Molecular aspects of metformin’s anti-aging properties for muscle function and longevity in Drosophila melanogaster

Abstract

Muscle aging, characterized by the progressive loss of muscle mass and function, presents a significant clinical challenge, contributing to sarcopenia and age-related frailty. Recent research highlights metformin, a widely used anti-diabetic drug, as a promising candidate for mitigating muscle aging by targeting conserved molecular pathways. This review explores metformin’s mechanisms in Drosophila melanogaster, emphasizing its activation of AMP-activated protein kinase (AMPK) and inhibition of the mechanistic target of rapamycin (mTOR), pivotal regulators of cellular energy balance and proteostasis. Metformin enhances autophagy, reduces protein aggregation, and preserves muscle integrity by modulating autophagy-related genes, such as Atg1 and Atg8. Furthermore, the drug's suppression of ribosomal S6 kinase (S6K) and eukaryotic initiation factor 4E (eIF-4E) inhibits excessive protein synthesis, mitigating proteostatic stress. Studies in Drosophila reveal that metformin extends lifespan, reduces oxidative stress, and improves muscle function, offering insights into its translational potential for addressing sarcopenia. However, challenges remain in bridging the findings from Drosophila to humans due to species-specific differences and the need for long-term clinical studies. By elucidating the interplay of AMPK, mTOR, and autophagy pathways, this review underscores metformin’s therapeutic potential in age-related muscle decline, providing a molecular foundation for its application in geroprotective interventions. Future research should focus on optimizing dosing strategies, exploring synergistic therapies, and advancing biomarkers for muscle aging to fully harness metformin’s clinical utility in promoting healthy aging.

Keywords

Metformin / Drosophila melanogaster / Muscle Aging / AMPK / MTOR / Autophagy

Declarations

Not applicable.

Authors' contributions

Milind Umekar: Conceptualization, Supervision, Writing - Review & Editing; Mohammad Qutub: Conceptualization, Methodology, Data Curation, Writing - Original Draft, Visualization; Tanvi Premchandani: Investigation, Visualization, Writing - Review & Editing; Amol Tatode: Supervision, Project Administration, Methodology, Writing - Review & Editing; Jayshree Taksande: Writing - Review & Editing; Priyanka Singanwad: Visualization; Mayur Kale: Investigation, Formal Analysis; Mithun Maniyar: Writing - Review & Editing; Ujban Md Hussain: Writing - Review & Editing.

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Availability of data and materials

Not applicable.

Funding

Not applicable.

Declaration of Competing Interest

Not applicable.

Acknowledgements

Not applicable.

Authors' other information

Not applicable.

References

[1] Aman Y, Schmauck-Medina T, Hansen M, et al. Autophagy in healthy aging and disease. Nat Aging. 2021; 1(8):634-650. https://doi.org/10.1038/s43587-021-00098-4

[2] Slade L, Etheridge T, Szewczyk NJ. Consolidating multiple evolutionary theories of ageing suggests a need for new approaches to study genetic contributions to ageing decline. Ageing Res Rev. 2024; 100:102456. https://doi.org/10.1016/j.arr.2024.102456

[3] Larsson L, Degens H, Li M, et al. Sarcopenia: aging-related loss of muscle mass and function. Physiol Rev. 2019; 99(1):427-511. https://doi.org/10.1152/physrev.00061.2017

[4] Boccardi V. Sarcopenia: a dive into metabolism to promote a multimodal, preventive, and regenerative approach. Mech Ageing Dev. 2024; 219:111941. https://doi.org/10.1016/j.mad.2024.111941

[5] Ardeljan AD, Hurezeanu R. Sarcopenia StatPearls. 2024.

[6] Madreiter-Sokolowski CT, Hiden U, Krstic J, et al. Targeting organ-specific mitochondrial dysfunction to improve biological aging. Pharmacol Ther. 2024; 262:108710. https://doi.org/10.1016/j.pharmthera.2024.108710

[7] Trigo D, Nadais A, Carvalho A, et al. Mitochondria dysfunction and impaired response to oxidative stress promotes proteostasis disruption in aged human cells. Mitochondrion. 2023; 69:1-9. https://doi.org/10.1016/j.mito.2022.10.002

[8] Li X, Jiang O, Chen M, Wang S. Mitochondrial homeostasis: shaping health and disease. Curr Med. 2024; 3(1):5. https://doi.org/10.1007/s44194-024-00032-x

[9] El-Sebaie M, Elwakil W. Biomarkers of sarcopenia: an unmet need. Egypt Rheumatol Rehabil. 2023; 50(1):45. https://doi.org/10.1186/s43166-023-00213-w

[10] Wiedmer P, Jung T, Castro JP, et al. Sarcopenia - molecular mechanisms and open questions. Ageing Res Rev. 2021; 65:101200. https://doi.org/10.1016/j.arr.2020.101200

[11] Giorgi C, Marchi S, Simoes ICM, Ren Z, Morciano G, Perrone M, Patalas-Krawczyk P, Borchard S, Jędrak P, Pierzynowska K, Szymański J, Wang DQ, Portincasa P, Węgrzyn G, Zischka H, Dobrzyn P, Bonora M, Duszynski J, Rimessi A,... Wieckowski MR. Mitochondria React Oxyg Species Aging AgeRelat Dis. 2018:209-344. https://doi.org/10.1016/bs.ircmb.2018.05.006

[12] Lin P, Lu Y, Zheng J, Lin Y, Zhao X, Cui L. Strategic disruption of cancer’s powerhouse: precise nanomedicine targeting of mitochondrial metabolism. J Nanobiotechnol. 2024; 22(1):318. https://doi.org/10.1186/s12951-024-02585-3

[13] Antuña E, Cachán-Vega C, Bermejo-Millo JC, et al. Inflammaging: implications in sarcopenia. Int J Mol Sci. 2022; 23(23):15039. https://doi.org/10.3390/ijms232315039

[14] Damanti S, Senini E, De Lorenzo R, et al. Acute sarcopenia: mechanisms and management. Nutrients. 2024; 16(20):3428. https://doi.org/10.3390/nu16203428

[15] Sahinyan K, Lazure F, Blackburn DM, Soleimani VD. Decline of regenerative potential of old muscle stem cells: contribution to muscle aging. FEBS J. 2023; 290(5):1267-1289. https://doi.org/10.1111/febs.16352

[16] Tseng C-S, Chao Y-W, Liu Y-H, Huang Y-S, Chao H-W. Dysregulated proteostasis network in neuronal diseases. Front Cell Dev Biol. 2023; 11. https://doi.org/10.3389/fcell.2023.1075215

[17] Henze H, Jung MJ, Ahrens HE, Steiner S, von Maltzahn J. Skeletal muscle aging - stem cells in the spotlight. Mech Ageing Dev. 2020; 189:111283. https://doi.org/10.1016/j.mad.2020.111283

[18] Chinvattanachot G, Rivas D, Duque G. Mechanisms of muscle cells alterations and regeneration decline during aging. Ageing Res Rev. 2024; 102:102589. https://doi.org/10.1016/j.arr.2024.102589

[19] Huo F, Liu Q, Liu H. Contribution of muscle satellite cells to sarcopenia. Front Physiol. 2022; 13. https://doi.org/10.3389/fphys.2022.892749

[20] Vainshtein A, Sandri M. Signaling pathways that control muscle mass. Int J Mol Sci. 2020; 21(13):4759. https://doi.org/10.3390/ijms21134759

[21] Yoshida T, Delafontaine P. Mechanisms of IGF-1-mediated regulation of skeletal muscle hypertrophy and atrophy. Cells. 2020; 9(9):1970. https://doi.org/10.3390/cells9091970

[22] Simcox J, Lamming DW. The central moTOR of metabolism. Dev Cell. 2022; 57(6):691-706. https://doi.org/10.1016/j.devcel.2022.02.024

