{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T17:32:45Z","timestamp":1786383165862,"version":"build-2736575974"},"reference-count":42,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T00:00:00Z","timestamp":1786320000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"\n Objectives<\/jats:title>\n To investigate whether selected Malaysian plant extracts modulate the antioxidant-autophagy axis to enhance neuronal and skeletal muscle protection.<\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n Four standardized extracts (\n Persicaria minor, Eurycoma longifolia<\/jats:italic>\n (EL)\n , Labisia pumila,<\/jats:italic>\n and\n Ipomoea aquatica<\/jats:italic>\n ) were evaluated for antioxidant activity at 100, 200, and 300 \u03bcg\/mL using ABTS, DPPH, and ORAC assays. The most active extract was further examined in SH-SY5Y neuronal-like cells and differentiated C2C12 myotubes. Cell viability was assessed using CCK-8, oxidative DNA damage was measured by 8-OHdG ELISA, and autophagy-related changes were evaluated by LC3\/p62 immunofluorescence and western blot analysis of LC3-II and p62 with chloroquine (CQ) as a lysosomal inhibitor.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Among the four extracts, EL exhibited the strongest antioxidant activity, achieving the highest values in ABTS (107.59\u2009\u00b1\u20097.98 \u03bcmol Trolox\/g), DPPH (27.28\u2009\u00b1\u20091.29 \u03bcmol Trolox\/g), and ORAC (81.12\u2009\u00b1\u20094.81 \u03bcmol Trolox\/g) at 300 \u03bcg\/mL, also showing the lowest IC\u2085\u2080 (167.22, 97.64, and 67.90 \u03bcg\/mL for ABTS, DPPH, and ORAC, respectively), with regression analyses confirming significant dose-response relationships (R\u00b2\u2009>\u20090.96,\n p<\/jats:italic>\n \u2009<\u20090.05). CCK-8 analysis showed that EL at 100\u2013300 \u03bcg\/mL did not cause overt cytotoxicity in either cell model after 24 h treatment (\n p<\/jats:italic>\n \u2009>\u20090.2), whereas the positive cytotoxicity control markedly reduced viability (\n p<\/jats:italic>\n \u2009<\u20090.001). 8-OHdG ELISA showed that EL reduced oxidative DNA damage, with significant reductions at 200 and 300 \u03bcg\/mL in both SH-SY5Y and C2C12 cells (\n p<\/jats:italic>\n \u2009<\u20090.05). H\n 2<\/jats:sub>\n O\n 2<\/jats:sub>\n markedly increased 8-OHdG levels in both models (\n p<\/jats:italic>\n \u2009<\u20090.001). Immunofluorescence analysis showed concentration-dependent modulation of LC3 and p62 autophagy-related markers. In SH-SY5Y cells, treatment with EL induced a dose-dependent increase in LC3 puncta (\n p<\/jats:italic>\n \u2009<\u20090.001) and a significant reduction in p62 fluorescence (\n p<\/jats:italic>\n \u2009<\u20090.01). Similar trends were observed in C2C12 myotubes, where LC3 puncta formation was enhanced (\n p<\/jats:italic>\n \u2009<\u20090.05), and p62 levels decreased (\n p<\/jats:italic>\n \u2009<\u20090.01). Western blot validation further showed that EL increased LC3-II abundance and that EL\u2009+\u2009CQ further increased LC3-II accumulation compared with CQ alone, particularly at 300 \u03bcg\/mL (\n p<\/jats:italic>\n \u2009<\u20090.05). p62 decreased under basal EL treatment but showed a non-canonical decrease in the CQ-only group and was therefore interpreted cautiously.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n EL showed the strongest antioxidant activity among the tested Malaysian plant extracts and modulated autophagy-related cellular markers without overt cytotoxicity. These findings identify EL as a promising candidate for further investigation in oxidative stress- and proteostasis-related cellular protection.