{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T06:57:29Z","timestamp":1780556249638,"version":"3.54.1"},"reference-count":45,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2023,11,5]],"date-time":"2023-11-05T00:00:00Z","timestamp":1699142400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"JSPS KAKENHI","doi-asserted-by":"publisher","award":["23K05095"],"award-info":[{"award-number":["23K05095"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Glucocorticoids, commonly used to manage inflammatory diseases, can induce muscle atrophy by accelerating the breakdown of muscle proteins. This research delves into the influence of Prolyl-hydroxyproline (Pro-Hyp), a collagen-derived peptide, on muscle atrophy induced with dexamethasone (DEX), a synthetic glucocorticoid, in mouse C2C12 skeletal myotubes. Exposure to DEX (10 \u03bcM) for 6 days resulted in a decrease in myotube diameter, along with elevated mRNA and protein levels of two muscle-atrophy-related ubiquitin ligases, muscle atrophy F-box (MAFbx, also known as atrogin-1) and muscle ring finger 1 (MuRF-1). Remarkably, treatment with 0.1 mM of Pro-Hyp mitigated the reduction in myotube thickness caused by DEX, while promoting the phosphorylation of Akt, mammalian target of rapamycin (mTOR), and forkhead box O3a (Foxo3a). This led to the inhibition of the upregulation of the ubiquitin ligases atrogin-1 and MuRF-1. These findings indicate the potential significance of Pro-Hyp as a promising therapeutic target for countering DEX-induced muscle atrophy.<\/jats:p>","DOI":"10.3390\/biom13111617","type":"journal-article","created":{"date-parts":[[2023,11,5]],"date-time":"2023-11-05T07:35:06Z","timestamp":1699169706000},"page":"1617","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Preventive Effects of Collagen-Derived Dipeptide Prolyl-Hydroxyproline against Dexamethasone-Induced Muscle Atrophy in Mouse C2C12 Skeletal Myotubes"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5134-2519","authenticated-orcid":false,"given":"Yoshifumi","family":"Kimira","sequence":"first","affiliation":[{"name":"Department of Clinical Dietetics and Human Nutrition, Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado-shi 350-0295, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Konosuke","family":"Osawa","sequence":"additional","affiliation":[{"name":"Department of Clinical Dietetics and Human Nutrition, Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado-shi 350-0295, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshihiro","family":"Osawa","sequence":"additional","affiliation":[{"name":"Department of Clinical Dietetics and Human Nutrition, Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado-shi 350-0295, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroshi","family":"Mano","sequence":"additional","affiliation":[{"name":"Department of Clinical Dietetics and Human Nutrition, Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado-shi 350-0295, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1007\/s13539-012-0074-6","article-title":"Molecular and Cellular Mechanisms of Skeletal Muscle Atrophy: An Update","volume":"3","author":"Fanzani","year":"2012","journal-title":"J. Cachexia Sarcopenia Muscle"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1364","DOI":"10.1161\/01.CIR.85.4.1364","article-title":"Contribution of Skeletal Muscle Atrophy to Exercise Intolerance and Altered Muscle Metabolism in Heart Failure","volume":"85","author":"Mancini","year":"1992","journal-title":"Circulation"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Dubashynskaya, N.V., Bokatyi, A.N., and Skorik, Y.A. (2021). Dexamethasone Conjugates: Synthetic Approaches and Medical Prospects. Biomedicines, 9.","DOI":"10.3390\/biomedicines9040341"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2163","DOI":"10.1016\/j.biocel.2013.05.036","article-title":"Glucocorticoid-Induced Skeletal Muscle Atrophy","volume":"45","author":"Schakman","year":"2013","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"E469","DOI":"10.1152\/ajpendo.00204.2014","article-title":"Skeletal Muscle Atrophy and the E3 Ubiquitin Ligases MuRF1 and MAFbx\/Atrogin-1","volume":"307","author":"Bodine","year":"2014","journal-title":"Am. J. Physiol. Endocrinol. Metab."