[23] Sartori R, Romanello V, Sandri M. Mechanisms of muscle atrophy and hypertrophy: implications in health and disease. Nat Commun. 2021; 12(1):330. https://doi.org/10.1038/s41467-020-20123-1

[24] Dönertaş HM, Fuentealba M, Partridge L, Thornton JM. Identifying potential ageing-modulating drugs in silico. Trends Endocrinol Metab. 2019; 30(2):118-131. https://doi.org/10.1016/j.tem.2018.11.005

[25] Kennedy BK, Pennypacker JK. Drugs that modulate aging: the promising yet difficult path ahead. Transl Res. 2014; 163(5):456-465. https://doi.org/10.1016/j.trsl.2013.11.007

[26] Drzewoski J, Hanefeld M. The current and potential therapeutic use of Metformin—The good old drug. Pharmaceuticals. 2021; 14(2):122. https://doi.org/10.3390/ph14020122

[27] Soukas AA, Hao H, Wu L. Metformin as anti-aging therapy: is it for everyone? Trends Endocrinol Metab. 2019; 30(10):745-755. https://doi.org/10.1016/j.tem.2019.07.015

[28] Lv Z, Guo Y. Metformin and its benefits for various diseases. Front Endocrinol. 2020; 11. https://doi.org/10.3389/fendo.2020.00191

[29] Herman R, Kravos NA, Jensterle M, Janež A, Dolžan V. Metformin and insulin resistance: a review of the underlying mechanisms behind changes in GLUT4-Mediated glucose transport. Int J Mol Sci. 2022; 23(3):1264. https://doi.org/10.3390/ijms23031264

[30] Rodríguez C, Muñoz M, Contreras C, Prieto D. AMPK, metabolism, and vascular function. FEBS J. 2021; 288(12):3746-3771. https://doi.org/10.1111/febs.15863

[31] Xu Y, Bai L, Yang X, et al. Recent advances in anti-inflammation via AMPK activation. Heliyon. 2024; 10(13):e33670. https://doi.org/10.1016/j.heliyon.2024.e33670

[32] Kulkarni AS, Gubbi S, Barzilai N. Benefits of metformin in attenuating the hallmarks of aging. Cell Metab. 2020; 32(1):15-30. https://doi.org/10.1016/j.cmet.2020.04.001

[33] Baenas N, Wagner AE. Drosophila melanogaster as an alternative model organism in nutrigenomics. Genes Nutr. 2019; 14(1):14. https://doi.org/10.1186/s12263-019-0641-y

[34] Qutub M, Tatode A, Taksande J, et al. Stimuli-responsive supramolecular hydrogels for paclitaxel delivery: progress and prospects. Asp Mol Med. 2025; 5:100062. https://doi.org/10.1016/j.amolm.2024.100062

[35] Baenas N, Wagner AE. Drosophila melanogaster as a model organism for obesity and Type-2 diabetes mellitus by applying High-Sugar and High-Fat diets. Biomolecules. 2022; 12(2):307. https://doi.org/10.3390/biom12020307

[36] Ans K, Lüersen K, von Frieling J, Roeder T, Rimbach G. Dietary lithium stimulates female fecundity in drosophila melanogaster. BioFactors. 2024; 50(2):326-346. https://doi.org/10.1002/biof.2007

[37] Berk Ş, Cetin A, Özdemir Ö. Ü Pektaş AN, Yurtcu N, Dastan SD. The combination of metformin and high glucose increased longevity of caenorhabditis elegans a DAF-16/FOXO-independent manner: cancer/diabetic model via C. Elegans. Front Endocrinol. 2024; 15:1435098. https://doi.org/10.3389/FENDO.2024.1435098/BIBTEX

[38] De Haes W, Frooninckx L, Van Assche R, et al. Metformin promotes lifespan through mitohormesis via the peroxiredoxin PRDX-2. Proc Natl Acad Sci. 2014; 111(24) https://doi.org/10.1073/pnas.1321776111

[39] Hu D, Xie F, Xiao Y, et al. Metformin: a potential candidate for targeting aging mechanisms. Aging Dis. 2021; 12(2):480. https://doi.org/10.14336/AD.2020.0702

[40] Wang Y, An H, Liu T, et al. Metformin improves mitochondrial respiratory activity through activation of AMPK. Cell Rep. 2019; 29(6):1511-1523.e5. https://doi.org/10.1016/j.celrep.2019.09.070

[41] Szymczak-Pajor I, Wenclewska S, Śliwińska A. Metabolic action of metformin. Pharmaceuticals. 2022; 15(7):810. https://doi.org/10.3390/ph15070810

[42] Entezari M, Hashemi D, Taheriazam A, et al. AMPK signaling in diabetes mellitus, insulin resistance and diabetic complications: a pre-clinical and clinical investigation. Biomed Pharmacother. 2022; 146:112563. https://doi.org/10.1016/j.biopha.2021.112563

[43] LaMoia TE, Shulman GI. Cellular and molecular mechanisms of metformin action. Endocr Rev. 2021; 42(1):77-96. https://doi.org/10.1210/endrev/bnaa023

[44] Herman R, Kravos NA, Jensterle M, Janež A, Dolžan V. Metformin and insulin resistance: a review of the underlying mechanisms behind changes in GLUT4-Mediated glucose transport. Int J Mol Sci. 2022; 23(3):1264. https://doi.org/10.3390/ijms23031264

[45] Salminen A, Kaarniranta K. AMP-activated protein kinase (AMPK) controls the aging process via an integrated signaling network. Ageing Res Rev. 2012; 11(2):230-241. https://doi.org/10.1016/j.arr.2011.12.005

[46] Sun Y, Wang H, Qu T, et al. mTORC2: a multifaceted regulator of autophagy. Cell Commun Signal. 2023; 21(1):4. https://doi.org/10.1186/s12964-022-00859-7

[47] Marafie SK, Al-Mulla F, Abubaker J. mTOR: its critical role in metabolic diseases, cancer, and the aging process. Int J Mol Sci. 2024; 25(11):6141. https://doi.org/10.3390/ijms25116141

[48] Mannick JB, Lamming DW. Targeting the biology of aging with mTOR inhibitors. Nat Aging. 2023; 3(6):642-660. https://doi.org/10.1038/s43587-023-00416-y

[49] Khalil MI, Ali MM, Holail J, Houssein M. Growth or death? Control of cell destiny by mTOR and autophagy pathways. Prog Biophys Mol Biol. 2023; 185:39-55. https://doi.org/10.1016/j.pbiomolbio.2023.10.002

[50] Sciarretta S, Forte M, Frati G, Sadoshima J. New insights into the role of mTOR signaling in the cardiovascular system. Circ Res. 2018; 122(3):489-505. https://doi.org/10.1161/CIRCRESAHA.117.311147

[51] Cheng F-F, Liu Y-L, Du J, Lin J-T. Metformin’s mechanisms in attenuating hallmarks of aging and Age-Related disease. Aging Dis. 2022; 13(4):970. https://doi.org/10.14336/AD.2021.1213

[52] Lee S-H, Min K-J. Drosophila melanogaster as a model system in the study of pharmacological interventions in aging. Transl Med Aging. 2019; 3:98-103. https://doi.org/10.1016/j.tma.2019.09.004

[53] Liu J-K. Antiaging agents: safe interventions to slow aging and healthy life span extension. Nat Prod Bioprospecting. 2022; 12(1):18. https://doi.org/10.1007/s13659-022-00339-y

[54] Aliper A, Jellen L, Cortese F, et al. Towards natural mimetics of metformin and rapamycin. Aging. 2017; 9(11):2245-2268. https://doi.org/10.18632/aging.101319

[55] Rand MD, Tennessen JM, Mackay TFC, Anholt RRH. Perspectives on the drosophila melanogaster model for advances in toxicological science. Curr Protoc. 2023; 3(8) https://doi.org/10.1002/cpz1.870

