{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T05:46:12Z","timestamp":1782452772672,"version":"3.54.5"},"reference-count":49,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T00:00:00Z","timestamp":1781827200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key Research and Development Program of China","award":["2022YFF1100405"],"award-info":[{"award-number":["2022YFF1100405"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Background: Sarcopenia is an age-related disorder characterized by loss of muscle mass, strength, and function, driven by oxidative stress, chronic inflammation, and protein imbalance. Broccoli-derived peptides (BDP) exert anti-inflammatory and myofiber-protective effects, while leucine regulates energy metabolism and redox balance. Methods: We established a D-galactose aging mouse model and treated mice with BDP alone, leucine alone, or their combination for 8 weeks. Lean mass, muscle index, grip strength, endurance, and treadmill capacity were detected, and atrophic, disorganized myofibers were observed through histology. RNA-seq was applied to screen differential signaling pathways, and qPCR was used to verify related gene expression levels. Results: D-galactose caused marked deficits in lean mass, muscle index, grip strength, endurance, and treadmill capacity, accompanied by atrophic and disorganized myofibers. Single BDP or leucine partially reversed these deficits, but the combination produced the most robust improvements. RNA-seq revealed that BDP enriched actin, chemokine, and TNF pathways; leucine enriched Apelin and ECM pathways; while the combination uniquely regulated MAPK signaling. qPCR confirmed that co-administration optimally upregulated myogenic drivers (Myod1, Myog, Mef2c), suppressed catabolic\/inflammatory mediators (Mstn, Tnf, Cxcl10), and restored metabolic\/adhesive regulators (Sirt3, Aplnr, Icam1). Conclusions: BDP and leucine show superior efficacy in ameliorating sarcopenia, through multimodal regulation of multiple signaling pathways, offering a promising plant-based nutritional strategy against age-related muscle decline.<\/jats:p>","DOI":"10.3390\/nu18121997","type":"journal-article","created":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T13:04:04Z","timestamp":1781874244000},"page":"1997","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Broccoli-Derived Peptides and Leucine in Combination Ameliorate D-Galactose-Induced Sarcopenia in Mice"],"prefix":"10.3390","volume":"18","author":[{"given":"Kexin","family":"Yuan","sequence":"first","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-2290-4333","authenticated-orcid":false,"given":"Wenbin","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ning","family":"Su","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingyang","family":"Cui","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingyi","family":"Qi","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengyang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0421-0035","authenticated-orcid":false,"given":"Peng","family":"An","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8987-0533","authenticated-orcid":false,"given":"Junjie","family":"Luo","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7834-8742","authenticated-orcid":false,"given":"Yongting","family":"Luo","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Health, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.afos.2025.06.003","article-title":"Epidemiology of Sarcopenia: A Narrative Review","volume":"11","author":"Tseng","year":"2025","journal-title":"Osteoporos. Sarcopenia"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"659","DOI":"10.2147\/CIA.S517833","article-title":"Exploration of Pathogenesis and Cutting-Edge Treatment Strategies of Sarcopenia: A Narrative Review","volume":"20","author":"Gao","year":"2025","journal-title":"Clin. Interv. Aging"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"389","DOI":"10.5720\/kjcn.2025.00360","article-title":"Dietary Factors and Nutritional Guidelines for Sarcopenia in Older Adults: A Narrative Review","volume":"30","author":"Heo","year":"2025","journal-title":"Korean J. Community Nutr."