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1016\/S1097-2765(04)00211-4","article-title":"The IGF-1\/PI3K\/Akt Pathway Prevents Expression of Muscle Atrophy-Induced Ubiquitin Ligases by Inhibiting FOXO Transcription Factors","volume":"14","author":"Stitt","year":"2004","journal-title":"Mol. Cell"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2093","DOI":"10.1080\/09168451.2016.1210502","article-title":"Cortisone and Dexamethasone Inhibit Myogenesis by Modulating the AKT\/MTOR Signaling Pathway in C2C12","volume":"80","author":"Kim","year":"2016","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Ricard-Blum, S. (2011). The Collagen Family. Cold Spring Harb. Perspect. Biol., 3.","DOI":"10.1101\/cshperspect.a004978"},{"key":"ref_9","first-page":"49","article-title":"The Collagen Superfamily\u2014Diverse Structures and Assemblies","volume":"27","author":"Hulmes","year":"1992","journal-title":"Essays Biochem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/S0968-4328(00)00042-1","article-title":"Collagen Structure and Functional Implications","volume":"32","author":"Ottani","year":"2001","journal-title":"Micron"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1515\/CCLM.2008.259","article-title":"Prolyl-Hydroxyproline Dipeptide in Non-Hydrolyzed Morning Urine and Its Value in Postmenopausal Osteoporosis","volume":"46","author":"Husek","year":"2008","journal-title":"Clin. Chem. Lab. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1356","DOI":"10.1080\/09168451.2015.1027653","article-title":"Detection of Endogenous and Food-Derived Collagen Dipeptide Prolylhydroxyproline (Pro-Hyp) in Allergic Contact Dermatitis-Affected Mouse Ear","volume":"79","author":"Kusubata","year":"2015","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1016\/S0945-053X(97)90030-5","article-title":"The Collagen Triple-Helix Structure","volume":"15","author":"Brodsky","year":"1997","journal-title":"Matrix Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"6531","DOI":"10.1021\/jf050206p","article-title":"Identification of Food-Derived Collagen Peptides in Human Blood after Oral Ingestion of Gelatin Hydrolysates","volume":"53","author":"Iwai","year":"2005","journal-title":"J. Agric. Food Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1021\/jf9031487","article-title":"Absorption and Effectiveness of Orally Administered Low Molecular Weight Collagen Hydrolysate in Rats","volume":"58","author":"Shimizu","year":"2010","journal-title":"J. Agric. Food Chem."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"K\u00f6nig, D., Oesser, S., Scharla, S., Zdzieblik, D., and Gollhofer, A. (2018). Specific Collagen Peptides Improve Bone Mineral Density and Bone Markers in Postmenopausal Women\u2014A Randomized Controlled Study. Nutrients, 10.","DOI":"10.3390\/nu10010097"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2221","DOI":"10.1185\/030079906X148373","article-title":"Collagen Hydrolysate for the Treatment of Osteoarthritis and Other Joint Disorders: A Review of the Literature","volume":"22","author":"Bello","year":"2006","journal-title":"Curr. Med. Res. Opin."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1139\/apnm-2016-0390","article-title":"Improvement of Activity-Related Knee Joint Discomfort Following Supplementation of Specific Collagen Peptides","volume":"42","author":"Zdzieblik","year":"2017","journal-title":"Appl. Physiol. Nutr. Metab."