[56] Fischer FP, Karge RA, Weber YG, Koch H, Wolking S, Voigt A. Drosophila melanogaster as a versatile model organism to study genetic epilepsies: an overview. Front Mol Neurosci. 2023; 16. https://doi.org/10.3389/fnmol.2023.1116000

[57] Asthana J, Shravage BV. Exploring therapeutic potential of mitophagy modulators using drosophila models of Parkinson’s disease. Front Aging Neurosci. 2022; 14. https://doi.org/10.3389/fnagi.2022.986849

[58] Zheng X. An introductory guide to using Bloomington drosophila stock center and FlyBase for aging research. Cells. 2024; 13(14):1192. https://doi.org/10.3390/cells13141192

[59] Tsurumi A, Li WX. Aging mechanisms—A perspective mostly from drosophila. Adv Genet. 2020; 1(1) https://doi.org/10.1002/ggn2.10026

[60] Cassidy D, Epiney DG, Salameh C, et al. Evidence for premature aging in a drosophila model of werner syndrome. Exp Gerontol. 2019; 127:110733. https://doi.org/10.1016/j.exger.2019.110733

[61] Maruzs T, Simon-Vecsei Z, Kiss V, Csizmadia T, Juhász G. On the Fly: recent progress on autophagy and aging in drosophila. Front Cell Dev Biol. 2019; 7. https://doi.org/10.3389/fcell.2019.00140

[62] Demir E, Kacew S. Drosophila as a robust model system for assessing autophagy: a review. Toxics. 2023; 11(8):682. https://doi.org/10.3390/toxics11080682

[63] Aparicio R, Rana A, Walker DW. Upregulation of the autophagy adaptor p62/SQSTM1 prolongs health and lifespan in Middle-Aged drosophila. Cell Rep. 2019; 28(4):1029-1040.e5. https://doi.org/10.1016/j.celrep.2019.06.070

[64] Li P, Ma Y, Yu C, et al. Autophagy and aging: roles in skeletal muscle, eye, brain and hepatic tissue. Front Cell Dev Biol. 2021; 9. https://doi.org/10.3389/fcell.2021.752962

[65] Hu D, Xie F, Xiao Y, et al. Metformin: a potential candidate for targeting aging mechanisms. Aging Dis. 2021; 12(2):480. https://doi.org/10.14336/AD.2020.0702

[66] Le Pelletier L, Mantecon M, Gorwood J, et al. Metformin alleviates stress-induced cellular senescence of aging human adipose stromal cells and the ensuing adipocyte dysfunction. ELife. 2021; 10. https://doi.org/10.7554/eLife.62635

[67] Fernando R, Drescher C, Nowotny K, Grune T, Castro JP. Impaired proteostasis during skeletal muscle aging. Free Radic Biol Med. 2019; 132:58-66. https://doi.org/10.1016/j.freeradbiomed.2018.08.037

[68] Vilchez D, Saez I, Dillin A. The role of protein clearance mechanisms in organismal ageing and age-related diseases. Nat Commun. 2014; 5(1):5659. https://doi.org/10.1038/ncomms6659

[69] Vilchez D, Saez I, Dillin A. The role of protein clearance mechanisms in organismal ageing and age-related diseases. Nat Commun. 2014; 5(1):5659. https://doi.org/10.1038/ncomms6659

[70] Barmaki H, Nourazarian A, Khaki-Khatibi F. Proteostasis and neurodegeneration: a closer look at autophagy in Alzheimer’s disease. Front Aging Neurosci. 2023; 15. https://doi.org/10.3389/fnagi.2023.1281338

[71] Rey V, Tamargo-Gómez I. From kinases to diseases: investigating the role of AMPK in human pathologies. Kinases Phosphatases. 2023; 1(3):181-205. https://doi.org/10.3390/kinasesphosphatases1030012

[72] Cokorinos EC, Delmore J, Reyes AR, et al. Activation of skeletal muscle AMPK promotes glucose disposal and glucose lowering in Non-human primates and mice. Cell Metab. 2017; 25(5):1147-1159.e10. https://doi.org/10.1016/j.cmet.2017.04.010

[73] Zhang Y, Zong H, Qi Y, et al. Anxiolytic effect of antidiabetic metformin is mediated by AMPK activation in mPFC inhibitory neurons. Mol Psychiatry. 2023; 28(9):3955-3965. https://doi.org/10.1038/s41380-023-02283-w

[74] Hardie DG. AMP-activated protein kinase—an energy sensor that regulates all aspects of cell function. Genes Dev. 2011; 25(18):1895-1908. https://doi.org/10.1101/gad.17420111

[75] Steinberg GR, Carling D. AMP-activated protein kinase: the current landscape for drug development. Nat Rev Drug Discov. 2019; 18(7):527-551. https://doi.org/10.1038/s41573-019-0019-2

[76] Meng S, Cao J, He Q, et al. Metformin activates AMP-activated protein kinase by promoting formation of the αβγ heterotrimeric complex. J Biol Chem. 2015; 290(6):3793-3802. https://doi.org/10.1074/jbc.M114.604421

[77] Hardie DG. AMP-activated protein kinase—an energy sensor that regulates all aspects of cell function. Genes Dev. 2011; 25(18):1895-1908. https://doi.org/10.1101/gad.17420111

[78] Sharma A, Anand SK, Singh N, Dwivedi UN, Kakkar P. AMP-activated protein kinase: an energy sensor and survival mechanism in the reinstatement of metabolic homeostasis. Exp Cell Res. 2023; 428(1):113614. https://doi.org/10.1016/j.yexcr.2023.113614

[79] Min SH, Song DK, Lee CH, Roh E, Kim M-S. Hypothalamic AMP-Activated protein kinase as a Whole-Body energy sensor and regulator. Endocrinol Metab. 2024; 39(1):1-11. https://doi.org/10.3803/EnM.2024.1922

[80] Garcia D, Shaw RJ. AMPK: mechanisms of cellular energy sensing and restoration of metabolic balance. Mol Cell. 2017; 66(6):789-800. https://doi.org/10.1016/j.molcel.2017.05.032

[81] Yi D, Yu H, Lu K, et al. AMPK signaling in energy control, cartilage biology, and osteoarthritis. Front Cell Dev Biol. 2021; 9. https://doi.org/10.3389/fcell.2021.696602

[82] Mihaylova MM, Shaw RJ. The AMPK signalling pathway coordinates cell growth, autophagy and metabolism. Nat Cell Biol. 2011; 13(9):1016-1023. https://doi.org/10.1038/ncb2329

[83] Foretz M, Guigas B, Viollet B. Metformin: update on mechanisms of action and repurposing potential. Nat Rev Endocrinol. 2023; 19(8):460-476. https://doi.org/10.1038/s41574-023-00833-4

[84] Zhou G, Myers R, Li Y, et al. Role of AMP-activated protein kinase in mechanism of metformin action. J Clin Investig. 2001; 108(8):1167-1174. https://doi.org/10.1172/JCI13505

[85] Ma T, Tian X, Zhang B, Li M, Wang Y, Yang C, Wu J, Wei X, Qu Q, Yu Y, Long S, Feng J-W, Li C, Zhang C, Xie C, Wu Y, Xu Z, Chen J, Yu Y,... Lin S-C. Low-dose metformin targets the lysosomal AMPK pathway through PEN2. Nature. 2022; 603(7899):159-165. https://doi.org/10.1038/s41586-022-04431-8

[86] Laws KM, Drummond-Barbosa D. AMP-activated protein kinase has diet-dependent and -independent roles in drosophila oogenesis. Dev Biol. 2016; 420(1):90-99. https://doi.org/10.1016/j.ydbio.2016.10.006

[87] Stancu AL. AMPK activation can delay aging. Discoveries. 2015; 3(4):e53. https://doi.org/10.15190/d.2015.45

