Exploring the Role of Oleic Acid in Muscle Cell Differentiation: Mechanisms and Implications for Myogenesis and Metabolic Regulation in C2C12 Myoblasts

<b>Background/Objectives</b>: Myogenesis, the process by which myoblasts differentiate into multinucleated muscle fibers, is tightly regulated by transcription factors, signaling pathways, and metabolic cues. Among these, fatty acids have emerged as key regulators beyond their traditiona...

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Main Authors: Francesco Vari, Elisa Bisconti, Ilaria Serra, Eleonora Stanca, Marzia Friuli, Daniele Vergara, Anna Maria Giudetti
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Biomedicines
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Online Access:https://www.mdpi.com/2227-9059/13/7/1568
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author Francesco Vari
Elisa Bisconti
Ilaria Serra
Eleonora Stanca
Marzia Friuli
Daniele Vergara
Anna Maria Giudetti
author_facet Francesco Vari
Elisa Bisconti
Ilaria Serra
Eleonora Stanca
Marzia Friuli
Daniele Vergara
Anna Maria Giudetti
author_sort Francesco Vari
collection DOAJ
description <b>Background/Objectives</b>: Myogenesis, the process by which myoblasts differentiate into multinucleated muscle fibers, is tightly regulated by transcription factors, signaling pathways, and metabolic cues. Among these, fatty acids have emerged as key regulators beyond their traditional role as energy substrates. Oleic acid, a monounsaturated fatty acid, has been shown to modulate muscle differentiation, potentially influencing myogenic pathways. This study examines the role of oleic acid in promoting C2C12 myoblast differentiation and its associated molecular mechanisms, comparing it to standard horse serum (HS)-based differentiation protocols. <b>Methods</b>: C2C12 murine myoblasts were cultured under proliferative conditions and differentiated using DMEM supplemented with either 2% HS or oleic acid (C18:1, n-9). The molecular signaling pathway was evaluated by measuring the expression of p38 MAPK, β-catenin, GLUT4, and NDRG1. <b>Results</b>: Oleic acid promoted the differentiation of C2C12 cells, as evidenced by a progressively elongated morphology, as well as the induction of muscle-specific myogenin, myosin heavy chain (MHC), and <i>MyoD</i>. Moreover, oleic acid reduced the expression of Atrogin-1 and <i>MuRF1</i> ubiquitin E3 ligase. BODIPY staining revealed the enhanced accumulation of lipid droplets in oleic acid-treated cells. The Western blot analysis demonstrated robust activation of p38 MAPK and β-catenin pathways in response to oleic acid, compared with HS. Additionally, oleic acid upregulated GLUT4 expression and increased the phosphorylation of insulin receptor and NDRG1, indicating an enhanced glucose uptake capacity. <b>Conclusions</b>: These findings demonstrate that oleic acid promotes C2C12 myoblast differentiation and improves glucose uptake via GLUT4. Oleic acid emerges as a promising metabolic regulator of myogenesis, offering potential therapeutic applications for muscle regeneration in muscle-related pathologies.
