The dose–response effects of arachidonic acid on primary human skeletal myoblasts and myotubes
Background Cellular inflammatory response, mediated by arachidonic acid (AA) and cyclooxygenase, is a highly regulated process that leads to the repair of damaged tissue. Recent studies on murine C2C12 cells have demonstrated that AA supplementation leads to myotube hypertrophy. However, AA has not...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2023-12-01
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| Series: | Journal of the International Society of Sports Nutrition |
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| Online Access: | http://dx.doi.org/10.1080/15502783.2022.2164209 |
| _version_ | 1797679042592243712 |
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| author | Brandon M. Roberts Alexander L. Kolb Alyssa V. Geddis Marshall A. Naimo Ronald W. Matheny |
| author_facet | Brandon M. Roberts Alexander L. Kolb Alyssa V. Geddis Marshall A. Naimo Ronald W. Matheny |
| author_sort | Brandon M. Roberts |
| collection | DOAJ |
| description | Background Cellular inflammatory response, mediated by arachidonic acid (AA) and cyclooxygenase, is a highly regulated process that leads to the repair of damaged tissue. Recent studies on murine C2C12 cells have demonstrated that AA supplementation leads to myotube hypertrophy. However, AA has not been tested on primary human muscle cells. Therefore, the purpose of this study was to determine whether AA supplementation has similar effects on human muscle cells. Methods Proliferating and differentiating human myoblasts were exposed to AA in a dose-dependent manner (50–0.80 µM) for 48 (myoblasts) or 72 (myotubes) hours. Cell viability was tested using a 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay and cell counting; myotube area was determined by immunocytochemistry and confocal microscopy; and anabolic signaling pathways were evaluated by western blot and RT-PCR. Results Our data show that the treatment of primary human myoblasts treated with 50 µM and 25 µM of AA led to the release of PGE2 and PGF2α at levels higher than those of control-treated cells (p < 0.001 for all concentrations). Additionally, 50 µM and 25 µM of AA suppressed myoblast proliferation, myotube area, and myotube fusion. Anabolic signaling indicated reductions in total and phosphorylated TSC2, AKT, S6, and 4EBP1 in myoblasts at 50 µM of AA (p < 0.01 for all), but not in myotubes. These changes were not affected by COX-2 inhibition with celecoxib. Conclusion Together, our data demonstrate that high concentrations of AA inhibit myoblast proliferation, myotube fusion, and myotube hypertrophy, thus revealing potential deleterious effects of AA on human skeletal muscle cell health and viability. |
| first_indexed | 2024-03-11T23:07:42Z |
| format | Article |
| id | doaj.art-b2ba846b26af4b1fb47ebc4149c72dd2 |
| institution | Directory Open Access Journal |
| issn | 1550-2783 |
| language | English |
| last_indexed | 2024-03-11T23:07:42Z |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Journal of the International Society of Sports Nutrition |
| spelling | doaj.art-b2ba846b26af4b1fb47ebc4149c72dd22023-09-21T12:43:10ZengTaylor & Francis GroupJournal of the International Society of Sports Nutrition1550-27832023-12-0120110.1080/15502783.2022.21642092164209The dose–response effects of arachidonic acid on primary human skeletal myoblasts and myotubesBrandon M. Roberts0Alexander L. Kolb1Alyssa V. Geddis2Marshall A. Naimo3Ronald W. Matheny4US Army Research Institute of Environmental MedicineUS Army Research Institute of Environmental MedicineUS Army Research Institute of Environmental MedicineUS Army Research Institute of Environmental MedicineUS Army Research Institute of Environmental MedicineBackground Cellular inflammatory response, mediated by arachidonic acid (AA) and cyclooxygenase, is a highly regulated process that leads to the repair of damaged tissue. Recent studies on murine C2C12 cells have demonstrated that AA supplementation leads to myotube hypertrophy. However, AA has not been tested on primary human muscle cells. Therefore, the purpose of this study was to determine whether AA supplementation has similar effects on human muscle cells. Methods Proliferating and differentiating human myoblasts were exposed to AA in a dose-dependent manner (50–0.80 µM) for 48 (myoblasts) or 72 (myotubes) hours. Cell viability was tested using a 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay and cell counting; myotube area was determined by immunocytochemistry and confocal microscopy; and anabolic signaling pathways were evaluated by western blot and RT-PCR. Results Our data show that the treatment of primary human myoblasts treated with 50 µM and 25 µM of AA led to the release of PGE2 and PGF2α at levels higher than those of control-treated cells (p < 0.001 for all concentrations). Additionally, 50 µM and 25 µM of AA suppressed myoblast proliferation, myotube area, and myotube fusion. Anabolic signaling indicated reductions in total and phosphorylated TSC2, AKT, S6, and 4EBP1 in myoblasts at 50 µM of AA (p < 0.01 for all), but not in myotubes. These changes were not affected by COX-2 inhibition with celecoxib. Conclusion Together, our data demonstrate that high concentrations of AA inhibit myoblast proliferation, myotube fusion, and myotube hypertrophy, thus revealing potential deleterious effects of AA on human skeletal muscle cell health and viability.http://dx.doi.org/10.1080/15502783.2022.2164209cyclooxygenasemyogenesisanabolic signalingskeletal muscle |
| spellingShingle | Brandon M. Roberts Alexander L. Kolb Alyssa V. Geddis Marshall A. Naimo Ronald W. Matheny The dose–response effects of arachidonic acid on primary human skeletal myoblasts and myotubes Journal of the International Society of Sports Nutrition cyclooxygenase myogenesis anabolic signaling skeletal muscle |
| title | The dose–response effects of arachidonic acid on primary human skeletal myoblasts and myotubes |
| title_full | The dose–response effects of arachidonic acid on primary human skeletal myoblasts and myotubes |
| title_fullStr | The dose–response effects of arachidonic acid on primary human skeletal myoblasts and myotubes |
| title_full_unstemmed | The dose–response effects of arachidonic acid on primary human skeletal myoblasts and myotubes |
| title_short | The dose–response effects of arachidonic acid on primary human skeletal myoblasts and myotubes |
| title_sort | dose response effects of arachidonic acid on primary human skeletal myoblasts and myotubes |
| topic | cyclooxygenase myogenesis anabolic signaling skeletal muscle |
| url | http://dx.doi.org/10.1080/15502783.2022.2164209 |
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