[88] Ashraf N, Van Nostrand JL. Fine-tuning AMPK in physiology and disease using point-mutant mouse models. Dis Models Mech. 2024; 17(8) https://doi.org/10.1242/dmm.050798

[89] Foretz M, Guigas B, Viollet B. Metformin: update on mechanisms of action and repurposing potential. Nat Rev Endocrinol. 2023; 19(8):460-476. https://doi.org/10.1038/s41574-023-00833-4

[90] Zhang Y, Guo R, Wang S-S, et al. Autophagy-related proteins in genome stability: Autophagy-Dependent and independent actions. Int J Biol Sci. 2022; 18(14):5329-5344. https://doi.org/10.7150/ijbs.76134

[91] Salminen A. AMPK signaling inhibits the differentiation of myofibroblasts: impact on age-related tissue fibrosis and degeneration. Biogerontology. 2024; 25(1):83-106. https://doi.org/10.1007/s10522-023-10072-9

[92] Galal MA, Al-Rimawi M, Hajeer A, Dahman H, Alouch S, Aljada A. Metformin: a Dual-Role player in cancer treatment and prevention. Int J Mol Sci. 2024; 25(7):4083. https://doi.org/10.3390/ijms25074083

[93] Wang C, Chen B, Feng Q, Nie C, Li T. Clinical perspectives and concerns of metformin as an anti-aging drug. AGING Med. 2020; 3(4):266-275. https://doi.org/10.1002/agm2.12135

[94] Song Y, Wu Z, Zhao P. The function of metformin in Aging-Related musculoskeletal disorders. Front Pharmacol. 2022; 13. https://doi.org/10.3389/fphar.2022.865524

[95] Mirzoyan Z., Valenza A., Zola S., Bonfanti C., Arnaboldi L., Ferrari N., Pollard J., Lupi V., Cassinelli M., Frattaroli M., Sahin M., Pasini M.E., & Bellosta P. (2023). A Novel Drosophila Model to Investigate Adipose Tissue Macrophage Infiltration (ATM) and Obesity highlights the Therapeutic Potential of Attenuating Eiger/TNF α Signaling to Ameliorate Insulin Resistance and ATM. 1-30. https://doi.org/10.1101/2023.07.06.548016.

[96] Mirzoyan Z, Valenza A, Zola S, et al. A drosophila model targets Eiger/TNFα to alleviate obesity-related insulin resistance and macrophage infiltration. DMM Dis Models Mech. 2023; 16(11):1-30. https://doi.org/10.1242/dmm.050388

[97] Morita M, Gravel S-P, Hulea L, et al. mTOR coordinates protein synthesis, mitochondrial activity and proliferation. Cell Cycle. 2015; 14(4):473-480. https://doi.org/10.4161/15384101.2014.991572

[98] Zou Z, Tao T, Li H, Zhu X. mTOR signaling pathway and mTOR inhibitors in cancer: progress and challenges. Cell Biosci. 2020; 10(1):31. https://doi.org/10.1186/s13578-020-00396-1

[99] Howell JJ, Hellberg K, Turner M, et al. Metformin inhibits hepatic mTORC1 signaling via Dose-Dependent mechanisms involving AMPK and the TSC complex. Cell Metab. 2017; 25(2):463-471. https://doi.org/10.1016/j.cmet.2016.12.009

[100] Amin S, Lux A, O’Callaghan F. The journey of metformin from glycaemic control to mTOR inhibition and the suppression of tumour growth. Br J Clin Pharmacol. 2019; 85(1):37-46. https://doi.org/10.1111/bcp.13780

[101] Smiles WJ, Ovens AJ, Kemp BE, Galic S, Petersen J, Oakhill JS. New developments in AMPK and mTORC1 cross-talk. Essays Biochem. 2024. https://doi.org/10.1042/EBC20240007

[102] Amin S, Lux A, O’Callaghan F. The journey of metformin from glycaemic control to mTOR inhibition and the suppression of tumour growth. Br J Clin Pharmacol. 2019; 85(1):37-46. https://doi.org/10.1111/bcp.13780

[103] Yang L, Miao L, Liang F, et al. The mTORC1 effectors S6K1 and 4E-BP play different roles in CNS axon regeneration. Nat Commun. 2014; 5(1):5416. https://doi.org/10.1038/ncomms6416

[104] Magnuson B, Ekim B, Fingar DC. Regulation and function of ribosomal protein S 6 kinase (S6K) within mTOR signalling networks. Biochem J. 2012; 441(1):1-21. https://doi.org/10.1042/BJ20110892

[105] Kezic A, Popovic L, Lalic K. mTOR inhibitor therapy and metabolic consequences: where do we stand? Oxid Med Cell Longev. 2018; 2018(1) https://doi.org/10.1155/2018/2640342

[106] Tang J-X, Xiao F-H. Editorial: the regulation of proteostasis in aging. Front Cell Dev Biol. 2023; 11. https://doi.org/10.3389/fcell.2023.1221510

[107] Ibarra-Lecue I, Diez-Alarcia R, Morentin B, Meana JJ, Callado LF, Urigüen L. Ribosomal protein S 6 hypofunction in postmortem human brain links mTORC1-Dependent signaling and schizophrenia. Front Pharmacol. 2020;11. https://doi.org/10.3389/fphar.2020.00344

[108] Wu X, Xie W, Xie W, Wei W, Guo J. Beyond controlling cell size: functional analyses of S6K in tumorigenesis. Cell Death Dis. 2022; 13(7):646. https://doi.org/10.1038/s41419-022-05081-4

[109] Zhang P, Catterson JH, Grönke S, Partridge L. Inhibition of S6K lowers age-related inflammation and increases lifespan through the endolysosomal system. Nat Aging. 2024; 4(4):491-509. https://doi.org/10.1038/s43587-024-00578-3

[110] Petersen MC, Shulman GI. Mechanisms of insulin action and insulin resistance. Physiol Rev. 2018; 98(4):2133-2223. https://doi.org/10.1152/physrev.00063.2017

[111] Fagerberg L, Hallström BM, Oksvold P, et al. Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics. Mol Cell Proteom MCP. 2014; 13(2):397-406. https://doi.org/10.1074/mcp.M113.035600

[112] Roiuk M, Neff M, Teleman AA. eIF4E-independent translation is largely eIF3d-dependent. Nat Commun. 2024; 15(1):6692. https://doi.org/10.1038/s41467-024-51027-z

[113] Howell JJ, Hellberg K, Turner M, et al. Metformin inhibits hepatic mTORC1 signaling via Dose-Dependent mechanisms involving AMPK and the TSC complex. Cell Metab. 2017; 25(2):463-471. https://doi.org/10.1016/j.cmet.2016.12.009

[114] Yang M, Lu Y, Piao W, Jin H. The translational regulation in mTOR pathway. Biomolecules. 2022; 12(6):802. https://doi.org/10.3390/biom12060802

[115] Morita M, Gravel S-P, Hulea L, et al. mTOR coordinates protein synthesis, mitochondrial activity and proliferation. Cell Cycle. 2015; 14(4):473-480. https://doi.org/10.4161/15384101.2014.991572

[116] Mannick JB, Lamming DW. Targeting the biology of aging with mTOR inhibitors. Nat Aging. 2023; 3(6):642-660. https://doi.org/10.1038/s43587-023-00416-y

[117] Querfurth H, Lee H-K. Mammalian/mechanistic target of rapamycin (mTOR) complexes in neurodegeneration. Mol Neurodegener. 2021; 16(1):44. https://doi.org/10.1186/s13024-021-00428-5

[118] Howell JJ, Hellberg K, Turner M, et al. Metformin inhibits hepatic mTORC1 signaling via Dose-Dependent mechanisms involving AMPK and the TSC complex. Cell Metab. 2017; 25(2):463-471. https://doi.org/10.1016/j.cmet.2016.12.009