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1021\/jf802785h","article-title":"Effect of Prolyl-Hydroxyproline (Pro-Hyp), a Food-Derived Collagen Peptide in Human Blood, on Growth of Fibroblasts from Mouse Skin","volume":"57","author":"Shigemura","year":"2009","journal-title":"J. Agric. Food Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1016\/j.bbrc.2014.09.121","article-title":"Collagen-Derived Dipeptide Prolyl-Hydroxyproline Promotes Differentiation of MC3T3-E1 Osteoblastic Cells","volume":"453","author":"Kimira","year":"2014","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Ide, K., Takahashi, S., Sakai, K., Taga, Y., Ueno, T., Dickens, D., Jenkins, R., Falciani, F., Sasaki, T., and Ooi, K. (2021). The Dipeptide Prolyl-Hydroxyproline Promotes Cellular Homeostasis and Lamellipodia-Driven Motility via Active \u03921-Integrin in Adult Tendon Cells. J. Biol. Chem., 297.","DOI":"10.1016\/j.jbc.2021.100819"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1620","DOI":"10.1016\/j.joca.2009.07.001","article-title":"Chondroprotective Effect of the Bioactive Peptide Prolyl-Hydroxyproline in Mouse Articular Cartilage in Vitro and in Vivo","volume":"17","author":"Nakatani","year":"2009","journal-title":"Osteoarthr. Cartil."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Kimira, Y., Sato, T., Sakamoto, M., Osawa, Y., and Mano, H. (2023). Collagen-Derived Dipeptide Pro-Hyp Enhanced ATDC5 Chondrocyte Differentiation under Hypoxic Conditions. Molecules, 28.","DOI":"10.3390\/molecules28124664"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1292","DOI":"10.1016\/j.bbrc.2016.08.114","article-title":"The Collagen Derived Dipeptide Hydroxyprolyl-Glycine Promotes C2C12 Myoblast Differentiation and Myotube Hypertrophy","volume":"478","author":"Kitakaze","year":"2016","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Jendricke, P., Centner, C., Zdzieblik, D., Gollhofer, A., and K\u00f6nig, D. (2019). Specific Collagen Peptides in Combination with Resistance Training Improve Body Composition and Regional Muscle Strength in Premenopausal Women: A Randomized Controlled Trial. Nutrients, 11.","DOI":"10.3390\/nu11040892"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Zdzieblik, D., Jendricke, P., Oesser, S., Gollhofer, A., and K\u00f6nig, D. (2021). The Influence of Specific Bioactive Collagen Peptides on Body Composition and Muscle Strength in Middle-Aged, Untrained Men: A Randomized Controlled Trial. Int. J. Environ. Res. Public Health, 18.","DOI":"10.3390\/ijerph18094837"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1237","DOI":"10.1017\/S0007114515002810","article-title":"Collagen Peptide Supplementation in Combination with Resistance Training Improves Body Composition and Increases Muscle Strength in Elderly Sarcopenic Men: A Randomised Controlled Trial","volume":"114","author":"Zdzieblik","year":"2015","journal-title":"Br. J. Nutr."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Oh, J., Park, S.H., Kim, D.S., Choi, W., Jang, J., Rahmawati, L., Jang, W.Y., Lim, H.K., Hwang, J.Y., and Gu, G.R. (2023). The Preventive Effect of Specific Collagen Peptides against Dexamethasone-Induced Muscle Atrophy in Mice. Molecules, 28.","DOI":"10.3390\/molecules28041950"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Kim, J.E., Kwon, E.Y., and Han, Y. (2022). A Collagen Hydrolysate Containing Tripeptides Ameliorates Sarcopenia in Middle-Aged Mice. Molecules, 27.","DOI":"10.3390\/molecules27092718"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Kimira, Y., Odaira, H., Nomura, K., Taniuchi, Y., Inoue, N., Nakatani, S., Shimizu, J., Wada, M., and Mano, H. (2017). Collagen-Derived Dipeptide Prolyl-Hydroxyproline Promotes Osteogenic Differentiation through Foxg1. Cell. Mol. Biol. Lett., 22.","DOI":"10.1186\/s11658-017-0060-2"},{"key":"ref_31","first-page":"479","article-title":"Matrine Improves Skeletal Muscle Atrophy by Inhibiting E3 Ubiquitin Ligases and Activating the Akt\/MTOR\/FoxO3\u03b1 Signaling Pathway in C2C12 Myotubes and Mice","volume":"42","author":"Chen","year":"2019","journal-title":"Oncol. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1007\/s00424-020-02367-4","article-title":"Irisin Prevents Dexamethasone-Induced Atrophy in C2C12 Myotubes","volume":"472","author":"Chang","year":"2020","journal-title":"Pflug. Arch."