Sains Malaysiana 55(10)(2026): 1664-1680

http://doi.org/10.17576/jsm-2026-5510-06

 

Untargeted Muscle Tissue Metabolites Profiling in an Ageing Animal Model Supplemented with Ginger Active Compounds

(Profil Metabolit Tisu Otot Tidak Bersasar dalam Model Haiwan Penuaan yang Disuplementasi dengan Sebatian Aktif Halia)

 

NUR FATIN NABILAH MOHD SAHARDI¹, NUR FATHIAH ABDUL SANI², JEN KIT TAN^2,4, MARIAM FIRDHAUS MAD NORDIN³ & SUZANA MAKPOL^2,4,*

 

¹Secretariat Research and Innovation, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur, Malaysia

²Department of Biochemistry, Faculty of Medicine, Level 17 Preclinical Building, Universiti KebangsaanMalaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur, Malaysia

³AM Zaideen Ventures Sdn Bhd, Kuala Lumpur, Malaysia

⁴Ageing and Degenerative Diseases Research Group, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

 

Received: 22 April 2026/Accepted: 23 September 2026

 

*Corresponding author; email: suzanamakpol@ukm.edu.my

 

Abstract

Age-related muscle loss is a significant health concern among older adults, contributing to frailty, reduced strength, and functional decline, with oxidative stress identified as a key underlying driver of this deterioration. Ginger (Zingiber officinale Roscoe), a traditional herb with established antioxidant properties, has shown potential to mitigate oxidative damage. However, its effects on muscle metabolic regulation during the ageing process remain largely unexplored. Therefore, this study aimed to investigate the effects of ginger extract on metabolic pathways associated with age-related muscle loss. Sprague-Dawley rats of various ages (young, adult, and old) were administered distilled water or ginger extract (200 mg/kg body weight) daily for three months. Muscle tissues were subjected to untargeted metabolomic profiling using UHPLC-MS/MS. Correlation analyses were conducted to determine associations between significantly altered metabolites and oxidative stress and inflammatory markers. Untargeted metabolomic profiling showed that ageing markedly altered muscle metabolites, particularly those in the phenylalanine, tyrosine, and tryptophan biosynthesis pathway. Ginger supplementation partially reversed these age-associated shifts; most notably, it significantly downregulated β-muricholic acid and xanthosine in old rats, which had increased during ageing. Furthermore, ginger treatment upregulated γ-L-glutamyl-L-glutamic acid, suggesting enhanced antioxidant capacity and muscle regenerative pathways. These findings suggest that ginger extract may attenuate age-related physiological decline by modulating several key metabolites disrupted during ageing, highlighting its potential as a supplement to support healthy ageing in older individuals.

Keywords: Ginger extract; metabolomic study; muscle ageing; oxidative stress

 

Abstrak

Kehilangan otot berkaitan usia merupakan isu kesihatan yang signifikan dalam kalangan warga emas yang menyumbang kepada kerapuhan, penurunan kekuatan dan kemerosotan fungsi, dengan tekanan oksidatif dikenal pasti sebagai pemacu utama kepada kemerosotan otot. Halia (Zingiber officinale Roscoe), sejenis herba tradisional yang mempunyai sifat antioksidan yang terbukti, telah menunjukkan potensi dalam mengurangkan kerosakan oksidatif. Walau bagaimanapun, kesannya terhadap regulasi metabolik otot semasa proses penuaan masih belum banyak dikaji. Oleh itu, penyelidikan ini bertujuan untuk mengkaji kesan ekstrak halia terhadap laluan metabolik yang berkaitan dengan kehilangan otot akibat penuaan. Tikus Sprague-Dawley daripada pelbagai peringkat umur (muda, dewasa dan tua) diberikan air suling atau ekstrak halia (200 mg/kg berat badan) setiap hari selama tiga bulan. Tisu otot dianalisis menggunakan pemprofilan metabolik tidak bersasar dengan UHPLC-MS/MS. Analisis korelasi dijalankan untuk menentukan hubungan antara metabolit yang berubah secara signifikan dengan penanda tekanan oksidatif dan keradangan. Hasil pemprofilan metabolomik tidak bersasar menunjukkan bahawa penuaan secara ketara mengubah metabolit otot, khususnya dalam laluan biosintesis fenilalanina, tirosin dan triptofan. Suplementasi halia didapati sebahagiannya membalikkan perubahan berkaitan usia ini; yang paling ketara, ia menurunkan secara signifikan asid β-murikolik dan xantosin dalam tikus tua, yang meningkat semasa proses penuaan. Selain itu, rawatan halia meningkatkan γ-L-glutamil-L-asid-glutamik, yang mencadangkan peningkatan kapasiti antioksidan dan laluan regenerasi otot. Keputusan ini menunjukkan bahawa ekstrak halia berpotensi memperlahankan kemerosotan fisiologi berkaitan usia dengan memodulasi beberapa metabolit utama yang terganggu semasa proses penuaan, sekali gus menonjolkan potensinya sebagai suplemen untuk menyokong penuaan sihat dalam kalangan individu berusia.

