Effects of eicosapentaenoic acid and docosahexaenoic acid on C2C12 cell adipogenesis and inhibition of myotube formation

Abstract

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) modulate cellular metabolic functions and gene expression. This study investigated the impacts of EPA and DHA on gene expression and morphological changes during adipogenic inducement in C2C12 myoblasts. Cells were cultured and treated with differentiation medium with and without 50 μM EPA and DHA. Cells treated with fatty acids had noticeable lipid droplets, but no formation of myotubes compared to control group cells. The expression levels of key genes relevant to adipogenesis and inflammation were significantly higher (P < 0.05) in cells treated with fatty acids. Genes associated with myogenesis and mitochondrial biosynthesis and function had lower (P < 0.05) expression with fatty acids supplementation. Moreover, fatty acid treatment reduced (P < 0.05) oxygen consumption rate in the differentiated cells. This suggested blocking myotube formation through supplementation with EPA and DHA drove myoblasts to enter the quiescent state and enabled adipogenic trans-differentiation of the myoblasts. Data also suggested that overdosage of EPA and DHA during gestation may drive fetal mesenchymal stem cell differentiation to the fate of adipogenesis and have a long-term effect on childhood obesity.

Divergent Regulation of Myotube Formation and Gene Expression by E2 and EPA during In-Vitro Differentiation of C2C12 Myoblasts

“…Our finding that n-3PUFA (i.e., EPA) inhibits myogenesis is in agreement with accumulating evidence showing the negative effect of n-3PUFA on myotube formation in-vitro [17,19,22]. These studies highlight the potential negative effects of n-3PUFA consumption during pregnancy when de novo myogenesis is initiated at the embryonic and neonatal stages because disruption to myogenesis at this time may lead to postnatal muscle deficiencies in mass and function [50].…”

“…In contrast to our original research hypothesis, cotreatment of E2 and n-3PUFA did not have a synergistic effect on in-vitro myogenesis, raising the possibility that dietary supplements of fish oil (i.e., n-3PUFA) may interfere with the positive affect estrogen has on muscle regeneration in women on HRT. Recently, Ghnaimawi et al [17] showed that treatment of differentiating C2C12 myoblasts with 50 µM EPA and DHA for four days increased adipogenesis and inflammatory-related genes and reduced tube formation. We did not examine adipose cell formation in our cultures following treatment with EPA, but were able to identify that IL6 and adipose tissue related genes were uniquely expressed in cells treated with EPA (Tables S3 and S4).…”

“…Fish oil contains n-3PUFA that are known to improve muscle strength, particularly in older women [10,11]. However, while some studies find supportive effects of E2 or n-3PUFA on myoblast differentiation in-vitro [12][13][14][15][16], others have shown an inhibition [17][18][19][20][21][22].…”

“…In the present study we tested the hypothesis that E2 and n-3PUFA would induce a positive and synergistic effect on myogenesis in-vitro. From the numerous n-3PUFA investigated to date, eicosapentaenoic acid (EPA) has been found to have the most significant effect regulating myogenesis in-vitro [14,16,17,22]. Hence, we examined myogenesis fate in mouse C2C12 myoblasts treated with E2 and EPA independently or in combination, for 0-24 h or 0-120 h from induction of differentiation, with a focus on morphological changes and differential expression of key genes involved in muscle differentiation and muscle function pathways.…”

Integrated metabolomic and transcriptomic analyses identify critical genes in eicosapentaenoic acid biosynthesis and metabolism in the sea urchin Strongylocentrotus intermedius

“…www.nature.com/scientificreports www.nature.com/scientificreports/ Very long-chain 3-oxoacyl-CoA reductase (Hsd17b12) catalyzes the second of the four reactions of the long-chain fatty acids elongation cycle, and this enzyme has 3-ketoacyl-CoA reductase activity, reducing 3-ketoacyl-CoA to 3-hydroxyacyl-CoA, within each cycle of fatty acid elongation. It may participate in the production of PUFAs of different chain lengths that are involved in multiple biological processes as precursors of membrane lipids and lipid mediators 34 , especially like EPA, which may potentially drive myoblasts to enter the quiescent state and enable adipogenic trans-differentiation of the myoblasts and impairs brown adipogenesis and thermogenic capacity 35,36 .…”