The aim of this study was to evaluate the effect of kale (Brassica oleracea acephala) juice supplementation on serum lipid levels and phospholipid (PL) fatty acid compositions in hypercholesterolemic men. Thirty-two men with hypercholesterolemia (>200 mg/dl) were recruited among the faculty and staff at Y University after annual health examinations. The subjects consumed 150 ml of kale juice per day for a 12-week intervention period. Dietary and anthropometric assessments were performed before and after supplementation, respectively, to ensure that the subjects maintained their usual diet and lifestyle throughout the intervention. Serum concentrations of HDL-cholesterol and the ratio of HDL-cholesterol to LDL-cholesterol were significantly increased (p<0.001) after intervention. Serum LDL-cholesterol concentration and atherogenic index were significantly reduced (p<0.001). Levels of 12:0, 14:0, $18:1{omega}9$, $18:3{omega}6$, and sum of monounsaturated fatty acids (MUFA) in serum pholspholipid (PL) were significantly increased, while $22:4{omega}6$ level was significantly reduced (p<0.05). It can be speculated that kale juice, containing large amounts of antioxidant nutrients, contributes to changes of serum-PL-fatty acid compositions and the improvements of serum lipid profiles. This study demonstrates the supplementation of regular meals with kale juice may favorably affect serum lipid profiles and serum-PL fatty acid compositions and, hence, could lower the risks of coronary artery disease in men with hypercholesterolemia.