Kata kunci: Ekstrak halia; kajian metabolik; penuaan otot; tekanan oksidatif

 

REFERENCES

Abdul Sani, N.F., Belani, L.K., Pui Sin, C., Abdul Rahman, S.N.A., Das, S., Zar Chi, T., Makpol, S. & Yusof, Y.A.M. 2014. Effect of the combination of gelam honey and ginger on oxidative stress and metabolic profile in streptozotocin-induced diabetic Sprague-Dawley rats. BioMed Research International 2014: 160695.

Aggarwal, R. & Bains, K. 2022. Protein, lysine and vitamin D: Critical role in muscle and bone health. Critical Reviews in Food Science and Nutrition 62(9): 2548-2559.

Ansoleaga, B., Jové, M., Schlüter, A., Garcia-Esparcia, P., Moreno, J., Pujol, A., Pamplona, R., Portero-Otín, M. & Ferrer, I. 2015. Deregulation of purine metabolism in Alzheimer's disease. Neurobiology of Aging 36(1): 68-80.

Cannataro, R., Carbone, L., Petro, J.L., Cione, E., Vargas, S., Angulo, H., Forero, D.A., Odriozola-Martínez, A., Kreider, R.B. & Bonilla, D.A. 2021. Sarcopenia: Etiology, nutritional approaches, and miRNAs. International Journal of Molecular Sciences 22(18): 9724.

Chae, M., Park, H.S. & Park, K. 2021. Association between dietary branched-chain amino acid intake and skeletal muscle mass index among Korean adults: Interaction with obesity. Nutrition Research and Practice 15(2): 203-212.

Cleminson, J.R., Stuart, A.L., Pasco, J.A., Hodge, J.M., Berk, M., Samarasinghe, R.M. & Williams, L.J. 2020. Dietary tryptophan and bone health: A cross-sectional, population-based study. Archives of Osteoporosis 15: 167.

Cruz-Jentoft, A.J. & Sayer, A.A. 2019. Sarcopenia. Lancet 393(10191): 2636-2646.

Cruz-Jentoft, A.J., Bahat, G., Bauer, J., Boirie, Y., Bruyère, O., Cederholm, T., Cooper, C., Landi, F., Rolland, Y., Sayer, A.A., Schneider, S.M., Sieber, C.C., Topinkova, E., Vandewoude, M., Visser, M. & Zamboni, M. 2019. Sarcopenia: Revised European consensus on definition and diagnosis. Age and Ageing 48(4): 16-31.

Danial Efendy Goon, Sharaniza Ab-Rahim, Amir Hakimi Mohd Sakri, Musalmah Mazlan, Tan, Jen Kit, Mardiana Abdul Aziz, Mohd Noor Norizal, Effendi Ibrahim & Siti Hamimah Sheikh Abdul Kadir. 2021. Untargeted serum metabolites profiling in high-fat diet mice supplemented with enhanced palm tocotrienol-rich fraction using UHPLC-MS. Scientific Reports 11(1): 21001.