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1002\/jcsm.12651","article-title":"Sarcopenia: Prevalence, Associated Factors, and the Risk of Mortality and Disability in Japanese Older Adults","volume":"12","author":"Kitamura","year":"2021","journal-title":"J. Cachexia Sarcopenia Muscle"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"20230165","DOI":"10.1002\/EXP.20230165","article-title":"Exploring Precision Treatments in Immune-Mediated Inflammatory Diseases: Harnessing the Infinite Potential of Nucleic Acid Delivery","volume":"5","author":"Xu","year":"2025","journal-title":"Exploration"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Damiano, S., Muscariello, E., La Rosa, G., Di Maro, M., Mondola, P., and Santillo, M. (2019). Dual Role of Reactive Oxygen Species in Muscle Function: Can Antioxidant Dietary Supplements Counteract Age-Related Sarcopenia?. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20153815"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"757","DOI":"10.2147\/CIA.S158513","article-title":"Oxidative Stress, Aging, and Diseases","volume":"13","author":"Liguori","year":"2018","journal-title":"Clin. Interv. Aging"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"20210267","DOI":"10.1002\/EXP.20210267","article-title":"Bioinspired Copper Single-Atom Nanozyme as a Superoxide Dismutase-Like Antioxidant for Sepsis Treatment","volume":"4","author":"Yang","year":"2022","journal-title":"Exploration"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Hajam, Y.A., Rani, R., Ganie, S.Y., Sheikh, T.A., Javaid, D., Qadri, S.S., Pramodh, S., Alsulimani, A., Alkhanani, M.F., and Harakeh, S. (2022). Oxidative Stress in Human Pathology and Aging: Molecular Mechanisms and Perspectives. Cells, 11.","DOI":"10.3390\/cells11030552"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Wang, H., Zhang, Y., Wang, X., Qiao, Z., Wang, J., Li, Y., and Sun, Y. (2025). \u03b1-Ketoglutarate Ameliorates Sarcopenia in D-Galactose-Induced Aging Mice by Modulating Protein Homeostasis and Optimizing Mitochondrial Function. Nutrients, 17.","DOI":"10.3390\/nu17213336"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"102543","DOI":"10.1016\/j.arr.2024.102543","article-title":"Rodent Model Intervention for Prevention and Optimal Management of Sarcopenia: A Systematic Review on the Beneficial Effects of Nutrients & Non-Nutrients and Exercise to Improve Skeletal Muscle Health","volume":"102","author":"Mayakrishnan","year":"2024","journal-title":"Ageing Res. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Adebayo, S.D., Poddar, S., and Yu, J. (2026). Antihypertensive Peptides and Hydrolysates Derived from Plant Proteins and Their Bioavailability. Foods, 15.","DOI":"10.3390\/foods15050900"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"144440","DOI":"10.1016\/j.foodchem.2025.144440","article-title":"Enhancing Bioavailability and Functionality of Plant Peptides and Proteins: A Review of Novel Strategies for Food and Pharmaceutical Applications","volume":"485","author":"Shadrack","year":"2025","journal-title":"Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1849","DOI":"10.1016\/j.clnu.2023.08.010","article-title":"A Focus on Leucine in the Nutritional Regulation of Human Skeletal Muscle Metabolism in Ageing, Exercise and Unloading States","volume":"42","author":"Ely","year":"2023","journal-title":"Clin. Nutr."},{"key":"ref_15","first-page":"2616s","article-title":"Leucine and Mammalian Target of Rapamycin-Dependent Activation of Muscle Protein Synthesis in Aging","volume":"146","year":"2026","journal-title":"J. Nutr."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"796","DOI":"10.1016\/j.tem.2016.06.010","article-title":"Amino Acid Sensing in Skeletal Muscle","volume":"27","author":"Moro","year":"2016","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1089\/jmf.2021.0094","article-title":"Mtor1c Activation with the Leucine \u201cTrigger\u201d for Prevention of Sarcopenia in Older Adults during Lockdown","volume":"25","author":"Mai","year":"2022","journal-title":"J. Med. Food"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3811","DOI":"10.1007\/s11255-025-04560-9","article-title":"Anti-Sarcopenic Effect of Leucine-Enriched Branched-Chain Amino Acid Supplementation among Elderly Chronic Kidney Disease Patients: A Double-Blinded Randomized Controlled Trial","volume":"57","author":"Sunsandee","year":"2025","journal-title":"Int. Urol. Nephrol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Huang, C., and Hsieh, M.H. (2025). Effects of Leucine Supplementation in Older