Anti-Inflammatory Effects of Fish Oils
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1 Anti-Inflammatory Effects of Fish Oils A. Macrophage Production of Eicosanoids B. w-3 PUFA Supplementation of Macrophages C. w-3 PUFA in IR & diabetes Oswald Quehenberger Departments of Medicine and Pharmacology School of Medicine University of California San Diego National Lipid Association, 2013 Annual Scientific Session, Las Vegas
2 Eicosanoids Contribute to the Inflammatory Response Microbial Insult/Injury Initiation/Propagation of Inflammation Resolution of Inflammation Extravasation/ Recruitment Block Recruitment Non-phlogistic Phagocytosis STOP Vasodilation Prostaglandins Leukotrienes Lipoxins Resolvins Protectins w-6 Fatty Acids 20:4 and related COX LOX LOX w-3 Fatty Acids 20:5 22:6
3 Eicosanoid Signaling Pathways in Macrophages ATP ATP LPS (KLA) Eicosanoid metabolites
4 Basic Macrophage Experimental Scheme Macrophages KLA (TLR4) Eicosanoid enzyme mrna & protein ATP (P2X 7 ) Isolate Media & Solid Phase Extraction (Eicosanoids) LC-MS/MS Quehenberger et al. J.Lipid Res. 51, (2010)
5 Eicosanoid Analytes 184 Total AA OA LA LLA DGLA EPA DHA Internal Standards
6 Cellular Eicosanoid Metabolism COX CYP( ) 5-LOX CYP(EET) 12-LOX 15-LOX Buczynski, et al., J.Lipid Res. 50, (2009)
7 Cellular Eicosanoid Metabolism COX
8 Eicosanoid Genes to Metabolites Quehenberger & Dennis, N Engl J Med, 365, (2011)
9 The Beneficial Lipids in Fish Oil Fish oil is composed primarily of two w-3 PUFAs: ω 3 ω ω 3 ω Eicosapentaenoic Acid (EPA) (C20:5, -3) Docosahexaenoic Acid (DHA) (C22:6, -3) Humans synthesize from essential fatty acid α-linolenic acid (C18:3, ω-3) EPA is found at barely detectable levels in humans DHA is more abundant than EPA; found primarily in the brain and retina
10 Arachidonic Acid Serves as a Precursor for Many Eicosanoids???? EPA/DHA??? Are eicosanoids less pro-inflammatory when derived from EPA/DHA?
11 Effects of PUFA Supplementation on Fatty Acid Membrane Distribution 4000 Membrane PUFA Composition Membrane PUFA Composition PUFA (pmol/1e6 cells) PUFA Release (2 Hr LPS) AA (20:4) AdA (22:4) EPA (20:5) DPA (22:5) DHA (22:6)
12 Effects of PUFA Supplementation on Fatty Acid Release LPS ATP
13 Effects of PUFA Supplementation on TLR-4 Stimulated Eicosanoid Signaling Fold Increase Fold Decrease Not detected
14 Effects of PUFA Supplementation on TLR-4 Stimulated COX-2 Signaling Norris PC et al., PNAS 2012
15 Effects of PUFA Supplementation on ATP Stimulated Eicosanoid Signaling Fold Increase Fold Decrease Not detected
16 Effects of PUFA Supplementation on ATP Stimulated Eicosanoid Signaling Norris PC et al., PNAS 2012
17 Effect of Fatty Acids on Adipose Inflammation and IR Adopted from Glass CK and Olefsky JM Cell Metabolism 2012 Oh DY et al., Cell 2010
18 Eicosanoids in primary adipocyte conditioned medium pmol/100mg 2000 Free AA PGE2 4 2 WT GPR120KO TNF DHA TNF DHA pmol/100mg d-PGJ TNF DHA TNF DHA LTB PGF2a TNF DHA
19 w-3 Fatty Acids Reduce Inflammatory Stress in Adipose Tissue iso PGF2a III 400 Free AA pmol/mg Adipose Tissue PGD2 NC HFD w HETE NC HFD w LTB4 0 NC HFD w-3 0 NC HFD w-3 0 NC HFD w-3
20 Aspirin-Triggered Lipoxin Formation Downregulate: Neutrophil transmigration Cytokine release Promote resolution: Removal of apoptotic cells by macs Antifibrotic Signals through ALX (FPR2) Adopted from Serhan et al. Annu.Rev.Pathol. Mech.Dis Ye et al. BBRC 1992
21 Omega-3 Fatty Acids Inhibit COX-derived 15-HETE
22 Global Summary of Fish Oil Effects Supplement Control AA EPA DHA Membrane AA AA AdA AA EPA DPA AA DHA PLA 2 AA AA AdA AA EPA DPA AA DHA TLR-4 P2X 7 TLR-4 P2X 7 TLR-4 P2X 7 TLR-4 P2X 7 COX 2 COX 1 5 LOX COX 2 COX 1 5 LOX COX 2 COX 1 5 LOX COX 2 COX 1 5 LOX PG 2 5-HETE PG 2 5-HETE PG 2 5-HETE PG 2 5-HETE PG 3 Dihomo- PG 2 LT 4 5-HEPE PG 3 Dihomo- PG 2 LT 4 PG 3 LT 4 PG 3 LT 4 5-HEPE Dihomo- 5-HEPE Dihomo- 5-HEPE PG 2 PG 2 4/7-HDoHE Decrease No Change Increase Norris PC et al., PNAS 2012
23 Conclusions on Omega-3 Study The global effects of EPA and DHA on normal lipid metabolism can be quantitatively studied. EPA and DHA affect the overall eicosadome decreasing production of some, but not all, AAderived eicosanoids. There is a concomitant increase in specific EPAand DHA-derived metabolites. cpla2 releases EPA/DHA from membrane phospholipids. Deciphering the role of fish oil-derived ω-3 EPA and DHA in inflammatory eicosanoid signaling provides insight as to their role as therapeutic agents in human disease.
24 Acknowledgements UCSD Paul C. Norris Darren Dumlao Aaron M. Armando Ishita Shah Edward A. Dennis, UCSD Jerrold M. Olefsky, UCSD Da Young Oh, UCSD Ruedi Aebersold, ETH Zurich Eduard Sabido, Center of Genomics Regulation Barcelona LIPID MAPS Large Scale Collaborative Grant GM from the National Institutes of Health
25
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