Dao, T., Green, A.E., Kim, Y.A., Bae, S.J., Ha, K.T., Gariani, K., Lee, M.R., Menzies, K.J. & Ryu, D. 2020. Sarcopenia and muscle aging: A brief overview. Endocrinology and Metabolism 35(4): 716-732.

Drey, M. 2011. Sarcopenia - Pathophysiology and clinical relevance. Wiener Medizinische Wochenschrift 161(17): 402-408.

Duan, Y., Zeng, L., Li, F., Wang, W., Li, Y., Guo, Q., Ji, Y., Tan, B.E. & Yin, Y. 2017. Effect of branched-chain amino acid ratio on the proliferation, differentiation, and expression levels of key regulators involved in protein metabolism of myocytes. Nutrition 36(1): 8-16.

Dunn, W.B., Broadhurst, D., Begley, P., Zelena, E., Francis-Mcintyre, S., Anderson, N., Brown, M., Knowles, J.D., Halsall, A., Haselden, J.N., Nicholls, A.W., Wilson, I.D., Kell, D.B. & Goodacre, R. 2011. Procedures for large-scale metabolic profiling of serum and plasma using gas chromatography and liquid chromatography coupled to mass spectrometry. Nature Protocols 6(7): 1060-1083.

Elhassan, Y.S., Kluckova, K., Fletcher, R.S., Schmidt, M.S., Garten, A., Doig, C.L., Cartwright, D.M., Oakey, L., Burley, C.V., Jenkinson, N., Wilson, M., Lucas, S.J.E., Akerman, I., Seabright, A., Lai, Y.C., Tennant, D.A., Nightingale, P., Wallis, G.A., Manolopoulos, K.N., Brenner, C., Philp, A. & Lavery, G.G. 2019. Nicotinamide riboside augments the aged human skeletal muscle NAD⁺ metabolome and induces transcriptomic and anti-inflammatory signatures. Cell Reports 28(7): 1717-1728.e6.

Fleishman, J.S. & Kumar, S. 2024. Bile acid metabolism and signaling in health and disease: Molecular mechanisms and therapeutic targets. Signal Transduction and Targeted Therapy 9(1): 97.

Furuhashi, M. 2020. New insights into purine metabolism in metabolic diseases: Role of xanthine oxidoreductase activity. American Journal of Physiology-Endocrinology and Metabolism 319(5): E827-E834.

Gaillard, D., Masson, D., Garo, E., Souidi, M., Pais De Barros, J.P., Schoonjans, K., Grober, J., Besnard, P. & Thomas, C. 2021. Muricholic acids promote resistance to hypercholesterolemia in cholesterol-fed mice. International Journal of Molecular Sciences 22(13): 7163.

Guo, A., Li, K. & Xiao, Q. 2020. Fibroblast growth factor 19 alleviates palmitic acid-induced mitochondrial dysfunction and oxidative stress via the AMPK/PGC-1α pathway in skeletal muscle. Biochemical and Biophysical Research Communications 526(4): 1069-1076.

Ipata, P.L. & Pesi, R. 2018. Metabolic interaction between purine nucleotide cycle and oxypurine cycle during skeletal muscle contraction of different intensities: A biochemical reappraisal. Metabolomics 14(4): 42.

Jia, F., Liu, X. & Liu, Y. 2025. Bile acid signaling in skeletal muscle homeostasis: From molecular mechanisms to clinical applications. Frontiers in Endocrinology (Lausanne) 16(1): 1551100.

Jiang, S.Z., Wang, N.S. & Mi, S.Q. 2008. Plasma pharmacokinetics and tissue distribution of [6]-gingerol in rats. Biopharmaceutics & Drug Disposition 29(9): 529-537.

Jin, C.L., Zhang, Z.M., Song, Z.W., Gao, C.Q., Yan, H.C. & Wang, X.Q. 2020. mTORC1-mediated satellite cell differentiation is required for lysine-induced skeletal muscle growth. Journal of Agricultural and Food Chemistry 68(17): 4884-4892.

