title: Combination of n-3 polyunsaturated fatty acids reduces atherogenesis in apolipoprotein E-deficient mice by inhibiting macrophage activation.
abstract: Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are major components of n-3 polyunsaturated fatty acids (n-3 PUFAs) which inhibit atherogenesis, although few studies have examined the effects of the combination of EPA and DHA on atherogenesis. The aim of this study was to investigate whether DHA has additional anti-atherosclerotic effects when combined with EPA.
  
  
  
  Male 8-week-old apolipoprotein E-deficient (Apoe-/-) mice were fed a western-type diet supplemented with different amounts of EPA and DHA; EPA (2.5%, w/w), low-dose EPA + DHA (2.5%, w/w), or high-dose EPA + DHA (5%, w/w) for 20 weeks. The control group was fed a western-type diet containing no n-3 PUFA. Histological and gene expression analysis were performed in atherosclerotic lesions in the aorta. To address the mechanisms, RAW264.7 cells were used.
  
  
  
  All n-3 PUFA treatments significantly attenuated the development and destabilization of atherosclerotic plaques compared with the control. The anti-atherosclerotic effects were enhanced in the high-dose EPA + DHA group (p < 0.001), whereas the pure EPA group and low-dose EPA + DHA group showed similar results. EPA and DHA additively attenuated the expression of inflammatory molecules in RAW264.7 cells stimulated with LPS. DHA or EPA + DHA suppressed LPS-induced toll-like receptor 4 (TLR4) expression in lipid rafts on RAW264.7 cells (p < 0.05). Lipid raft disruption by methyl-β-cyclodextrin suppressed mRNA expression of inflammatory molecules in LPS-stimulated macrophages.
  
  
  
  n-3 PUFAs suppressed atherogenesis. DHA combined with EPA had additional anti-inflammatory effects and inhibited atherogenesis in Apoe-/-mice. The reduction of TLR4 expression in lipid rafts in macrophages by DHA might be involved in this mechanism, at least partially.
authors.affiliation: Department of Cardiovascular Medicine, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Takashima
authors.affiliation: Department of Cardiovascular Medicine, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan. Electronic address: daiju.fukuda@tokushima-u.ac.jp.
authors.forename: Daiju
authors.initials: D
authors.lastname: Fukuda
authors.affiliation: Division for Health Service Promotion, The University of Tokyo, Tokyo 113-0033, Japan.
authors.forename: Kimie
authors.initials: K
authors.lastname: Tanaka
authors.affiliation: Department of Cardiovascular Medicine, The University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Yasutomi
authors.initials: Y
authors.lastname: Higashikuni
authors.affiliation: Department of Pediatrics, The University of Tokyo Hospital, Tokyo 113-8655, Japan.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Hirata
authors.affiliation: Department of Nutrition and Metabolism, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Nishimoto
authors.affiliation: Department of Cardiovascular Medicine, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
authors.forename: Shusuke
authors.initials: S
authors.lastname: Yagi
authors.affiliation: Department of Cardiovascular Medicine, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
authors.forename: Hirotsugu
authors.initials: H
authors.lastname: Yamada
authors.affiliation: Department of Cardiovascular Medicine, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Soeki
authors.affiliation: Department of Cardiovascular Medicine, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
authors.forename: Tetsuzo
authors.initials: T
authors.lastname: Wakatsuki
authors.affiliation: Department of Clinical Nutrition and Food Management, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Taketani
authors.affiliation: Department of Cardio-Diabetes Medicine, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
authors.forename: Michio
authors.initials: M
authors.lastname: Shimabukuro
authors.affiliation: Department of Cardiovascular Medicine, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
authors.forename: Masataka
authors.initials: M
authors.lastname: Sata
mesh_terms: D000818:Animals; D001012:Aorta, Abdominal / Q000473:pathology; D050197:Atherosclerosis / Q000378:metabolism / Q000517:prevention & control / Q000628:therapy*; D001794:Blood Pressure; D004281:Docosahexaenoic Acids / Q000494:pharmacology; D015118:Eicosapentaenoic Acid / Q000494:pharmacology; D015525:Fatty Acids, Omega-3 / Q000378:metabolism*; D007249:Inflammation; D008055:Lipids / Q000737:chemistry; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000378:metabolism; D008297:Male; D021962:Membrane Microdomains / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D000074085:Mice, Knockout, ApoE; D000067996:RAW 264.7 Cells
chemical_list: D015525:Fatty Acids, Omega-3; D008055:Lipids; D004281:Docosahexaenoic Acids; D015118:Eicosapentaenoic Acid
keywords: Atherosclerosis; Inflammation; Lipid raft; Macrophage; Toll-like receptor 4; n-3 poly unsaturated fatty acid