A combination of healthy lifestyle factors is associated with lower risks of coronary heart disease, diabetes and stroke, but little is known about its association with chronic kidney disease (CKD). This study analyzed the effect of a combination of healthy lifestyle factors on the incidence of proteinuria among participants without CKD. Of the 7565 persons aged 40-79 years who participated in the Specific Health Checkups and Guidance System in Sado Island, Japan in 2008, 4902 participants (2015 males) without CKD were included. The healthy lifestyle score was calculated by summing the total number of lifestyle factors for which the participants were at low risk. Low risk was defined as (1) nonsmoker, (2) body mass index (BMI) <25 kg m(-2), (3) moderate or less alcohol consumption, (4) regular exercise and (5) better eating patterns. Logistic analysis was used to examine the relationship between the baseline score in 2008 and the development of proteinuria in 2009. Proteinuria developed in 2.2% of participants (males, 3.2; females, 1.5%). Compared with participants with a healthy lifestyle score of 0 to 2, participants with a score of 5 had a lower risk (odds ratio: 0.39, 95% confidence interval: 0.16-0.94), independently of having diabetes, hypertension and hypercholesterolemia. Overall, 47% of the cases in this cohort could be attributed to lack of adherence to this low-risk pattern. These findings underscore the importance of a healthier lifestyle in preventing CKD.