Kaiser, H., Yu, K., Pandya, C., Mendhe, B., Isales, C.M., McGee-Lawrence, M.E., Johnson, M., Fulzele, S. & Hamrick, M.W. 2019. Kynurenine, a tryptophan metabolite that increases with age, induces muscle atrophy and lipid peroxidation. Oxidative Medicine and Cellular Longevity 2019(1): 9894238.

Keitel, V., Stindt, J. & Häussinger, D. 2019. Bile acid-activated receptors: GPBAR1 (TGR5) and other G protein-coupled receptors. In Bile Acids and Their Receptors. Handbook of Experimental Pharmacology, vol. 256, edited by Fiorucci, S. & Distrutti, E. Cham: Springer. pp. 19-49.

Kudryavtseva, A.V., Krasnov, G.S., Dmitriev, A.A., Alekseev, B.Y., Kardymon, O.L., Sadritdinova, A.F., Fedorova, M.S., Pokrovsky, A.V., Melnikova, N.V., Kaprin, A.D., Moskalev, A.A. & Snezhkina, A.V. 2016. Mitochondrial dysfunction and oxidative stress in aging and cancer. Oncotarget 7(29): 44879-44905.

Lee, J., Hong, Y.S., Park, S.H. & Kang, K.Y. 2019. High serum uric acid level is associated with greater handgrip strength in the aged population. Arthritis Research & Therapy 21: 73.

Li, L.L., Cui, Y., Guo, X.H., Ma, K., Tian, P., Feng, J. & Wang, J.M. 2019. Pharmacokinetics and tissue distribution of gingerols and shogaols from ginger (Zingiber officinale Rosc.) in rats by UPLC-Q-Exactive-HRMS. Molecules 24(3): 512.

Liguori, I., Russo, G., Curcio, F., Bulli, G., Aran, L., Della-Morte, D., Gargiulo, G., Testa, G., Cacciatore, F., Bonaduce, D. & Abete, P. 2018. Oxidative stress, aging, and diseases. Clinical Interventions in Aging 13(1): 757-772.

Liu, M., Yue, Z., Zhang, B., Li, F., Liu, L. & Li, F. 2022. mTORC1 mediates the processes of lysine regulating satellite cells proliferation, apoptosis, and autophagy. Metabolites 12(9): 788.

Makpol, S., Abdul Sani, N.F., Hakimi, N.H., Ab Rani, N., Zakaria, S.N.A., Abd Rasid, A.F., Gunasekaran, G., Mohd Sahardi, N.F.N., Tan, J. K. & Abd Ghafar, N. 2020. Zingiber officinale Roscoe prevents DNA damage and improves muscle performance and bone integrity in old Sprague Dawley rats. Evidence Based Complementary Alternative Medicine 2020: 3823780.

Mohd Sahardi, N.F.N. & Makpol, S. 2019. Ginger (Zingiber officinale Roscoe) in the prevention of ageing and degenerative diseases: Review of current evidence. Evidence Based Complementary Alternative Medicine 2019: 5054395.

Mohd Sahardi, N.F.N., Jaafar, F., Tan, J.K., Mad Nordin, M.F. & Makpol, S. 2023. Elucidating the pharmacological properties of Zingiber officinale Roscoe (ginger) on muscle ageing by untargeted metabolomic profiling of human myoblasts. Nutrients 15(21): 4520.

Nile, S.H. & Park, S.W. 2015. Chromatographic analysis, antioxidant, anti-inflammatory, and xanthine oxidase inhibitory activities of ginger extracts and its reference compounds. Industrial Crops and Products 70(1): 238-244.

Packer, M. 2020. Uric acid is a biomarker of oxidative stress in the failing heart: Lessons learned from trials with allopurinol and SGLT2 inhibitors. Journal of Cardiac Failure 26(11): 977-984.