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spelling doaj-art-3b2b59ba0fa94aeb8e2abfb1ee450bcc2025-08-20T03:36:02ZengMDPI AGBiomedicines2227-90592025-06-01137156810.3390/biomedicines13071568Exploring the Role of Oleic Acid in Muscle Cell Differentiation: Mechanisms and Implications for Myogenesis and Metabolic Regulation in C2C12 MyoblastsFrancesco Vari0Elisa Bisconti1Ilaria Serra2Eleonora Stanca3Marzia Friuli4Daniele Vergara5Anna Maria Giudetti6Department of Physiology and Pharmacology, Sapienza University of Rome, 00185 Rome, ItalyDepartment of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Lecce, ItalyDepartment of Physiology and Pharmacology, Sapienza University of Rome, 00185 Rome, ItalyDepartment of Experimental Medicine, University of Salento, 73100 Lecce, ItalyDepartment of Physiology and Pharmacology, Sapienza University of Rome, 00185 Rome, ItalyDepartment of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Lecce, ItalyDepartment of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Lecce, Italy<b>Background/Objectives</b>: Myogenesis, the process by which myoblasts differentiate into multinucleated muscle fibers, is tightly regulated by transcription factors, signaling pathways, and metabolic cues. Among these, fatty acids have emerged as key regulators beyond their traditional role as energy substrates. Oleic acid, a monounsaturated fatty acid, has been shown to modulate muscle differentiation, potentially influencing myogenic pathways. This study examines the role of oleic acid in promoting C2C12 myoblast differentiation and its associated molecular mechanisms, comparing it to standard horse serum (HS)-based differentiation protocols. <b>Methods</b>: C2C12 murine myoblasts were cultured under proliferative conditions and differentiated using DMEM supplemented with either 2% HS or oleic acid (C18:1, n-9). The molecular signaling pathway was evaluated by measuring the expression of p38 MAPK, β-catenin, GLUT4, and NDRG1. <b>Results</b>: Oleic acid promoted the differentiation of C2C12 cells, as evidenced by a progressively elongated morphology, as well as the induction of muscle-specific myogenin, myosin heavy chain (MHC), and <i>MyoD</i>. Moreover, oleic acid reduced the expression of Atrogin-1 and <i>MuRF1</i> ubiquitin E3 ligase. BODIPY staining revealed the enhanced accumulation of lipid droplets in oleic acid-treated cells. The Western blot analysis demonstrated robust activation of p38 MAPK and β-catenin pathways in response to oleic acid, compared with HS. Additionally, oleic acid upregulated GLUT4 expression and increased the phosphorylation of insulin receptor and NDRG1, indicating an enhanced glucose uptake capacity. <b>Conclusions</b>: These findings demonstrate that oleic acid promotes C2C12 myoblast differentiation and improves glucose uptake via GLUT4. Oleic acid emerges as a promising metabolic regulator of myogenesis, offering potential therapeutic applications for muscle regeneration in muscle-related pathologies.https://www.mdpi.com/2227-9059/13/7/1568C2C12 myoblast differentiationglucose uptakelipid dropletsoleic acidskeletal muscle
spellingShingle Francesco Vari
Elisa Bisconti
Ilaria Serra
Eleonora Stanca
Marzia Friuli
Daniele Vergara
Anna Maria Giudetti
Exploring the Role of Oleic Acid in Muscle Cell Differentiation: Mechanisms and Implications for Myogenesis and Metabolic Regulation in C2C12 Myoblasts
Biomedicines
C2C12 myoblast differentiation
glucose uptake
lipid droplets
oleic acid
skeletal muscle
title Exploring the Role of Oleic Acid in Muscle Cell Differentiation: Mechanisms and Implications for Myogenesis and Metabolic Regulation in C2C12 Myoblasts
title_full Exploring the Role of Oleic Acid in Muscle Cell Differentiation: Mechanisms and Implications for Myogenesis and Metabolic Regulation in C2C12 Myoblasts
title_fullStr Exploring the Role of Oleic Acid in Muscle Cell Differentiation: Mechanisms and Implications for Myogenesis and Metabolic Regulation in C2C12 Myoblasts
title_full_unstemmed Exploring the Role of Oleic Acid in Muscle Cell Differentiation: Mechanisms and Implications for Myogenesis and Metabolic Regulation in C2C12 Myoblasts
title_short Exploring the Role of Oleic Acid in Muscle Cell Differentiation: Mechanisms and Implications for Myogenesis and Metabolic Regulation in C2C12 Myoblasts
title_sort exploring the role of oleic acid in muscle cell differentiation mechanisms and implications for myogenesis and metabolic regulation in c2c12 myoblasts
topic C2C12 myoblast differentiation
glucose uptake
lipid droplets
oleic acid
skeletal muscle
url https://www.mdpi.com/2227-9059/13/7/1568
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