(P06-104-20) Eicosapentaenoic Acid-Induced Inhibition of Metabolic Inflammation Is Associated with Preserved Mitochondrial Function and Insulin Sensitivity in Human Primary Myotubes

Objectives: The aim of this study was to investigate whether metabolic inflammation in skeletal muscle may be prevented by eicosapentaenoic acid (EPA) and if this is associated with an improvement in markers of mitochondrial function and insulin sensitivity.

Methods: Human primary myotubes were treated for 24 hours with palmitic acid (PA, 500 µM) in hyperglycaemic conditions (13 mM glucose), referred to as nutrient overload, in the presence or absence of EPA (100 µM). After the treatments, the expression of peroxisome proliferator-activated receptor γ coactivator 1-alpha (PGC1α) and interleukin-6 (IL-6) was assessed by q-PCR. Western blot was used to asses the abundance of the inhibitor of nuclear factor kappa-B (IKBα), mitochondrial electron transport chain complex proteins, the phosphorylation of AKT (Ser473) and AKT substrate 160 (AS 160) (Thr642) in response to insulin, the activation of 5'-AMP-activated protein kinase (AMPK) and the inhibition of acetyl-CoA carboxylase (ACC). Mitochondrial dynamics was assessed by immunocytochemistry.

Results: Nutrient excess activated the proinflammatory nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB) signalling as indicated by the upregulation of IL-6 mRNA (p< 0.001) and a tendency to decrease in IKBα (p=0.0654), tended to downregulate PGC1α (p=0.0589) and promoted mitochondrial fragmentation (p< 0.001), all of which were counteracted by EPA. Furthermore, EPA  induced complex III-core protein 2 (p< 0.05) relative to control cells, an effect that was absent in the myotubes exposed only to PA and hyperglycaemia. EPA, when administrated in combination with PA and hyperglycaemia, induced the phosphorylation of AMPK (p< 0.05) and its downstream target ACC (p< 0.05) relative to cells exposed to nutrient overload alone. Finally, while fuel surplus impaired insulin-induced phosphorylation of AKT (p< 0.01) and AS160 (p< 0.05), these effects were prevented by EPA.

Conclusions: EPA inhibited NFkB signalling which was associated with an attenuation of the deleterious effects of PA and hyperglycaemia on markers of mitochondrial function and insulin sensitivity. Thus, EPA may represent a valuable nutritional tool to preserve skeletal muscle mitochondrial function and metabolic health during periods of nutrient overload.

Funding Sources: CSIRO's Precision Health Future Science Platform (FSP)