The Effects of Creatine Monohydrate and/or Whey Protein on the Muscle Protein Synthesis and Anabolic Signaling Responses in Non-Stressed C<sub>2</sub>C<sub>12</sub> Murine Myotubes
<b>Background/Objectives</b>: Creatine monohydrate (CRE) is a popular nutritional supplement that increases lean/muscle mass accretion. Although data regarding CRE and its effects on muscle protein synthesis are mixed, we hypothesized that CRE may potentiate/extend the anabolic response...
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2025-05-01
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| author | Nicholas J. Kontos Joshua S. Godwin Anthony Agyin-Birikorang Darren G. Candow Christopher M. Lockwood Michael D. Roberts Christopher B. Mobley |
| author_facet | Nicholas J. Kontos Joshua S. Godwin Anthony Agyin-Birikorang Darren G. Candow Christopher M. Lockwood Michael D. Roberts Christopher B. Mobley |
| author_sort | Nicholas J. Kontos |
| collection | DOAJ |
| description | <b>Background/Objectives</b>: Creatine monohydrate (CRE) is a popular nutritional supplement that increases lean/muscle mass accretion. Although data regarding CRE and its effects on muscle protein synthesis are mixed, we hypothesized that CRE may potentiate/extend the anabolic response to essential amino acids given that CRE acts as a high-energy phosphate buffer to potentially amplify anabolic signaling. Therefore, we used an in vitro model to determine whether CRE synergistically enhances myotube protein synthesis and the anabolic signaling responses to EAA-rich whey protein (WP). <b>Methods</b>: C<sub>2</sub>C<sub>12</sub> murine myotubes were treated with control media containing PBS (CTL), WP serum (5 mg/mL), CRE (10 mM), or WP + CRE. Myotubes were collected following 1, 4, and 24 h treatments (n = 6 replicates per treatment and time point) and assayed for relative creatine levels, myotube protein synthesis levels, and phosphorylation markers. <b>Results</b>: Cellular creatine levels were greater in CRE and WP + CRE versus CTL and WP at all treatment time points (<i>p</i> < 0.05). The protein synthesis levels with 4 hr treatments with WP and WP + CRE were greater compared to the CTL (<i>p</i> = 0.036 and <i>p</i> < 0.001, respectively), and 24 h levels were greater with WP versus other treatments (<i>p</i> < 0.05). p-p70S6K (Ser389) and p-rpS6 (Ser235/236) were greater with WP at 1 h compared to all other treatments (<i>p</i> < 0.05). No effects across time points were observed for p-mTOR (Ser2448), p-4E-BP1 (Thr37/46), or p-AMPKα (Thr172). <b>Conclusions</b>: WP increases protein synthesis and anabolic signaling with no additive effect from CRE. However, given that myotubes were not stressed nor stimulated to contract, such models are needed with the current treatment schematic to examine potential interactions. |
| format | Article |
| id | doaj-art-516fe8fd22f64a6ca6b64d4b1cde7d0d |
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| language | English |
| publishDate | 2025-05-01 |
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| spelling | doaj-art-516fe8fd22f64a6ca6b64d4b1cde7d0d2025-06-25T14:19:33ZengMDPI AGPhysiologia2673-94882025-05-01521710.3390/physiologia5020017The Effects of Creatine Monohydrate and/or Whey Protein on the Muscle Protein Synthesis and Anabolic Signaling Responses in Non-Stressed C<sub>2</sub>C<sub>12</sub> Murine MyotubesNicholas J. Kontos0Joshua S. Godwin1Anthony Agyin-Birikorang2Darren G. Candow3Christopher M. Lockwood4Michael D. Roberts5Christopher B. Mobley6Nutrabolt Applied and Molecular Physiology Laboratory, School of Kinesiology, Auburn University, Auburn, AL 36849, USANutrabolt Applied and Molecular Physiology Laboratory, School of Kinesiology, Auburn University, Auburn, AL 36849, USANutrabolt Applied and Molecular Physiology Laboratory, School of Kinesiology, Auburn University, Auburn, AL 36849, USAFaculty of Kinesiology and Health Studies, University of Regina, Regina, SK S7N 5A2, CanadaNutrabolt, Austin, TX 