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Alterations in oxidative stress status modulate terminal differentiation in Atlantic salmon adipocytes cultivated in media rich in n-3 fatty acids

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Comparative Biochemistry and Physiology - Part B: Biochemistry & Molecular Biology ; Volume 156. p. 309–318. 2010

Todorcevic, Marijana; Skugor, Stanko; Ruyter, Bente

Regulation of oxidative stress (OS) in adipocytes is an important mediator of their development and dysfunction. Highly unsaturated fatty acids (HUFAs) play essential roles in marine fish, where they have anti-lipogenic effects, but they are prone to peroxidation. The aim of this study was to investigate how the effects of HUFAs in fish adipocytes are modulated by changes in their intracellular redox status. Adipocytes from Atlantic salmon were cultivated on HUFA-rich media and treated with buthionine sulfoximine (BSO), which is known to deplete stores of the antioxidant glutathione (GSH) thus increasing OS, and alpha-tocopherol (alpha-TOC), which protects from OS. Gene expression was assessed by qPCR. In addition, phospholipid composition, total fatty acid (FA) composition, TBARS, the activities of pro-apoptotic caspase 3 (CASP3) and antioxidant superoxide dismutase (SOD) were determined. BSO treatment decreased the expression of genes encoding GSH-based antioxidant enzymes glutathione peroxidase (GPX) 2 and GPX3. Consequently, depletion of GSH resulted in the highest level of peroxidation products TBARS despite the increased activity of SOD in this group. Significant reduction of TBARS was achieved by alpha-TOC. Further, in comparison to two alpha-TOC supplemented groups, GSH-depleted cells accumulated less fat and their gene expression profile of adipogenic markers was lower. The formation of large intracellular vesicles was prominent in the control and BSO groups while reduction of OS by alpha-TOC coincided with the increased gene expression of the activating transcription factor 6 (ATF6), a transducer of the endoplasmic reticulum (ER)-stress response. CASP3 assay showed no difference between groups; however, depletion of GSH resulted in the increased gene expression of several apoptotic markers. Up-regulation of the apoptosis inducible factor (AIF) implied higher probability of CASP3-independent apoptosis in cells under increased OS. In conclusion, the study provides several lines of evidence in favour of anti-adipogenic effects of OS in a cold blooded vertebrate. (C) 2010 Elsevier Inc. All rights reserved.

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