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"758","DOI":"10.1126\/science.2414846","article-title":"Plasticity of the Differentiated State","volume":"230","author":"Blau","year":"1985","journal-title":"Science"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"14440","DOI":"10.1073\/pnas.251541198","article-title":"Atrogin-1, a Muscle-Specific F-Box Protein Highly Expressed during Muscle Atrophy","volume":"98","author":"Gomes","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1266","DOI":"10.1038\/emboj.2008.52","article-title":"The Initiation Factor EIF3-f Is a Major Target for Atrogin1\/MAFbx Function in Skeletal Muscle Atrophy","volume":"27","author":"Offner","year":"2008","journal-title":"EMBO J."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1016\/j.cmet.2007.09.009","article-title":"The E3 Ligase MuRF1 Degrades Myosin Heavy Chain Protein in Dexamethasone-Treated Skeletal Muscle","volume":"6","author":"Clarke","year":"2007","journal-title":"Cell Metab."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1495","DOI":"10.1016\/j.metabol.2013.05.018","article-title":"Suppression of Atrogin-1 and MuRF1 Prevents Dexamethasone-Induced Atrophy of Cultured Myotubes","volume":"62","author":"Castillero","year":"2013","journal-title":"Metabolism"},{"key":"ref_38","first-page":"235","article-title":"Involvement of Myogenic Regulator Factors during Fusion in the Cell Line C2C12","volume":"46","author":"Dedieu","year":"2002","journal-title":"Int. J. Dev. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1302","DOI":"10.1002\/jcb.22360","article-title":"Expression and Subcellular Localization of Myogenic Regulatory Factors during the Differentiation of Skeletal Muscle C2C12 Myoblasts","volume":"108","author":"Ferri","year":"2009","journal-title":"J. Cell. Biochem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"32081","DOI":"10.1074\/jbc.M502876200","article-title":"Akt Activates the Mammalian Target of Rapamycin by Regulating Cellular ATP Level and AMPK Activity","volume":"280","author":"Nogueira","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"6670","DOI":"10.1038\/ncomms7670","article-title":"Regulation of Autophagy and the Ubiquitin\u2013Proteasome System by the FoxO Transcriptional Network during Muscle Atrophy","volume":"6","author":"Milan","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1016\/S0092-8674(04)00400-3","article-title":"Foxo Transcription Factors Induce the Atrophy-Related Ubiquitin Ligase Atrogin-1 and Cause Skeletal Muscle Atrophy","volume":"117","author":"Sandri","year":"2004","journal-title":"Cell"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Nomura, K., Kimira, Y., Osawa, Y., Kataoka-Matsushita, A., Takao, K., Sugita, Y., Shimizu, J., Wada, M., and Mano, H. (2021). Stimulation of the Runx2 P1 Promoter by Collagen-Derived Dipeptide Prolyl-Hydroxyproline Bound to Foxg1 and Foxo1 in Osteoblasts. Biosci. Rep., 41.","DOI":"10.1042\/BSR20210304"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2121","DOI":"10.1016\/j.biocel.2013.04.023","article-title":"Protein Breakdown in Muscle Wasting: Role of Autophagy-Lysosome and Ubiquitin-Proteasome","volume":"45","author":"Sandri","year":"2013","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"11126","DOI":"10.3390\/ijms150611126","article-title":"TRAF6 Inhibition Rescues Dexamethasone-Induced Muscle Atrophy","volume":"15","author":"Sun","year":"2014","journal-title":"Int. J. Mol. Sci."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/11\/1617\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:17:46Z","timestamp":1760131066000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/11\/1617"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,5]]},"references-count":45,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2023,11]]}},"alternative-id":["biom13111617"],"URL":"https:\/\/doi.org\/10.3390\/biom13111617","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,11,5]]}}}