To determine whether an intravitreal injection of anti-vascular endothelial growth factor (anti-VEGF) in eyes with diabetic macular edema (DME) affects the vascular infarction-related molecules (VIRMs). Nineteen eyes with DME were treated with 0.5 mg of intravitreal ranibizumab (IVR), and 22 eyes with DME were treated with 2 mg of intravitreal aflibercept (IVA). Blood was collected before, 1 week and 1 month after the injections. Aqueous humor was collected before and 1 month after the injections. The concentration of the VIRMs (cardiac myoglobin, cardiac troponin, intercellular adhesion molecule, monocyte chemotactic protein-1, matrix metalloproteinase-8, placental growth factor [PlGF], tenascin-C, tissue inhibitor of metalloproteinase-1, thrombospondin-2, vascular cell adhesion molecule-1, and VEGF) were determined by the multiplex assay. After the single injection of both types of anti-VEGF agents, the concentration of aqueous VEGF decreased significantly (P < 0.01). The plasma VEGF was reduced significantly at 1 week after the IVA (93.7 ± 17.6 to 39.5 ± 11.6 pg/ml; P < 0.01) but no significant change was seen after IVR (120.2 ± 11.3 to 137.4 ± 17.7 pg/ml). No significant changes were detected for the other VIRMs in the plasma and aqueous. A single intravitreal injection of anti-VEGF for DME does not significantly affect the concentration of several VIRMs.