<\/jats:p>\n <\/jats:sec>","DOI":"10.1371\/journal.pone.0355828","type":"journal-article","created":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T17:25:26Z","timestamp":1786382726000},"page":"e0355828","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["A double-duty champion: Eurycoma longifolia counters oxidative stress and unlocks autophagy to safeguard neurons and muscle cells"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-7481-0428","authenticated-orcid":true,"given":"Qing","family":"Huang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shinong","family":"Gu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hengzhi","family":"Deng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annie","family":"George","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8117-6440","authenticated-orcid":true,"given":"Ashril","family":"Yusof","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,8,10]]},"reference":[{"key":"pone.0355828.ref001","first-page":"1","article-title":"Biologically active compounds of malaysian medicinal and aromatic plants.","volume":"127","author":"N Manshoor","year":"2025","journal-title":"Prog Chem Org Nat Prod"},{"issue":"10","key":"pone.0355828.ref002","doi-asserted-by":"crossref","first-page":"2499","DOI":"10.1007\/s00204-023-03562-9","article-title":"Reactive oxygen species, toxicity, oxidative stress, and antioxidants: chronic diseases and aging","volume":"97","author":"K Jomova","year":"2023","journal-title":"Arch Toxicol"},{"key":"pone.0355828.ref003","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.ejmech.2015.04.040","article-title":"The role of antioxidants in the chemistry of oxidative stress: a review","volume":"97","author":"AM Pisoschi","year":"2015","journal-title":"Eur J Med Chem"},{"issue":"3","key":"pone.0355828.ref004","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1007\/s00204-020-02689-3","article-title":"Antioxidants and antioxidant methods: an updated overview","volume":"94","author":"\u0130 Gulcin","year":"2020","journal-title":"Arch Toxicol"},{"issue":"1","key":"pone.0355828.ref005","doi-asserted-by":"crossref","first-page":"16","DOI":"10.3390\/nu16010016","article-title":"Efficacy of probiotic supplements on brain-derived neurotrophic factor, inflammatory biomarkers, oxidative stress and cognitive function in patients with Alzheimer\u2019s dementia: a 12-week randomized, double-blind active-controlled study","volume":"16","author":"Y-C Hsu","year":"2023","journal-title":"Nutrients"},{"key":"pone.0355828.ref006","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1042\/bse0550119","article-title":"Autophagy and ageing: implications for age-related neurodegenerative diseases","volume":"55","author":"B Carroll","year":"2013","journal-title":"Essays Biochem"},{"issue":"4","key":"pone.0355828.ref007","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1007\/s13105-024-01039-6","article-title":"Role of ROS and autophagy in the pathological process of atherosclerosis","volume":"80","author":"L Zhu","year":"2024","journal-title":"J Physiol Biochem"},{"key":"pone.0355828.ref008","doi-asserted-by":"crossref","first-page":"2450748","DOI":"10.1155\/2018\/2450748","article-title":"The role of free radicals in autophagy regulation: implications for ageing","volume":"2018","author":"M Pajares","year":"2018","journal-title":"Oxid Med Cell Longev"},{"key":"pone.0355828.ref009","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.jprot.2013.01.004","article-title":"Protein oxidation in aging and the removal of oxidized proteins","volume":"92","author":"A H\u00f6hn","year":"2013","journal-title":"J Proteomics"},{"issue":"1","key":"pone.0355828.ref010","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1080\/15548627.2018.1511504","article-title":"TFEB-driven endocytosis coordinates MTORC1 signaling and autophagy","volume":"15","author":"IC Nnah","year":"2019","journal-title":"Autophagy"},{"issue":"2","key":"pone.0355828.ref011","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1038\/ncb2152","article-title":"AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1","volume":"13","author":"J Kim","year":"2011","journal-title":"Nat Cell Biol"},{"key":"pone.0355828.ref012","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1146\/annurev-pharmtox-011112-140320","article-title":"Role of nrf2 in oxidative stress and toxicity","volume":"53","author":"Q Ma","year":"2013","journal-title":"Annu Rev Pharmacol Toxicol"},{"issue":"1","key":"pone.0355828.ref013","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1186\/s12906-023-04290-4","article-title":"Ethanol extract of Andrographis paniculata alleviates aluminum-induced neurotoxicity and cognitive impairment through regulating the p62-keap1-Nrf2 pathway","volume":"23","author":"J Ma","year":"2023","journal-title":"BMC Complement Med