[119] Agius L, Ford BE, Chachra SS. The metformin mechanism on gluconeogenesis and AMPK activation: the metabolite perspective. Int J Mol Sci. 2020; 21(9):3240. https://doi.org/10.3390/ijms21093240

[120] Chu J, Pelletier J. Therapeutic opportunities in eukaryotic translation. Cold Spring Harb Perspect Biol. 2018; 10(6):a032995. https://doi.org/10.1101/cshperspect.a032995

[121] Nieto-Torres JL, Hansen M. Macroautophagy and aging: the impact of cellular recycling on health and longevity. Mol Asp Med. 2021; 82:101020. https://doi.org/10.1016/j.mam.2021.101020

[122] Ryter SW, Cloonan SM, Choi AMK. Autophagy: a critical regulator of cellular metabolism and homeostasis. Mol Cells. 2013; 36(1):7-16. https://doi.org/10.1007/s10059-013-0140-8

[123] Gómez-Virgilio L, Silva-Lucero M-C, Flores-Morelos D-S, et al. Autophagy: a key regulator of homeostasis and disease: an overview of molecular mechanisms and modulators. Cells. 2022; 11(15):2262. https://doi.org/10.3390/cells11152262

[124] Zhao YG, Codogno P, Zhang H. Machinery, regulation and pathophysiological implications of autophagosome maturation. Nat Rev Mol Cell Biol. 2021; 22(11):733-750. https://doi.org/10.1038/s41580-021-00392-4

[125] Misirkic Marjanovic MS, Vucicevic LM, Despotovic AR, Stamenkovic MM, Janjetovic KD. Dual anticancer role of metformin: an old drug regulating AMPK dependent/independent pathways in metabolic, oncogenic/tumorsuppresing and immunity context. Am J Cancer Res. 2021; 11(11):5625-5643 https://pubmed.ncbi.nlm.nih.gov/34873484.

[126] Sadria M, Layton AT. Interactions among mTORC, AMPK and SIRT: a computational model for cell energy balance and metabolism. Cell Commun Signal. 2021; 19(1):57. https://doi.org/10.1186/s12964-021-00706-1

[127] Rabanal-Ruiz Y, Otten EG, Korolchuk VI. mTORC1 as the main gateway to autophagy. Essays Biochem. 2017; 61(6):565-584. https://doi.org/10.1042/EBC20170027

[128] Alers S, Löffler AS, Wesselborg S, Stork B. Role of AMPK-mTOR-Ulk1/2 in the regulation of autophagy: cross talk, shortcuts, and feedbacks. Mol Cell Biol. 2012; 32(1):2-11. https://doi.org/10.1128/MCB.06159-11

[129] Lim JJ, Hyun S. Minocycline treatment improves proteostasis during drosophila aging via autophagy mediated by FOXO and Hsp70. Biomed Pharmacother. 2022; 149:112803. https://doi.org/10.1016/j.biopha.2022.112803

[130] Sehrawat A, Mishra J, Mastana SS, et al. Dysregulated autophagy: a key player in the pathophysiology of type 2 diabetes and its complications. Biochim Et Biophys Acta (BBA) Mol Basis Dis. 2023; 1869(4):166666. https://doi.org/10.1016/j.bbadis.2023.166666

[131] Gonzalez CD, Lee M-S, Marchetti P, et al. The emerging role of autophagy in the pathophysiology of diabetes mellitus. Autophagy. 2011; 7(1):2-11. https://doi.org/10.4161/auto.7.1.13044

[132] Tabibzadeh S. Role of autophagy in aging: the good, the bad, and the ugly. Aging Cell. 2023; 22(1) https://doi.org/10.1111/acel.13753

[133] Ge X, Wang L, Fei A, Ye S, Zhang Q. Research progress on the relationship between autophagy and chronic complications of diabetes. Front Physiol. 2022; 13. https://doi.org/10.3389/fphys.2022.956344

[134] Nguyen HT, Wiederkehr A, Wollheim CB, Park K-S. Regulation of autophagy by perilysosomal calcium: a new player in β-cell lipotoxicity. Exp Mol Med. 2024; 56(2):273-288. https://doi.org/10.1038/s12276-024-01161-x

[135] Xie G, Jin H, Mikhail H, et al. Autophagy in sarcopenia: possible mechanisms and novel therapies. Biomed Pharmacother. 2023; 165:115147. https://doi.org/10.1016/j.biopha.2023.115147

[136] Vainshtein A, Sandri M. Signaling pathways that control muscle mass. Int J Mol Sci. 2020; 21(13):4759. https://doi.org/10.3390/ijms21134759

[137] Sebastián D, Zorzano A. Self-Eating for muscle fitness: autophagy in the control of energy metabolism. Dev Cell. 2020; 54(2):268-281. https://doi.org/10.1016/j.devcel.2020.06.030

[138] Querfurth H, Lee H-K. Mammalian/mechanistic target of rapamycin (mTOR) complexes in neurodegeneration. Mol Neurodegener. 2021; 16(1):44. https://doi.org/10.1186/s13024-021-00428-5

[139] Mao Z, Zhang W. Role of mTOR in glucose and lipid metabolism. Int J Mol Sci. 2018; 19(7):2043. https://doi.org/10.3390/ijms19072043

[140] Portier I, Manne BK, Kosaka Y, et al. Aging-related alterations in mechanistic target of rapamycin signaling promote platelet hyperreactivity and thrombosis. J Thromb Haemost. 2024; 22(9):2576-2588. https://doi.org/10.1016/j.jtha.2024.05.025

[141] Panwar V, Singh A, Bhatt M, et al. Multifaceted role of mTOR (mammalian target of rapamycin) signaling pathway in human health and disease. Signal Transduct Target Ther. 2023; 8(1) https://doi.org/10.1038/s41392-023-01608-z

[142] Milanović M, Bekić M, Đokić J, Vučević D, Čolić M, Tomić S. Exogenous α-ketoglutarate modulates redox metabolism and functions of human dendritic cells, altering their capacity to polarise t cell response. Int J Biol Sci. 2024; 20(3):1064-1087. https://doi.org/10.7150/IJBS.91109

[143] Bayliak MM, Gospodaryov DV, Lushchak VI. Mimicking caloric restriction for anti-aging effects: the pro-oxidant role of alpha-ketoglutarate. Curr Opin Toxicol. 2022; 30:100339. https://doi.org/10.1016/J.COTOX.2022.02.012

[144] Nie L, He K, Qiu C, et al. Tetramethylpyrazine nitrone alleviates D-galactose-induced murine skeletal muscle aging and motor deficits by activating the AMPK signaling pathway. Biomed Pharmacother. 2024; 173. https://doi.org/10.1016/j.biopha.2024.116415

[145] Wang T, Jing M, Zhang T, Zhang Z, Sun Y, Wang Y. Tetramethylpyrazine nitrone TBN extends the lifespan of C. Elegans by activating the Nrf2/SKN-1 signaling pathway. Biochem Biophys Res Commun. 2022; 614:107-113. https://doi.org/10.1016/J.BBRC.2022.05.009

[146] Luo X, Liu P, Ye X, et al. Curcumin improves atrial fibrillation susceptibility by regulating tsRNA expression in aging mouse atrium. PeerJ. 2024; 12(7):e17495. https://doi.org/10.7717/peerj.17495

[147] Bisht A, Tewari D, Kumar S, Chandra S. Network pharmacology, molecular docking, and molecular dynamics simulation to elucidate the mechanism of anti-aging action of tinospora cordifolia. Mol Divers. 2024; 28(3):1743-1763. https://doi.org/10.1007/s11030-023-10684-w

[148] You R, Shi L, Chen S, et al. Fabrication of Resveratrol-Loaded scaffolds and their application for delaying cell senescence in vitro. Macromol Biosci. 2022; 22(3) https://doi.org/10.1002/mabi.202100440

[149] Liang QX, Lin YH, Zhang CH, et al. Resveratrol increases resistance of mouse oocytes to postovulatory aging in vivo. Aging. 2018; 10(7):1586-1596. https://doi.org/10.18632/aging.101494

