Methylation analysis in fatty-acid-related genes reveals their plasticity associated with conjugated linoleic acid and calcium supplementation in adult mice
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DNA methylation is one of the most extensively studied mechanisms within epigenetics, and it is suggested that diet-induced changes in methylation status could be involved in energy metabolism regulation. Conjugated linoleic acid (CLA) and calcium supplementation counteract body weight gain, particularly under a high-fat (HF) diet, in adult mice. The aim was to determine whether the modulation of DNA methylation pattern in target genes and tissues could be an underlying mechanism of action.
Mice (C57BL/6J) were divided into five groups according to diet and treatment: normal fat as the control group (12 % kJ content as fat), HF group (43 % kJ content as fat), HF + CLA (6 mg CLA/day), HF + calcium (12 g/kg of calcium) and HF with both compounds. Gene expression and methylation degree of CpG sites in promoter sequences of genes involved in fatty acid metabolism, including adiponectin (Adipoq), stearoyl-CoA desaturase (Scd1) and fatty acid synthase (Fasn), were determined by bisulphite sequencing in liver and epididymal white adipose tissue.
Results showed that the methylation profile of promoters was significantly altered by dietary supplementation in a gene- and tissue-specific manner, whereas only slight changes were observed in the HF group. Furthermore, changes in specific CpG sites were also associated with an overall healthier metabolic profile, in particular for calcium-receiving groups.
Both CLA and calcium were able to modify the methylation pattern of genes involved in energy balance in adulthood, which opens a novel area for increasing efficiency in body weight management strategies.
KeywordsCalcium supplementation Conjugated linoleic acid DNA methylation Fatty acid synthase Stearoyl-CoA desaturase Adiponectin
We thank Pilar Parra for her assistance in the first steps of experimental design, animal handling and sample collection, and Sarah Laraichi (Laboratory of Calorimetry and Materials, Faculty of Sciences, Abdelmalek Essaâdi University, 93030 Tétouan, Morocco) for her help with the animal care and sample collection during her stay in our laboratory. This work was supported by the Grants AGL2012-33692 and Fundación Ramón Areces (XVI Concurso Nacional, ref. Nutriepigenética y adiposidad). Our group receives financial support from Instituto de Salud Carlos III and Centro de Investigación Biomédica en Red Fisiopatología de la Obesidad y Nutrición (CIBERobn). Authors belong to the Nutrigenomics group, awarded as “Group of Excellence” by the CAIB and supported by the “Direcció General d’Universitats, Recerca i Transferència del Coneixement” of the Regional Government (CAIB) and FEDER funds (EU). A.C. has been supported by a PhD fellowship by the Conselleria d’Educació, Cultura i Universitats, Govern de les Illes Balears, as part of a programme co-financed by the European Social Fund.
Compliance with ethical standards
Conflict of interest
On behalf of all authors, the corresponding author states that there is no conflict of interest.
All applicable international, national and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. The animal protocol followed in this study was reviewed and approved by the Bioethical Committee of the University of the Balearic Islands (approval 13 February 2006).
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