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Epidemiologic and clinical studies have suggested that hormone therapy (HT) and n-3 polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), exert antidepressant effects through regulation of the serotonergic pathway. The present study was conducted to evaluate the hypothesis that erythrocyte levels of n-3 PUFAs and depression were negatively associated in postmenopausal women, particularly those using HT.A total of 214 postmenopausal women were divided into groups defined by depression and HT status. Depression was defined as a Beck Depression Inventory score of at least 14, and the types and durations of HT were obtained from medical records.Erythrocyte levels of EPA, docosapentaenoic acid (DPA), DHA, omega-3 index (sum of EPA + DHA), total n-3 PUFA, and arachidonic acid were significantly higher in women using HT than in those not using HT. There were significant negative correlations between depression and erythrocyte levels of alpha-linolenic acid (r = −0.310), DPA (r = −0.272), DHA (r = −0.209), and total n-3 PUFA (r = −0.234) in women using HT. Multivariate-adjusted logistic regression analysis also showed that depression was negatively associated with the erythrocyte levels of alpha-linolenic acid (P for trend = 0.018), DPA (P for trend = 0.014), and DHA (P for trend = 0.036), only in women using HT.Negative associations between erythrocyte levels of n-3 PUFAs and depression were found in Korean postmenopausal women using HT but not in those not using HT, suggesting a synergistic effect of HT and n-3 PUFAs on depression.