Purpose: Vascular endothelial growth factor (VEGF) and its receptors have been implicated in the pathogenesis of diabetic complications, but the exact roles of these are not certain in diabetic cardiomyopathy. We examined the effects of anti-VEGF-R2 (anti-flk-1) on diabetic mice. Methods: Male non-diabetic db/m mice and diabetic db/db mice were injected with A7R, a selective peptide inhibiting VEGF-R2, 20 mg/kg three times a week for 12 weeks. Echocardiography, immunohistochemical assessment of connective tissue growth factor (CTGF), platelet endothelial cell adhesion molecule-1 (PECAM-1) and thrombospondin-1, a natural inhibitor of neovascularization was done. In addition, cardiac VEGF, Flt-1 (VEGF-R1) and Flk-1 m-RNA levels were assessed using reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Echocardiogram showed LV wall thickness was increased but LV dimension was decreased in db/db mice compared with db/m mice. LV Fractional shortening was increased in anti-Flk-1 db/db mice compared with other group. Perivascular fibrosis was observed in anti-Flk-1 db/db mice, but was not significant. CTGF and thrombospondin-1 expression were increased, but PECAM-1 was decreased in immunohistochemical staining of anti-Flk-1 db/db mice compared with other group. Flk-1 m-RNA was decreased in anti-Flk-1 db/db mice compared with db/db control mice. Conclusions: In spite of preserving LV systolic function, microvascular homeostasis was deteriorated through myocardial fibrosis and decreased neovascularization when blocking the VEGF-R2 in diabetic cardiomyopathy.