling pathway with increasing the ratios of phosphorylated Akt / Akt and phosphorylated eNOS / eNOS. However, it did not affect the expression of VEGF receptor 2 mRNA. Conclusion Gene transfer with VEGF-ATF is able to reinstate the impaired VEGF downstream pathway, and potentially promote therapeutic angiogenesis in mice with diabetes mellitus.