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Cited by 112Fatty Acid Research and HealthAdipose Tissue and MetabolismMitochondrial Function and Pathology

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Conjugated linoleic acid or omega 3 fatty acids increase mitochondrial biosynthesis and metabolism in skeletal muscle cells

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Vaughan, Roger A, Garcia-Smith, Randi, Bisoffi, Marco, Conn, Carole A, Trujillo, Kristina A·2012·Lipids in Health and Disease
DOI 10.1186/1476-511x-11-142PMC PMC3515476
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Abstract

BACKGROUND: Polyunsaturated fatty acids are popular dietary supplements advertised to contribute to weight loss by increasing fat metabolism in liver, but the effects on overall muscle metabolism are less established. We evaluated the effects of conjugated linoleic acid (CLA) or combination omega 3 on metabolic characteristics in muscle cells. METHODS: Human rhabdomyosarcoma cells were treated with either DMSO control, or CLA or combination omega 3 for 24 or 48 hours. RNA was determined using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Mitochondrial content was determined using flow cytometry and immunohistochemistry. Metabolism was quantified by measuring extracellular acidification and oxygen consumption rates. RESULTS: Omega 3 significantly induced metabolic genes as well as oxidative metabolism (oxygen consumption), glycolytic capacity (extracellular acidification), and metabolic rate compared with control. Both treatments significantly increased mitochondrial content. CONCLUSION: Omega 3 fatty acids appear to enhance glycolytic, oxidative, and total metabolism. Moreover, both omega 3 and CLA treatment significantly increase mitochondrial content compared with control.

