Abstract
BackgroundIncreased consumption of omega-3 (ω-3) fatty acids found in cold-water fish and fish oil has been reported to protect against obesity. A potential mechanism may be through reduction in adipocyte differentiation. Stearidonic acid (SDA), a plant-based ω-3 fatty acid, has been targeted as a potential surrogate for fish-based fatty acids; however, its role in adipocyte differentiation is unknown. This study was designed to evaluate the effects of SDA on adipocyte differentiation in 3T3-L1 cells.Methods3T3-L1 preadipocytes were differentiated in the presence of SDA or vehicle-control. Cell viability assay was conducted to determine potential toxicity of SDA. Lipid accumulation was measured by Oil Red O staining and triglyceride (TG) quantification in differentiated 3T3-L1 adipocytes. Adipocyte differentiation was evaluated by adipogenic transcription factors and lipid accumulation gene expression by quantitative real-time polymerase chain reaction (qRT-PCR). Fatty acid analysis was conducted by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS).Results3T3-L1 cells treated with SDA were viable at concentrations used for all studies. SDA treatment reduced lipid accumulation in 3T3-L1 adipocytes. This anti-adipogenic effect by SDA was a result of down-regulation of mRNA levels of the adipogenic transcription factors CCAAT/enhancer-binding proteins alpha and beta (C/EBPα, C/EBPβ), peroxisome proliferator-activated receptor gamma (PPARγ), and sterol-regulatory element binding protein-1c (SREBP-1c). SDA treatment resulted in decreased expression of the lipid accumulation genes adipocyte fatty-acid binding protein (AP2), fatty acid synthase (FAS), stearoyl-CoA desaturase (SCD-1), lipoprotein lipase (LPL), glucose transporter 4 (GLUT4) and phosphoenolpyruvate carboxykinase (PEPCK). The transcriptional activity of PPARγ was found to be decreased with SDA treatment. SDA treatment led to significant EPA enrichment in 3T3-L1 adipocytes compared to vehicle-control.ConclusionThese results demonstrated that SDA can suppress adipocyte differentiation and lipid accumulation in 3T3-L1 cells through down-regulation of adipogenic transcription factors and genes associated with lipid accumulation. This study suggests the use of SDA as a dietary treatment for obesity.
Highlights
Increased consumption of omega-3 (ω-3) fatty acids found in cold-water fish and fish oil has been reported to protect against obesity
Increased attention has been focused on dietary ω-3 polyunsaturated fatty acids (PUFAs), mainly EPA (20:5; ω-3) and DHA (22:6; ω-3), which have been reported to protect against the development of obesity and reduce body fat in humans [2, 3]
Effect of Stearidonic acid (SDA) on 3T3-L1 cell viability The concentration of 200 μM of ω-3 fatty acids were used in our experiments
Summary
Increased consumption of omega-3 (ω-3) fatty acids found in cold-water fish and fish oil has been reported to protect against obesity. Increased attention has been focused on dietary ω-3 polyunsaturated fatty acids (PUFAs), mainly EPA (20:5; ω-3) and DHA (22:6; ω-3), which have been reported to protect against the development of obesity and reduce body fat in humans [2, 3]. Cold water fish and fish oils are the most direct source of EPA and DHA to the body. Consumption of stearidonic acid (SDA; 18:4; ω-3), the metabolic intermediate between ALA and EPA (Fig. 1), was found to result in significant EPA enrichment due to bypassing the Δ6 desaturase enzyme in ω-3 fatty acid metabolism [7]. Dietary SDA increased red blood cell EPA by approximately 17%, whereas the efficiency of ALA was about 0.1% [8]
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