Adults with Sarcopenia: A Meta-Analysis. Nutrients, 17.","DOI":"10.3390\/nu17152413"},{"key":"ref_20","first-page":"75","article-title":"Dietary Leucine Intake and Sarcopenia: From Isolated Supplementation to Combined Strategies","volume":"29","author":"Walrand","year":"2026","journal-title":"Curr. Opin. Clin. Nutr. Metab. Care"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5419","DOI":"10.1021\/acs.jafc.2c09094","article-title":"Potential of Food Protein-Derived Bioactive Peptides against Sarcopenia: A Comprehensive Review","volume":"71","author":"Zhu","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"20210089","DOI":"10.1002\/EXP.20210089","article-title":"Advanced Bioactive Nanomaterials for Biomedical Applications","volume":"1","author":"Zhao","year":"2021","journal-title":"Exploration"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Szerszunowicz, I., and Kozicki, S. (2023). Plant-Derived Proteins and Peptides as Potential Immunomodulators. Molecules, 29.","DOI":"10.3390\/molecules29010209"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.1002\/ptr.8420","article-title":"Plant-Derived Treatments for Different Types of Muscle Atrophy","volume":"39","author":"Wang","year":"2025","journal-title":"Phytother. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"104283","DOI":"10.1016\/j.drudis.2024.104283","article-title":"Repairing Muscle with Broccoli-Derived Sulforaphane: A Preclinical Evaluation for the Treatment of Mitochondrial Myopathies","volume":"30","author":"Lilley","year":"2025","journal-title":"Drug Discov. Today"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Xie, C., Yan, R., and Tao, R. (2026). Combined Resistance Training and Amino Acid-Based Supplementation for Sarcopenia in Older Adults: A Systematic Review and Meta-Analysis. BMC Musculoskelet. Disord., 27.","DOI":"10.1186\/s12891-025-09436-8"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Fang, J., Yan, W., Sun, X., and Chen, J. (2025). The Role of Exercise-Induced Short-Chain Fatty Acids in the Gut-Muscle Axis: Implications for Sarcopenia Prevention and Therapy. Front. Microbiol., 16.","DOI":"10.3389\/fmicb.2025.1665551"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Kwon, R.J., Mijan, M.A., Son, S.M., Yoo, W., and Kim, T. (2025). Combined Protein, Probiotics, and Exercise Therapy for Sarcopenia: A Comprehensive Review. Cells, 14.","DOI":"10.3390\/cells14171375"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3278","DOI":"10.1007\/s12011-023-03898-4","article-title":"The Effectiveness of Time in Treatment with Vitamin C and Broccoli Extract on Cadmium Poisoning in Mice: Histological Changes of Testicular Tissue and Cell Apoptotic Index","volume":"202","author":"Raeeszadeh","year":"2024","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Liao, Y., Li, D., Zhou, X., Peng, Z., Meng, Z., Liu, R., and Yang, W. (2021). Pyruvate Might Bridge Gut Microbiota and Muscle Health in Aging Mice after Chronic High Dose of Leucine Supplementation. Front. Med., 8.","DOI":"10.3389\/fmed.2021.755803"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Wu, W., Guo, X., Qu, T., Huang, Y., Tao, J., He, J., Wang, X., Luo, J., An, P., and Zhu, Y. (2024). The Combination of Lactoferrin and Creatine Ameliorates Muscle Decay in a Sarcopenia Murine Model. Nutrients, 16.","DOI":"10.3390\/nu16121958"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"7339","DOI":"10.18632\/aging.101733","article-title":"Leonurine Ameliorates D-Galactose-Induced Aging in Mice through Activation of the Nrf2 Signalling Pathway","volume":"11","author":"Chen","year":"2019","journal-title":"Aging"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"20230098","DOI":"10.1002\/EXP.20230098","article-title":"Decoding the Regulatory Role of Atp Synthase Inhibitory Factor 1 (Atpif1) in Wallerian Degeneration and Peripheral Nerve Regeneration","volume":"4","author":"Qian","year":"2024","journal-title":"Exploration"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1182","DOI":"10.3168\/jds.2024-25532","article-title":"Bioactive Milk-Derived Nutrient Mfg-E8 Ameliorates Skeletal Muscle Atrophy Induced by Mitochondria Damage in Aging Rats Via Activating the Mapk\/Erk Signaling Pathway","volume":"108","author":"Guan","year":"2025","journal-title":"J. Dairy Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1097","DOI":"10.1007\/s40279-024-02007-2","article-title":"Exercise-Regulated Mitochondrial and Nuclear Signalling Networks in Skeletal Muscle","volume":"54","author":"Reisman","year":"2024","journal-title":"Sports Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1101\/gad.1281105","article-title":"An Initial Blueprint for Myogenic Differentiation","volume":"15","author":"Blais","year":"2005","journal-title":"Genes Dev."