78749, USANutrabolt Applied and Molecular Physiology Laboratory, School of Kinesiology, Auburn University, Auburn, AL 36849, USANutrabolt Applied and Molecular Physiology Laboratory, School of Kinesiology, Auburn University, Auburn, AL 36849, USA<b>Background/Objectives</b>: Creatine monohydrate (CRE) is a popular nutritional supplement that increases lean/muscle mass accretion. Although data regarding CRE and its effects on muscle protein synthesis are mixed, we hypothesized that CRE may potentiate/extend the anabolic response to essential amino acids given that CRE acts as a high-energy phosphate buffer to potentially amplify anabolic signaling. Therefore, we used an in vitro model to determine whether CRE synergistically enhances myotube protein synthesis and the anabolic signaling responses to EAA-rich whey protein (WP). <b>Methods</b>: C<sub>2</sub>C<sub>12</sub> murine myotubes were treated with control media containing PBS (CTL), WP serum (5 mg/mL), CRE (10 mM), or WP + CRE. Myotubes were collected following 1, 4, and 24 h treatments (n = 6 replicates per treatment and time point) and assayed for relative creatine levels, myotube protein synthesis levels, and phosphorylation markers. <b>Results</b>: Cellular creatine levels were greater in CRE and WP + CRE versus CTL and WP at all treatment time points (<i>p</i> < 0.05). The protein synthesis levels with 4 hr treatments with WP and WP + CRE were greater compared to the CTL (<i>p</i> = 0.036 and <i>p</i> < 0.001, respectively), and 24 h levels were greater with WP versus other treatments (<i>p</i> < 0.05). p-p70S6K (Ser389) and p-rpS6 (Ser235/236) were greater with WP at 1 h compared to all other treatments (<i>p</i> < 0.05). No effects across time points were observed for p-mTOR (Ser2448), p-4E-BP1 (Thr37/46), or p-AMPKα (Thr172). <b>Conclusions</b>: WP increases protein synthesis and anabolic signaling with no additive effect from CRE. However, given that myotubes were not stressed nor stimulated to contract, such models are needed with the current treatment schematic to examine potential interactions.https://www.mdpi.com/2673-9488/5/2/17muscle protein synthesisanabolic signalingwhey proteincreatine |
| spellingShingle | Nicholas J. Kontos Joshua S. Godwin Anthony Agyin-Birikorang Darren G. Candow Christopher M. Lockwood Michael D. Roberts Christopher B. Mobley The Effects of Creatine Monohydrate and/or Whey Protein on the Muscle Protein Synthesis and Anabolic Signaling Responses in Non-Stressed C<sub>2</sub>C<sub>12</sub> Murine Myotubes Physiologia muscle protein synthesis anabolic signaling whey protein creatine |
| title | The Effects of Creatine Monohydrate and/or Whey Protein on the Muscle Protein Synthesis and Anabolic Signaling Responses in Non-Stressed C<sub>2</sub>C<sub>12</sub> Murine Myotubes |
| title_full | The Effects of Creatine Monohydrate and/or Whey Protein on the Muscle Protein Synthesis and Anabolic Signaling Responses in Non-Stressed C<sub>2</sub>C<sub>12</sub> Murine Myotubes |
| title_fullStr | The Effects of Creatine Monohydrate and/or Whey Protein on the Muscle Protein Synthesis and Anabolic Signaling Responses in Non-Stressed C<sub>2</sub>C<sub>12</sub> Murine Myotubes |
| title_full_unstemmed | The Effects of Creatine Monohydrate and/or Whey Protein on the Muscle Protein Synthesis and Anabolic Signaling Responses in Non-Stressed C<sub>2</sub>C<sub>12</sub> Murine Myotubes |
| title_short | The Effects of Creatine Monohydrate and/or Whey Protein on the Muscle Protein Synthesis and Anabolic Signaling Responses in Non-Stressed C<sub>2</sub>C<sub>12</sub> Murine Myotubes |
| title_sort | effects of creatine monohydrate and or whey protein on the muscle protein synthesis and anabolic signaling responses in non stressed c sub 2 sub c sub 12 sub murine myotubes |
| topic | muscle protein synthesis anabolic signaling whey protein creatine |
| url | https://www.mdpi.com/2673-9488/5/2/17 |
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