[150] Madeo F, Eisenberg T, Pietrocola F, Kroemer G. Spermidine in health and disease. Science. 2018; 359(6374) https://doi.org/10.1126/SCIENCE.AAN2788

[151] Santín-Márquez R, Alarcón-Aguilar A, López-Diazguerrero NE, Chondrogianni N, Königsberg M. Sulforaphane - role in aging and neurodegeneration. GeroScience. 2019; 41(5):655-670. https://doi.org/10.1007/S11357-019-00061-7

[152] Liu L, Guo P, Wang P, Zheng S, Qu Z, Liu N. The review of Anti-aging mechanism of polyphenols on caenorhabditis elegans. Front Bioeng Biotechnol. 2021; 9. https://doi.org/10.3389/FBIOE.2021.635768/FULL

[153] Chen Y, Onken B, Chen H, et al. Mechanism of longevity extension of caenorhabditis elegans induced by pentagalloyl glucose isolated from eucalyptus leaves. J Agric Food Chem. 2014; 62(15):3422-3431. https://doi.org/10.1021/JF500210P

[154] Thanapairoje K, Junsiritrakhoon S, Wichaiyo S, Osman MA, Supharattanasitthi W. Anti-ageing effects of FDA-approved medicines: a focused review. J Basic Clin Physiol Pharmacol. 2023; 34(3):277-289. https://doi.org/10.1515/JBCPP-2022-0242/HTML

[155] Schöffmann G, Vettorato E, Burke JG, Gibson AJN, Clutton ER. The effects of age, isoflurane and sevoflurane on atracurium in lambs. Vet Anaesth Analg. 2012; 39(3):256-265. https://doi.org/10.1111/j.1467-2995.2011.00705.x

[156] Hunter JM, Blobner M. Under-dosing and over-dosing of neuromuscular blocking drugs and reversal agents: beware of the risks. Br J Anaesth. 2024; 132(3):461-465. https://doi.org/10.1016/j.bja.2023.12.001

[157] Zhang X, Chen S, Lu K, et al. Verapamil ameliorates motor neuron degeneration and improves lifespan in the SOD1G93A mouse model of ALS by enhancing autophagic flux. Aging Dis. 2019; 10(6):1159-1173. https://doi.org/10.14336/AD.2019.0228

[158] Liu Y, Liu X, Chen X, et al. Senolytic and senomorphic agent procyanidin C1 alleviates structural and functional decline in the aged retina. Proc Natl Acad Sci USA. 2024; 121(18) https://doi.org/10.1073/pnas.2311028121

[159] Shao M, Qiu Y, Shen M, et al. Procyanidin C1 inhibits bleomycin-induced pulmonary fibrosis in mice by selective clearance of senescent myofibroblasts. FASEB J. 2024; 38(13) https://doi.org/10.1096/fj.202302547RR

[160] Zhu WL, Zheng JY, Cai WW, et al. Correction: ligustilide improves aging-induced memory deficit by regulating mitochondrial related inflammation in SAMP8 mice. [Aging 12 (2019) (31753189)]. 2020; 12(6):5587. https://doi.org/10.18632/aging.102793 〈https://doi.org/10.18632/aging.102998〉(Aging).

[161] Zhu WL, Zheng JY, Cai WW, et al. Ligustilide improves aging-induced memory deficit by regulating mitochondrial related inflammation in SAMP8 mice. Aging. 2020; 12(4):3175-3189. https://doi.org/10.18632/aging.102793

[162] Dhas Y, Biswas N, MR D, Jones L, Biomedicine SA-M. Repurposing metabolic regulators: antidiabetic drugs as anticancer agents. & 2024, undefined. Springer. 2024; 5(1) https://doi.org/10.1186/s43556-024-00204-z

[163] Kulkarni A., Gubbi S., metabolism N. B.-C., & 2020, undefined. (2020). Benefits of metformin in attenuating the hallmarks of aging. Cell.Com, 32, 15-30. https://doi.org/10.1016/j.cmet.2020.04.001.

[164] Padki M., Aging I.S.-E. of G. and P., & 2022, undefined. (n.d.). Targeting Aging with Metformin (TAME). Springer. Retrieved January 8, 2025, from 〈https://link.springer.com/content/pdf/10.1007/978-3-030-22009-9_400.pdf〉.

[165] Baghdadi M, Nespital T, Monzó C, Deelen J, Grönke S, Partridge L. Intermittent rapamycin feeding recapitulates some effects of continuous treatment while maintaining lifespan extension. Mol Metab. 2024; 81:101902. https://doi.org/10.1016/J.MOLMET.2024.101902

[166] Chrienova Z, Nepovimova E, Kuca K. The role of mTOR in age-related diseases. J Enzym Inhib Med Chem. 2021; 36(1):1679-1693. https://doi.org/10.1080/14756366.2021.1955873

[167] Naeini SH, Mavaddatiyan L, Kalkhoran ZR, Taherkhani S, Talkhabi M. Alpha-ketoglutarate as a potent regulator for lifespan and healthspan: evidences and perspectives. Exp Gerontol. 2023; 175. https://doi.org/10.1016/j.exger.2023.112154

[168] Asadi Shahmirzadi A, Edgar D, Liao CY, et al. Alpha-Ketoglutarate, an endogenous metabolite, extends lifespan and compresses morbidity in aging mice. Cell Metab. 2020; 32(3):447-456.e6. https://doi.org/10.1016/J.CMET.2020.08.004

[169] Luo J., Zhang X., Li W., Lan Y., Li F., Xiao J., … Y. C.-P., & 2024, undefined. (n.d.). Pentagalloyl glucose enhanced the stress resistance to delay aging process in Caenorhabditis elegans. Elsevier. Retrieved January 7, 2025, from 〈https://www.sciencedirect.com/science/article/pii/S1359511324003179〉.

[170] Izadi M, Sadri N, Abdi A, et al. Longevity and anti-aging effects of curcumin supplementation. GeroScience. 2024; 46(3):2933-2950. https://doi.org/10.1007/s11357-024-01092-5

[171] Seong KM, Yu M, Lee KS, Park S, Jin YW, Min KJ. Curcumin mitigates accelerated aging after irradiation in drosophila by reducing oxidative stress. BioMed Res Int. 2015; 2015:425380. https://doi.org/10.1155/2015/425380

[172] Mahoney SA, Venkatasubramanian R, Darrah MA, et al. Intermittent supplementation with fisetin improves arterial function in old mice by decreasing cellular senescence. Aging Cell. 2024; 23(3) https://doi.org/10.1111/acel.14060

[173] Zhao R, Kou H, Jiang D, Wang F. Exploring the anti-aging effects of fisetin in telomerase-deficient progeria mouse model. PeerJ. 2023; 11:e16463. https://doi.org/10.7717/PEERJ.16463/SUPP-5

[174] Cui Z, Zhao X, Amevor FK, et al. Therapeutic application of quercetin in aging-related diseases: SIRT 1 as a potential mechanism. Front Immunol. 2022;13. https://doi.org/10.3389/fimmu.2022.943321

[175] Yang Y, Li Y, Du X, et al. Anti-Aging effects of quercetin in cladocera simocephalus vetulus using proteomics. ACS Omega. 2023; 8(20):17609-17619. https://doi.org/10.1021/acsomega.2c08242

[176] Liu W, Lin H, Mao Z, et al. Verapamil extends lifespan in caenorhabditis elegans by inhibiting calcineurin activity and promoting autophagy. Aging (Albany NY). 2020; 12(6):5300. https://doi.org/10.18632/AGING.102951

[177] Ni YQ, Liu YS. New insights into the roles and mechanisms of spermidine in aging and Age-Related diseases. Aging Dis. 2021; 12(8):1948-1963. https://doi.org/10.14336/AD.2021.0603

[178] Xu TT, Li H, Dai Z, et al. Spermidine and spermine delay brain aging by inducing autophagy in SAMP8 mice. Aging (Albany NY). 2020; 12(7):6401. https://doi.org/10.18632/AGING.103035

[179] Qi Z, Ji H, Le M, et al. Sulforaphane promotes C. Elegans longevity and healthspan via DAF-16/DAF-2 insulin/IGF-1 signaling. Aging. 2021; 13(2):1649-1670. https://doi.org/10.18632/AGING.202512

[180] Wang Z., Ma J., Sun Y., Jin Z., Zheng R., Li Y., … H. Y.-I., & 2024, undefined. (n.d.). Isorhapontigenin delays senescence and matrix degradation of nucleus pulposus cells via PI3K/AKT/mTOR-mediated autophagy pathway in vitro and alleviates. ElsevierZ Wang, J Ma, Y Sun, Z Jin, R Zheng, Y Li, H Yu, H Ye, Y Wu, X Ge, Z ChenInternational Immunopharmacology, 2024•Elsevier. Retrieved January 8, 2025, from 〈https://www.sciencedirect.com/science/article/pii/S1567576924012384〉.