Ther"},{"issue":"7","key":"pone.0355828.ref014","doi-asserted-by":"crossref","first-page":"1221","DOI":"10.1080\/15548627.2019.1659617","article-title":"m6A mRNA methylation controls autophagy and adipogenesis by targeting Atg5 and Atg7","volume":"16","author":"X Wang","year":"2020","journal-title":"Autophagy"},{"key":"pone.0355828.ref015","doi-asserted-by":"crossref","first-page":"109659","DOI":"10.1016\/j.cbi.2021.109659","article-title":"Fluoride regulates chondrocyte proliferation and autophagy via PI3K\/AKT\/mTOR signaling pathway","volume":"349","author":"L Ma","year":"2021","journal-title":"Chem Biol Interact"},{"key":"pone.0355828.ref016","first-page":"8603602","article-title":"A review of Malaysian medicinal plants with potential anti-inflammatory activity","volume":"2018","author":"FI Abu Bakar","year":"2018","journal-title":"Adv Pharmacol Sci"},{"key":"pone.0355828.ref017","doi-asserted-by":"crossref","first-page":"643119","DOI":"10.3389\/fphar.2021.643119","article-title":"Immunomodulatory effects and mechanisms of curcuma species and their bioactive compounds: a review","volume":"12","author":"JI Yuandani","year":"2021","journal-title":"Front Pharmacol"},{"issue":"27","key":"pone.0355828.ref018","doi-asserted-by":"crossref","first-page":"9805","DOI":"10.1080\/10408398.2023.2217264","article-title":"Nutritional and medicinal plants as potential sources of enzyme inhibitors toward the bioactive functional foods: an updated review","volume":"64","author":"H Saleem","year":"2024","journal-title":"Crit Rev Food Sci Nutr"},{"key":"pone.0355828.ref019","doi-asserted-by":"crossref","unstructured":"Varghese CP, Murugaiyah V, Parasuraman S, Christina AJM. Bioactive phytoconstituents and biological activities of Polygonum minus Huds. In: Bioactive compounds in the storage organs of plants; 2024. 1001 p.","DOI":"10.1007\/978-3-031-44746-4_48"},{"issue":"10","key":"pone.0355828.ref020","doi-asserted-by":"crossref","first-page":"512","DOI":"10.3923\/pjbs.2024.512.517","article-title":"Estimation of antioxidant, anticancer and cytotoxic properties of Eurycoma longifolia jack","volume":"27","author":"T Taechowisan","year":"2024","journal-title":"Pak J Biol Sci"},{"issue":"6","key":"pone.0355828.ref021","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1515\/jbcpp-2017-0160","article-title":"Phenolic-rich extracts of Eurycoma longifolia and Cylicodiscus gabunensis inhibit enzymes responsible for the development of erectile dysfunction and are antioxidants","volume":"29","author":"G Oboh","year":"2018","journal-title":"J Basic Clin Physiol Pharmacol"},{"issue":"4","key":"pone.0355828.ref022","first-page":"682","article-title":"Medicinal phytochemical and pharmacological properties of kesum (Polygonum minus Linn.): a mini review","volume":"6","author":"G Narasimhulu","year":"2014","journal-title":"Int J Pharm Pharm Sci"},{"key":"pone.0355828.ref023","doi-asserted-by":"crossref","first-page":"114423","DOI":"10.1016\/j.bbr.2023.114423","article-title":"Persicaria minor ameliorates the cognitive function of chronic cerebral hypoperfusion rats: metabolomic analysis and potential mechanisms","volume":"447","author":"LW Chun","year":"2023","journal-title":"Behav Brain Res"},{"issue":"2","key":"pone.0355828.ref024","doi-asserted-by":"crossref","first-page":"275","DOI":"10.31083\/j.rcm.2020.02.50","article-title":"Antioxidative and anti-inflammatory activities of Polygonum minus: a review of literature","volume":"21","author":"AA Hamid","year":"2020","journal-title":"Rev Cardiovasc Med"},{"key":"pone.0355828.ref025","doi-asserted-by":"crossref","first-page":"9928199","DOI":"10.1155\/2021\/9928199","article-title":"A review on therapeutic effects of Labisia pumila on female reproductive diseases","volume":"2021","author":"AA Zakaria","year":"2021","journal-title":"Biomed Res Int"},{"issue":"10","key":"pone.0355828.ref026","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0139831","article-title":"Water spinach, Ipomoea aquatic (Convolvulaceae), ameliorates lead toxicity by inhibiting oxidative stress and apoptosis","volume":"10","author":"S Dewanjee","year":"2015","journal-title":"PLoS One"},{"key":"pone.0355828.ref027","doi-asserted-by":"crossref","first-page":"1304903","DOI":"10.3389\/fnut.2023.1304903","article-title":"Chemical