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Figure 1 Polyunsaturated fatty acids modify glycolytic metabolism. A- Extracellular acidification rate (ECAR) of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 hours. B- Baseline ECAR following treatment described above. C- Peak ECAR following addition of oligomycin, an inhibitor of oxidative phosphorylation. D- Peak ECAR following addition of carbonyl cyanide p-[trifluoromethoxy]-phenyl-hydrazone (FCCP), a mitochondrial uncoupling agent, in addition to previously added oligomycin. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control.
Figure 1 Polyunsaturated fatty acids modify glycolytic metabolism. A- Extracellular acidification rate (ECAR) of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 hours. B- Baseline ECAR following treatment described above. C- Peak ECAR following addition of oligomycin, an inhibitor of oxidative phosphorylation. D- Peak ECAR following addition of carbonyl cyanide p-[trifluoromethoxy]-phenyl-hydrazone (FCCP), a mitochondrial uncoupling agent, in addition to previously added oligomycin. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control.CC BY · PubMed Central
Figure 2 Polyunsaturated fatty acids modify oxidative metabolism. A- Oxygen consumption rate (OCR) of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 hours. B- Baseline OCR following treatment described above. C- OCR following addition of oligomycin, an inhibitor of oxidative phosphorylation. D- Peak OCR following addition of carbonyl cyanide p-[trifluoromethoxy]-phenyl-hydrazone (FCCP), a mitochondrial uncoupling agent, in addition to previously added oligomycin. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control. † indicates p < 0.01 (significantly less than control).
Figure 2 Polyunsaturated fatty acids modify oxidative metabolism. A- Oxygen consumption rate (OCR) of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 hours. B- Baseline OCR following treatment described above. C- OCR following addition of oligomycin, an inhibitor of oxidative phosphorylation. D- Peak OCR following addition of carbonyl cyanide p-[trifluoromethoxy]-phenyl-hydrazone (FCCP), a mitochondrial uncoupling agent, in addition to previously added oligomycin. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control. † indicates p < 0.01 (significantly less than control).CC BY · PubMed Central
Figure 3 Polyunsaturated fatty acids modify oxidative reliance OCR:ECAR. A- Ratio of oxygen consumption rate (OCR) to extracellular acidification rate (ECAR) of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 hours. B- Relative glycolytic reliance at baseline OCR:ECAR following treatment described above with control normalized to value of 1. C- Relative glycolytic reliance from OCR:ECAR following addition of oligomycin (peak glycolysis), an inhibitor of oxidative phosphorylation. D- Relative glycolytic reliance with control = 1 from OCR:ECAR following addition of carbonyl cyanide p-[trifluoromethoxy]-phenyl-hydrazone (FCCP), a mitochondrial uncoupling agent, (peak oxidation) in addition to previously added oligomycin. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control. † CLA had increased OCR:ECAR but had lower total individual OCR and ECAR compared with control.
Figure 3 Polyunsaturated fatty acids modify oxidative reliance OCR:ECAR. A- Ratio of oxygen consumption rate (OCR) to extracellular acidification rate (ECAR) of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 hours. B- Relative glycolytic reliance at baseline OCR:ECAR following treatment described above with control normalized to value of 1. C- Relative glycolytic reliance from OCR:ECAR following addition of oligomycin (peak glycolysis), an inhibitor of oxidative phosphorylation. D- Relative glycolytic reliance with control = 1 from OCR:ECAR following addition of carbonyl cyanide p-[trifluoromethoxy]-phenyl-hydrazone (FCCP), a mitochondrial uncoupling agent, (peak oxidation) in addition to previously added oligomycin. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control. † CLA had increased OCR:ECAR but had lower total individual OCR and ECAR compared with control.CC BY · PubMed Central
Figure 4 Relative metabolic rate represented by oxygen consumption rate (OCR) versus extracellular acidification rate (ECAR) of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration) or combination omega 3 (Ω3) at 25 μM or 50 μM for 24 hours. NOTES: * indicates p < 0.05. Combination omega 3 fatty acids increase basal metabolic rate.
Figure 4 Relative metabolic rate represented by oxygen consumption rate (OCR) versus extracellular acidification rate (ECAR) of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration) or combination omega 3 (Ω3) at 25 μM or 50 μM for 24 hours. NOTES: * indicates p < 0.05. Combination omega 3 fatty acids increase basal metabolic rate.CC BY · PubMed Central
Figure 5 Relative expression of PGC-1α (top row), GLUT4 (middle row) and Irisin (bottom row) following treatment with DMSO control (final concentration 0.1%), combination omega 3 at 50 μM, or CLA at 25 μM or 50 μM for either 24 hours (left), 48 hours with repeated treatment at 24 hours (center), or single treatment for 48 hours (right) with control = 1. A- PGC-1α expression following treatment for 24 hours. B- PGC-1α expression following repeated treatment for 48 hours. C- PGC-1α expression following treatment for 48 hours. D- GLUT4 expression following treatment for 24 hours. E- GLUT4 expression following repeated treatment for 48 hours. F- GLUT4 expression following treatment for 48 hours. G- Irisin expression following treatment for 24 hours. H- Irisin expression following repeated treatment for 48 hours. I- Irisin expression following treatment for 48 hours. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control. Polyunsaturated fatty acids modify metabolic gene expression.
Figure 5 Relative expression of PGC-1α (top row), GLUT4 (middle row) and Irisin (bottom row) following treatment with DMSO control (final concentration 0.1%), combination omega 3 at 50 μM, or CLA at 25 μM or 50 μM for either 24 hours (left), 48 hours with repeated treatment at 24 hours (center), or single treatment for 48 hours (right) with control = 1. A- PGC-1α expression following treatment for 24 hours. B- PGC-1α expression following repeated treatment for 48 hours. C- PGC-1α expression following treatment for 48 hours. D- GLUT4 expression following treatment for 24 hours. E- GLUT4 expression following repeated treatment for 48 hours. F- GLUT4 expression following treatment for 48 hours. G- Irisin expression following treatment for 24 hours. H- Irisin expression following repeated treatment for 48 hours. I- Irisin expression following treatment for 48 hours. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control. Polyunsaturated fatty acids modify metabolic gene expression.CC BY · PubMed Central
Figure 6 Polyunsaturated fatty acids increase mitochondrial content. A-Flow cytometry using mitochondrial staining of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 hours. B- Flow cytometry using mitochondrial staining following similar treatment described above for 48 hours with repeat treatment at 24 hours. C- Flow cytometry using mitochondrial staining following a single treatment described above for 48 hours. D- Immunohistochemistry of cells treated as described in Figure 4A and stained with Mitotracker (green) and DAPI (blue) with 0.1% DMSO control (left), Ω3 50 μM (middle) and CLA 50 μM (right). Red line indicates 50 μm and the red arrow indicates mitochondrial networking. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control.
Figure 6 Polyunsaturated fatty acids increase mitochondrial content. A-Flow cytometry using mitochondrial staining of rhabdomyosarcoma cells treated with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 hours. B- Flow cytometry using mitochondrial staining following similar treatment described above for 48 hours with repeat treatment at 24 hours. C- Flow cytometry using mitochondrial staining following a single treatment described above for 48 hours. D- Immunohistochemistry of cells treated as described in Figure 4A and stained with Mitotracker (green) and DAPI (blue) with 0.1% DMSO control (left), Ω3 50 μM (middle) and CLA 50 μM (right). Red line indicates 50 μm and the red arrow indicates mitochondrial networking. NOTES: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared with control.CC BY · PubMed Central
Figure 7 Cell viability. Measured by group mean log fluorescence from WST-1 end-point viability and proliferation assay following treatment of rhabdomyosarcoma cells with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 (A) or 48 hours (B).
Figure 7 Cell viability. Measured by group mean log fluorescence from WST-1 end-point viability and proliferation assay following treatment of rhabdomyosarcoma cells with either DMSO control (0.1% final concentration), combination omega 3 (Ω3) at 25 μM or 50 μM, or CLA at 25 μM or 50 μM for 24 (A) or 48 hours (B).CC BY · PubMed Central

