An elevated level of non-high-density lipoprotein cholesterol (non-HDL-C) is a major risk factor for cardiovascular disease. The purpose of this study was to use the meta-analytic approach to examine the effects of walking on non-HDL-C in adults. Twenty-two randomized controlled trials representing 30 outcomes from 948 subjects (573 exercise, 375 control) met our inclusion criteria. Across all designs and categories, random effects modeling resulted in a significantly greater decrease in the walking group when compared with the control group of approximately 4% for non-HDL-C (+/- standard error of the mean, -5.6+/-1.8 mg/dL, 95% confidence interval, -8.8 to -2.4 mg/dL). Meta-regression showed a statistically significant association between changes in non-HDL-C and the year of publication, with greater reductions associated with more recent publication year (R2 = 0.23, p = 0.005). The results of this meta-analytic review suggest that walking reduces non-HDL-C in adult humans.