[181] McIntyre RL, Denis SW, Kamble R, et al. Inhibition of the neuromuscular acetylcholine receptor with atracurium activates FOXO/DAF-16-induced longevity. Aging Cell. 2021; 20(8):e13381. https://doi.org/10.1111/ACEL.13381

[182] Zhang X, Zhu C, Zhang L, et al. RNA m6A methylation regulatory mechanism of resveratrol in premature senescence cells. Food Sci Nutr. 2024; 12(11):9238. https://doi.org/10.1002/FSN3.4487

[183] Xu X, Tao N, Sun C, et al. Ligustilide prevents thymic immune senescence by regulating thymosin β15-dependent spatial distribution of thymic epithelial cells. Phytomedicine Int J Phytother Phytopharm. 2024; 123. https://doi.org/10.1016/J.PHYMED.2023.155216

[184] Bhandari A, Sharma AK, Singh H, Singh A, Kaur G. Aging-related changes in metabolic indicators in female rats and their management with tinospora cordifolia. Biogerontology. 2022; 23(3):363-380. https://doi.org/10.1007/s10522-022-09962-1

[185] Xu Q, Fu Q, Li Z, et al. The flavonoid procyanidin C1 has senotherapeutic activity and increases lifespan in mice. Nat Metab. 2021; 3(12):1706. https://doi.org/10.1038/S42255-021-00491-8

[186] Peng L., Dan J., Huang W., Sang L., Tian H., … Z. L.-J. of, & 2024, undefined. (n.d.). The dual effects of Congea chinensis Moldenke on inhibiting tumor cell proliferation and delaying aging by activating TERT transcriptional activity. ElsevierL Peng, J Dan, W Huang, L Sang, H Tian, Z Li, W Li, J Liu, Y LuoJournal of Ethnopharmacology, 2024•Elsevier. Retrieved January 8, 2025, from 〈https://www.sciencedirect.com/science/article/pii/S0378874123009716〉.

[187] Liu C., Hua L., Biology Z.X.-R., & 2024, undefined. (n.d.). Synergistic impact of 25-hydroxyvitamin D concentrations and physical activity on delaying aging. ElsevierC Liu, L Hua, Z XinRedox Biolo gy, 2024•Elsevier. Retrieved January 8, 2025, from 〈https://www.sciencedirect.com/science/article/pii/S2213231724001666〉.

[188] Lu G, Wu Z, Shang J, Xie Z, Chen C, Zhang C. The effects of metformin on autophagy. Biomed Pharmacother. 2021; 137:111286. https://doi.org/10.1016/j.biopha.2021.111286

[189] Harrison DE, Strong R, Sharp ZD, et al. Rapamycin fed late in life extends lifespan in genetically heterogeneous mice. Nature. 2009; 460(7253):392-395. https://doi.org/10.1038/nature08221

[190] Slack C, Foley A, Partridge L. Activation of AMPK by the putative dietary restriction mimetic metformin is insufficient to extend lifespan in drosophila. PLoS ONE. 2012; 7(10):e47699. https://doi.org/10.1371/journal.pone.0047699

[191] Baskin KK, Winders BR, Olson EN. Muscle as a “Mediator” of systemic metabolism. Cell Metab. 2015; 21(2):237-248. https://doi.org/10.1016/j.cmet.2014.12.021

[192] Kang MJ, Moon JW, Lee JO, et al. Metformin induces muscle atrophy by transcriptional regulation of myostatin via HDAC6 and FoxO3a. J Cachex Sarcopenia Muscle. 2022; 13(1):605-620. https://doi.org/10.1002/jcsm.12833

[193] Suzuta S, Nishida H, Ozaki M, Kohno N, Le TD, Inoue YH. Metformin suppresses progression of muscle aging via activation of the AMP kinase-mediated pathways in drosophila adults. Eur Rev Med Pharmacol Sci. 2022; 26(21):8039-8056. https://doi.org/10.26355/eurrev_202211_30158

[194] Oka S, Hirai J, Yasukawa T, Nakahara Y, Inoue YH. A correlation of reactive oxygen species accumulation by depletion of superoxide dismutases with age-dependent impairment in the nervous system and muscles of drosophila adults. Biogerontology. 2015; 16(4):485-501. https://doi.org/10.1007/s10522-015-9570-3

[195] Le TD, Inoue YH. Sesamin activates Nrf2/Cnc-Dependent transcription in the absence of oxidative stress in drosophila adult brains. Antioxidants. 2021; 10(6):924. https://doi.org/10.3390/antiox10060924

[196] Bharath L., Agrawal M., McCambridge G., metabolism D. N.-C., & 2020, undefined. (2020). Metformin enhances autophagy and normalizes mitochondrial function to alleviate aging-associated inflammation. Cell.ComLP Bharath, M Agrawal, G McCambridge, DA Nicholas, H Hasturk, J Liu, K Jiang, R LiuCell Metabolism, 2020•cell.Com, 32, 44-55.e6. https://doi.org/10.1016/j.cmet.2020.04.015.

[197] Bu F, Yang M, Guo X, Huang W, Chen L. Multiple functions of ATG 8 family proteins in plant autophagy. Front Cell Dev Biol. 2020;8. https://doi.org/10.3389/fcell.2020.00466

[198] Lin L-Y, Chow H-X, Chen C-H, Mitsuda N, Chou W-C, Liu T-Y. Role of autophagy-related proteins ATG8f and ATG8h in the maintenance of autophagic activity in arabidopsis roots under phosphate starvation. Front Plant Sci. 2023; 14. https://doi.org/10.3389/fpls.2023.1018984

[199] Gao C, Fang L, Zhang H, Zhang W-S, Li X-O, Du S-Y. Metformin induces autophagy via the AMPK-mTOR signaling pathway in human hepatocellular carcinoma cells. Cancer Manag Res. 2020; 12:5803-5811. https://doi.org/10.2147/CMAR.S257966

[200] Amorim IS, Lach G, Gkogkas CG. The role of the eukaryotic translation initiation factor 4E (eIF4E) in neuropsychiatric disorders. Front Genet. 2018; 9. https://doi.org/10.3389/fgene.2018.00561

[201] Webb AE, Brunet A. FOXO transcription factors: key regulators of cellular quality control. Trends Biochem Sci. 2014; 39(4):159-169. https://doi.org/10.1016/j.tibs.2014.02.003

[202] Murtaza G, Khan AK, Rashid R, Muneer S, Hasan SMF, Chen J. FOXO transcriptional factors and Long-Term living. Oxid Med Cell Longev. 2017; 2017(1) https://doi.org/10.1155/2017/3494289

[203] Demontis F, Piccirillo R, Goldberg AL, Perrimon N. Mechanisms of skeletal muscle aging: insights from drosophila and mammalian models. Dis Models Mech. 2013. https://doi.org/10.1242/dmm.012559

