Objective To examine the association of physical activities (PA) with diabetic retinopathy (DR) progression based on a 10-year follow-up of a large cohort of working-aged diabetic populations in Australia. Methods Nine thousand and eighteen working-aged diabetic patients were enrolled from the baseline of the 45 and Up Study from New South Wales, Australia. Self-reported PA collected by questionnaire at baseline in 2006 was graded into low ( Results In the fully adjusted model, higher PA level was significantly associated with a lower risk of RPC incident (Cox-regression, p-value for trend = 0.002; medium vs. low, hazard ratio (HR) = 0.78, 95% Confidence Interval (CI): 0.61-0.98; high vs. low, HR = 0.61, 95%CI: 0.36-0.84. In addition, gender, body mass index, insulin treatment, family history of diabetes, history of cardiovascular disease were significant effect modifiers for the association between PA and RPC. Conclusions Higher PA level was independently associated with a lower risk of DR progression among working-aged diabetic populations in this large cohort study.