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"111931","DOI":"10.1016\/j.exger.2022.111931","article-title":"Inflammaging: The Ground for Sarcopenia?","volume":"168","author":"Liang","year":"2022","journal-title":"Exp. Gerontol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"114513","DOI":"10.1016\/j.ygcen.2024.114513","article-title":"The Role of Tgf-\u0392 Signaling in Muscle Atrophy, Sarcopenia and Cancer Cachexia","volume":"353","author":"Lan","year":"2024","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1007\/s11357-022-00715-z","article-title":"Altered T Cell Infiltration and Enrichment of Leukocyte Regulating Pathways within Aged Skeletal Muscle Are Associated Impaired Muscle Function Following Influenza Infection","volume":"45","author":"Keilich","year":"2023","journal-title":"Geroscience"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5135","DOI":"10.7150\/ijbs.115518","article-title":"Roles of Sirt3 in Aging and Aging-Related Diseases","volume":"21","author":"You","year":"2025","journal-title":"Int. J. Biol. Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1360","DOI":"10.1038\/s41591-018-0131-6","article-title":"The Exerkine Apelin Reverses Age-Associated Sarcopenia","volume":"24","author":"Vinel","year":"2018","journal-title":"Nat. Med."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Buckley, K.H., Nestor-Kalinoski, A.L., and Pizza, F.X. (2022). Intercellular Adhesion Molecule-1 Enhances Myonuclear Transcription during Injury-Induced Muscle Regeneration. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23137028"},{"key":"ref_43","first-page":"101955","article-title":"Effects of alpha-ketoisocaproate in oxidative stress-induced C2C12 myotubes via inhibition of p38 MAPK and ERK1\/2","volume":"41","author":"Kim","year":"2025","journal-title":"Biochem. Biophys. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1089\/jmf.2025.k.0050","article-title":"Phlomis umbrosa Turcz. water extract attenuates dexamethasone-induced muscle atrophy by regulating PI3K\/Akt\/mTOR pathway","volume":"28","author":"Lee","year":"2025","journal-title":"J. Med. Food"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"de Souza, A.L.G., Alves, A.L.R., Martinez, C.G., de Sousa, J.C., and Kurtenbach, E. (2025). Biomarkers of skeletal muscle atrophy based on Atrogenes evaluation: A systematic review and meta-analysis study. Int. J. Mol. Sci., 26.","DOI":"10.37766\/inplasy2025.2.0089"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1186\/s12979-024-00458-9","article-title":"Harnessing Immunomodulation to Combat Sarcopenia: Current Insights and Possible Approaches","volume":"21","author":"Zhang","year":"2024","journal-title":"Immun. Ageing"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"123205","DOI":"10.1016\/j.lfs.2024.123205","article-title":"Activation of the Apelin\/Apj System by Vitamin D Attenuates Age-Related Muscle Atrophy","volume":"359","author":"Lee","year":"2024","journal-title":"Life Sci."},{"key":"ref_48","first-page":"404","article-title":"Extracellular Matrix in Skeletal Muscle Injury and Atrophy: Mechanisms and Therapeutic Implications","volume":"52","author":"Ge","year":"2025","journal-title":"J. Orthop. Transl."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"3219","DOI":"10.7150\/ijbs.96885","article-title":"Unlocking the Nexus of Sirtuins: A Comprehensive Review of Their Role in Skeletal Muscle Metabolism, Development, and Disorders","volume":"20","author":"Gibril","year":"2024","journal-title":"Int. J. Biol. Sci."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/18\/12\/1997\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T04:55:30Z","timestamp":1782449730000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/18\/12\/1997"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,19]]},"references-count":49,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["nu18121997"],"URL":"https:\/\/doi.org\/10.3390\/nu18121997","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,19]]}}}