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  1. Duda MK, Xu WH, Tintinu A, O'Shea KM, Stanley WC: Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) Supplementation, but not alpha-Linolenic Acid, Elevates Plasma Adiponectin Concentration and Prevents Pressure Overload Induced Ventricular Dysfunction and Remodeling. Circulation. 2008, 118: S542-S542.

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Metadata

Authors
Vaughan, R. A., Garcia-Smith, R., Bisoffi, M., Conn, C. A., & Trujillo, K. A.
Affiliations
University of New Mexico (US)
Corresponding author
Vaughan, Roger A
ORCID
Venue
Lipids in Health and Disease
Publisher
Springer Science and Business Media LLC
Published
2012-10-30
Volume
11
Issue
1
Pages
142-142
ISSN
1476-511X (ISSN-L)
DOI
10.1186/1476-511x-11-142
PMC
PMC3515476
OpenAlex
W2117258334
Cited by
112
References
48
Citations by year
2026: 1 · 2025: 7 · 2024: 4 · 2023: 9 · 2022: 8 · 2021: 6 · 2020: 11 · 2019: 5 · 2018: 9 · 2017: 10 · 2016: 13 · 2015: 12 · 2014: 10 · 2013: 6 · 2012: 1
FWCI
8.66
Type
journal article
Language
en
Open access
Licence
Locations
  • Lipids in Health and Disease (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Topics
  • Health Sciences › Nursing › Nutrition and Dietetics › Fatty Acid Research and Health
  • Health Sciences › Medicine › Physiology (medical) › Adipose Tissue and Metabolism
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
Keywords
conjugated linoleic acid, omega-3 fatty acids, mitochondrial biogenesis, skeletal muscle metabolism, rhabdomyosarcoma cells, glycolysis, oxidative metabolism, extracellular acidification, oxygen consumption, polyunsaturated fatty acids, dietary supplements
MeSH terms
Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha, Base Sequence, DNA, Fibronectins, Glycolysis, Heat-Shock Proteins, Humans, Mitochondria, Muscle, Oxygen Consumption, Transcription Factors, Tumor Cells, Cultured, Fatty Acids, Omega-3, Gene Expression, Muscle Fibers, Skeletal, Dietary Supplements, Linoleic Acids, Conjugated, Glucose Transporter Type 4

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