Diabetic retinopathy may induce visual impairment. We evaluated vision-related quality of life in patients with visual acuity <5/10 in the better eye induced by retinopathy using the 25-item National Eye Institute Visual Functioning Questionnaire (NEI VFQ-25). The NEI VFQ-25 was self-administered to 196 patients in 3 Italian centres (A, B and C; n = 64, 61 and 71, respectively) dedicated to DR screening and treatment. Patients in the 3 centres did not differ by age, gender, occupation and diabetes duration. Multivariate analysis demonstrated that reduced visual acuity was associated with decreased scores for General Vision, Near Activities, Distance Activities, Visual-Specific Social Functioning, Mental Health, Role Difficulties and Dependency, Driving, Colour Vision and Peripheral Vision (p < 0.01, all). Treatment by photocoagulation was associated with reduced scores in General Health (-8.3; p = 0.002), General Vision (-7.2; p = 0.001), Visual-Specific Role Difficulties (-8.8; p = 0.015) and Driving (-13.7; p = 0.003). Centre affiliation was associated with different scores for General Health, Ocular pain, Distance Activities, Visual-Specific Social Functioning and Role Difficulties and Peripheral Vision. Women had higher scores for General Vision (p = 0.015), Near Activities (p = 0.005), Distance Activities (p = 0.006), Visual-Specific Social Functioning (p = 0.03), Visual-Specific Mental Health (p = 0.035) and Colour Vision (p = 0.012). Diabetic retinopathy and vision loss modify the way people perceive their own ability to function autonomously. More data should be collected to confirm this interpretation and to guide the development of more appropriate settings to improve approach and support to patients.