Petrocelli, J.J., Mahmassani, Z.S., Fix, D.K., Montgomery, J.A., Reidy, P.T., McKenzie, A.I., De Hart, N.M., Ferrara, P.J., Kelley, J.J., Eshima, H., Funai, K. & Drummond, M.J. 2021. Metformin and leucine increase satellite cells and collagen remodeling during disuse and recovery in aged muscle. The FASEB Journal 35(9): e21862.

Prestwich, E.G., Mangerich, A., Pang, B., Mcfaline, J.L., Lonkar, P., Sullivan, M.R., Trudel, L.J., Taghizedeh, K. & Dedon, P.C. 2013. Increased levels of inosine in a mouse model of inflammation. Chemical Research in Toxicology 26(4): 538-546.

Proungvitaya, S., Sombattheera, S., Boonsiri, P., Limpaiboon, T., Wongkham, S., Wongkham, C., Titapun, A. & Proungvitaya, T. 2017. Diagnostic value of serum bile acid composition patterns and serum glycocholic acid levels in cholangiocarcinoma. Oncology Letters 14(4): 4943-4948.

Ramprasath, T., Han, Y.M., Zhang, D., Yu, C.J. & Zou, M.H. 2021. Tryptophan catabolism and inflammation: A novel therapeutic target for aortic diseases. Frontiers in Immunology 12(1): 731701.

Razak, A.M., Zakaria, S.N.A., Abdul Sani, N.F., Ab Rani, N., Hakimi, N.H., Mohd Said, M., Tan, J.K., Gan, H.K., Mad Nordin, M.F. & Makpol, S. 2023. A subcritical water extract of soil grown Zingiber officinale Roscoe: Comparative analysis of antioxidant and anti-inflammatory effects and evaluation of bioactive metabolites. Frontiers in Pharmacology 14(1): 1006265.

Sahoo, B.M., Banik, B.K., Borah, P. & Jain, A. 2022. Reactive Oxygen Species (ROS): Key components in cancer therapies. Anti-Cancer Agents in Medicinal Chemistry 22(2): 215-222.

Sarip, M. 2012. Subcritical water extraction of 6-gingerol and 6-shogaol from Zingiber officinale. Thesis. Fakulti Kejuruteraan Kimia, Universiti Teknologi Malaysia.

Sato, T., Ito, Y. & Nagasawa, T. 2016. Regulatory effects of the L-lysine metabolites, L-2-aminoadipic acid and L-pipecolic acid, on protein turnover in C2C12 myotubes. Bioscience Biotechnology and Biochemistry 80(11): 2168-2175.

Saud Gany, S.L., Tan, J.K., Chin, K.Y., Hakimi, N.H., Ab Rani, N., Ihsan, N. & Makpol, S. 2022. Untargeted muscle tissue metabolites profiling in young, adult, and old rats supplemented with tocotrienol-rich fraction. Frontiers in Molecular Biosciences 9(1): 1008908.

Sebastiani, P., Monti, S., Lustgarten, M.S., Song, Z., Ellis, D., Tian, Q., Schwaiger-Haber, M., Stancliffe, E., Leshchyk, A., Short, M.I., Ardisson Korat, A.V., Gurinovich, A., Karagiannis, T., Li, M., Lords, H.J., Xiang, Q., Marron, M.M., Bae, H., Feitosa, M.F., Wojczynski, M.K., O'Connell, J.R., Montasser, M.E., Schupf, N., Arbeev, K., Yashin, A., Schork, N., Christensen, K., Andersen, S.L., Ferrucci, L., Rappaport, N., Perls, T.T. & Patti, G.J. 2024. Metabolite signatures of chronological age, aging, survival, and longevity. Cell Reports 43(11): 114913.

Sebih, F., Mokrane, N., Fontanel, P., Kayatekin, M., Kaabeche, M., Guiramand, J., Cohen-Solal, C., Cens, T., Rousset, M., Charnet, P., De Jésus Ferreira, M.C., Thibaud, J.B., Ménard, C., Cantel, S., Rolland, V., Vignes, M. & Roussel, J. 2021. The glutathione metabolite γ-glutamyl-glutamate partially activates glutamate NMDA receptors in central neurons with higher efficacy for GluN2B-containing receptors. Frontiers in Pharmacology 12(1): 794680.