[204] Zamanian MY, Golmohammadi M, Yumashev A, et al. Effects of metformin on cancers in experimental and clinical studies: focusing on autophagy and AMPK/mTOR signaling pathways. Cell Biochem Funct. 2024; 42(4) https://doi.org/10.1002/CBF.4071

[205] Petakh P, Kamyshna I, Kamyshnyi A. Gene expression of protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1), solute carrier family 2 member 1 (SLC2A1) and mechanistic target of rapamycin (MTOR) in metformin-treated type 2 diabetes patients with COVID-19: impact on inflammation markers. Inflammopharmacology. 2024; 32(1):885-891. https://doi.org/10.1007/S10787-023-01341-7

[206] Tee A. The target of rapamycin and mechanisms of cell growth. Int J Mol Sci. 2018; 19(3):880. https://doi.org/10.3390/ijms19030880

[207] Saxton RA, Sabatini DM. mTOR signaling in growth, metabolism, and disease. Cell. 2017; 168(6):960-976. https://doi.org/10.1016/j.cell.2017.02.004

[208] Valovka T, Verdier F, Cramer R, et al. Protein kinase c phosphorylates ribosomal protein S6 kinase βII and regulates its subcellular localization. Mol Cell Biol. 2003; 23(3):852-863. https://doi.org/10.1128/MCB.23.3.852-863.2003

[209] Jung C., Kim E., Song J., J. P.-E. & M., & 2019, undefined. (n.d.). Mitochondrial superoxide dismutase 2 mediates γ-irradiation-induced cancer cell invasion. Nature. Com. https://doi.org/10.1038/s12276-019-0207-5.

[210] Zhang Y, Liu Y, Liu X, et al. Exercise and metformin intervention prevents Lipotoxicity-Induced hepatocyte apoptosis by alleviating oxidative and ER stress and activating the AMPK/Nrf2/HO-1. wiley online library. 2022;2022. https://doi.org/10.1155/2022/2297268

[211] Ling NXY, Kaczmarek A, Hoque A, et al. mTORC1 directly inhibits AMPK to promote cell proliferation under nutrient stress. Nat Metab. 2020; 2(1):41-49. https://doi.org/10.1038/s42255-019-0157-1

[212] Caligaris M, Nicastro R, Hu Z, et al. Snf1/AMPK fine-tunes TORC1 signaling in response to glucose starvation. ELife. 2023; 12. https://doi.org/10.7554/eLife.84319

[213] Petrocelli JJ, McKenzie AI, de Hart NMMP, et al. Disuse-induced muscle fibrosis, cellular senescence, and senescence-associated secretory phenotype in older adults are alleviated during re-ambulation with metformin pre-treatment. Aging Cell. 2023; 22(11) https://doi.org/10.1111/ACEL.13936

[214] Witham MD, McDonald C, Wilson N, et al. Metformin and physical performance in older people with probable sarcopenia and physical prefrailty or frailty in england (MET-PREVENT): a double-blind, randomised, placebo-controlled trial. Lancet Healthy Longev. 2025; 6(3):100695. https://doi.org/10.1016/J.LANHL.2025.100695

[215] McKenzie AI, Mahmassani ZS, Petrocelli JJ, et al. Short-term exposure to a clinical dose of metformin increases skeletal muscle mitochondrial H2O2 emission and production in healthy, older adults: a randomized controlled trial. Exp Gerontol. 2022; 163:111804. https://doi.org/10.1016/J.EXGER.2022.111804

[216] Walton RG, Dungan CM, Long DE, et al. Metformin blunts muscle hypertrophy in response to progressive resistance exercise training in older adults: a randomized, double-blind, placebo-controlled, multicenter trial: the MASTERS trial. Aging Cell. 2019; 18(6) https://doi.org/10.1111/ACEL.13039

[217] Rosell-Díaz M, Petit-Gay A, Molas-Prat C, et al. Metformin-induced changes in the gut microbiome and plasma metabolome are associated with cognition in men. Metab Clin Exp. 2024; 157. https://doi.org/10.1016/J.METABOL.2024.155941

[218] Corley MJ, Pang APS, Shikuma CM, Ndhlovu LC. Cell-type specific impact of metformin on monocyte epigenetic age reversal in virally suppressed older people living with HIV. Aging Cell. 2024; 23(1):e13926. https://doi.org/10.1111/ACEL.13926;SUBPAGE:STRING:FULL

[219] Barzilai N, Crandall JP, Kritchevsky SB, Espeland MA. Metformin as a tool to target aging. Cell Metab. 2016; 23(6):1060. https://doi.org/10.1016/J.CMET.2016.05.011

[220] Kumari S, Bubak MT, Schoenberg HM, et al. Antecedent metabolic health and metformin (ANTHEM) aging study: rationale and study design for a randomized controlled trial. J Gerontol Ser A Biol Sci Med Sci. 2022; 77(12):2373-2377. https://doi.org/10.1093/GERONA/GLAB358

[221] Heckman-Stoddard BM, Crandall JP, Edelstein SL, et al. Randomized study of metformin and intensive lifestyle intervention on cancer incidence over 21 years of Follow-up in the diabetes prevention program. Cancer Prev Res. 2025; 18(7) 〈https://doi.org/10.1158/1940-6207.CAPR-23-0461/761281/AM/RANDOMIZED-STUDY-OF-METFORMIN-AND-INTENSIVE〉OF1-OF11.

[222] Martin-Montalvo A, Mercken EM, Mitchell SJ, et al. Metformin improves healthspan and lifespan in mice. Nat Commun. 2013; 4(1):2192. https://doi.org/10.1038/ncomms3192

[223] Cabreiro F, Au C, Leung K-Y, et al. Metformin retards aging in C. Elegans by altering microbial folate and methionine metabolism. Cell. 2013; 153(1):228-239. https://doi.org/10.1016/j.cell.2013.02.035

[224] Liu Y., Wang X., Jin C., Qiao J., Wang C., Jiang L., Phytomedicine S.Y.-, & 2024, undefined. (n.d.). Total ginsenosides extend healthspan of aging Drosophila by suppressing imbalances in intestinal stem cells and microbiota. Elsevier. Retrieved January 8, 2025, from 〈https://www.sciencedirect.com/science/article/pii/S094471132400309X〉.

[225] Golubev D, Platonova E, Zemskaya N, et al. Berberis vulgaris L. Extract supplementation exerts regulatory effects on the lifespan and healthspan of drosophila through its antioxidant activity depending on the sex. Biogerontology. 2024; 25(3):507-528. https://doi.org/10.1007/S10522-023-10083-6

[226] Zhu Y, Engmann M, Medina D, et al. Metformin treatment of juvenile mice alters aging-related developmental and metabolic phenotypes in sex-dependent and sex-independent manners. GeroScience. 2024; 46(3):3197-3218. https://doi.org/10.1007/s11357-024-01067-6

[227] Barzilai N, Crandall JP, Kritchevsky SB, Espeland MA. Metformin as a tool to target aging. Cell Metab. 2016; 23(6):1060-1065. https://doi.org/10.1016/j.cmet.2016.05.011

[228] Hardie DG, Ross FA, Hawley SA. AMPK: a nutrient and energy sensor that maintains energy homeostasis. Nat Rev Mol Cell Biol. 2012; 13(4):251-262. https://doi.org/10.1038/nrm3311

[229] Lu G, Wu Z, Shang J, Xie Z, Chen C, Zhang C. The effects of metformin on autophagy. Biomed Pharmacother. 2021; 137:111286. https://doi.org/10.1016/j.biopha.2021.111286

[230] Shang R, Miao J. Mechanisms and effects of metformin on skeletal muscle disorders. Front Neurol. 2023; 14:1275266. https://doi.org/10.3389/FNEUR.2023.1275266/XML