Shang, M., Cappellesso, F., Amorim, R., Serneels, J., Virga, F., Eelen, G., Carobbio, S., Rincon, M.Y., Maechler, P., De Bock, K., Ho, P-C., Sandri, M., Ghesquière, B., Carmeliet, P., Di Matteo, M., Berardi, E. & Mazzone, M. 2020. Macrophage-derived glutamine boosts satellite cells and muscle regeneration. Nature 587(7835): 626-631.

Sies, H., Berndt, C. & Jones, D.P. 2017. Oxidative stress. Annual Review of Biochemistry 86(1): 715-748.

Songvut, P., Nakareangrit, W., Cholpraipimolrat, W., Kwangjai, J., Worasuttayangkurn, L., Watcharasit, P. & Satayavivad, J. 2024. Unraveling the interconversion pharmacokinetics and oral bioavailability of the major ginger constituents: [6]-gingerol, [6]-shogaol, and zingerone after single-dose administration in rats. Frontiers in Pharmacology 15: 1391019.

Struys, E.A. & Jakobs, C. 2010. Metabolism of lysine in α-aminoadipic semialdehyde dehydrogenase-deficient fibroblasts: Evidence for an alternative pathway of pipecolic acid formation. FEBS Letters 584(1): 181-186.

Tan, J.K., Zakaria, S.N.A., Gunasekaran, G., Abdul Sani, N.F., Nasaruddin, M.L., Jaafar, F., Abu Bakar, Z.H., Amir Hamzah, A.I.Z., Nor Aripin, K.N., Mohd Rani, M.D., Noh, N.A., Damanhuri, H.A., Mazlan, M., Makpol, S. & Wan Ngah, W.Z. 2023. Metabolomics profiling of age-associated metabolites in Malay population. Oxidative Medicine and Cellular Longevity 2023: 4416410.

Uchitomi, R., Hatazawa, Y., Senoo, N., Yoshioka, K., Fujita, M., Shimizu, T., Miura, S., Ono, Y. & Kamei, Y. 2019. Metabolomic analysis of skeletal muscle in aged mice. Scientific Reports 9(1): 10425.

Xia, J. & Wishart, D.S. 2016. Using MetaboAnalyst 3.0 for comprehensive metabolomics data analysis. Current Protocols in Bioinformatics 55(1): 14.10.11-14.10.91.

Yoshizawa, F., Mochizuki, S. & Sugahara, K. 2013. Differential dose response of mTOR signaling to oral administration of leucine in skeletal muscle and liver of rats. Bioscience, Biotechnology, and Biochemistry 77(4): 839-842.

Yuan, S. & Larsson, S.C. 2023. Epidemiology of sarcopenia: Prevalence, risk factors, and consequences. Metabolism 144(1): 155533.

Yuan, Z., Wang, J., Zhang, H., Chai, Y., Xu, Y., Miao, Y., Yuan, Z., Zhang, L., Jiang, Z. & Yu, Q. 2023. Glycocholic acid aggravates liver fibrosis by promoting the up-regulation of connective tissue growth factor in hepatocytes. Cellular Signalling 101(1): 110508.

Zhao, Q., Shen, H., Su, K-J., Tian, Q., Zhao, L-J., Qiu, C., Garrett, T.J., Liu, J., Kakhniashvili, D. & Deng, H-W. 2018. A joint analysis of metabolomic profiles associated with muscle mass and strength in Caucasian women. Aging (Albany NY) 10(10): 2624-2635.

Zheng, J., Ye, C., Hu, B., Yang, H., Yao, Q., Ma, J., Liu, Y. & Liu, H. 2021. Bile acid profiles in bile and feces of obese mice by a high-performance liquid chromatography-tandem mass spectrometry. Biotechnology and Applied Biochemistry 68(6): 1332-1341.