and biochemical characterization of Ipomoea aquatica: genoprotective potential and inhibitory mechanism of its phytochemicals against \u03b1-amylase and \u03b1-glucosidase","volume":"10","author":"K Saikia","year":"2023","journal-title":"Front Nutr"},{"issue":"6","key":"pone.0355828.ref028","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0325505","article-title":"Autophagy across tissues of aging mice","volume":"20","author":"JM Carosi","year":"2025","journal-title":"PLoS One"},{"key":"pone.0355828.ref029","doi-asserted-by":"crossref","first-page":"109895","DOI":"10.1016\/j.biopha.2020.109895","article-title":"Scoparone improves hepatic inflammation and autophagy in mice with nonalcoholic steatohepatitis by regulating the ROS\/P38\/Nrf2 axis and PI3K\/AKT\/mTOR pathway in macrophages","volume":"125","author":"B Liu","year":"2020","journal-title":"Biomed Pharmacother"},{"issue":"1","key":"pone.0355828.ref030","doi-asserted-by":"crossref","first-page":"11818","DOI":"10.1038\/s41598-018-30365-1","article-title":"Exercise prevents impaired autophagy and proteostasis in a model of neurogenic myopathy","volume":"8","author":"JC Campos","year":"2018","journal-title":"Sci Rep"},{"issue":"19","key":"pone.0355828.ref031","article-title":"Nobiletin prevents D-galactose-induced C2C12 cell aging by improving mitochondrial function","volume":"23","author":"HH Wang","year":"2022","journal-title":"Int J Mol Sci"},{"issue":"11","key":"pone.0355828.ref032","doi-asserted-by":"crossref","first-page":"7355","DOI":"10.1007\/s12035-019-1605-3","article-title":"Cholinergic differentiation of human neuroblastoma SH-SY5Y cell line and its potential use as an in vitro model for Alzheimer\u2019s disease studies","volume":"56","author":"LM de Medeiros","year":"2019","journal-title":"Mol Neurobiol"},{"issue":"15","key":"pone.0355828.ref033","doi-asserted-by":"crossref","first-page":"8713","DOI":"10.3390\/ijms23158713","article-title":"Effect of physical activity\/exercise on oxidative stress and inflammation in muscle and vascular aging","volume":"23","author":"M El Assar","year":"2022","journal-title":"Int J Mol Sci"},{"issue":"1","key":"pone.0355828.ref034","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1007\/s11033-024-09639-7","article-title":"Curcumin protects against aging-related stress and dysfunction through autophagy activation in rat brain","volume":"51","author":"A Singh","year":"2024","journal-title":"Mol Biol Rep"},{"issue":"7","key":"pone.0355828.ref035","doi-asserted-by":"crossref","first-page":"3380","DOI":"10.3390\/ijms22073380","article-title":"Analytical methods used in determining antioxidant activity: a review","volume":"22","author":"IG Munteanu","year":"2021","journal-title":"Int J Mol Sci"},{"issue":"5","key":"pone.0355828.ref036","doi-asserted-by":"crossref","first-page":"1768","DOI":"10.1021\/jf803011r","article-title":"Comparative study of antioxidant properties and total phenolic content of 30 plant extracts of industrial interest using DPPH, ABTS, FRAP, SOD, and ORAC assays","volume":"57","author":"S Dudonn\u00e9","year":"2009","journal-title":"J Agric Food Chem"},{"issue":"24","key":"pone.0355828.ref037","doi-asserted-by":"crossref","first-page":"7275","DOI":"10.1113\/JP275072","article-title":"The role of declining adaptive homeostasis in ageing","volume":"595","author":"LCD Pomatto","year":"2017","journal-title":"J Physiol"},{"issue":"4","key":"pone.0355828.ref038","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1207\/s15328015tlm1804_1","article-title":"ANCOVA, selection bias, statistical equating, and effect size: recommendations for publication","volume":"18","author":"JA Colliver","year":"2006","journal-title":"Teach Learn Med"},{"issue":"10","key":"pone.0355828.ref039","doi-asserted-by":"crossref","first-page":"1342","DOI":"10.1093\/gerona\/glw242","article-title":"Lysosomal proteolysis is associated with exercise-induced improvement of mitochondrial quality control in aged hippocampus","volume":"72","author":"L Luo","year":"2017","journal-title":"J Gerontol A Biol Sci Med Sci"},{"key":"pone.0355828.ref040","doi-asserted-by":"crossref","first-page":"176823","DOI":"10.1016\/j.ejphar.2024.176823","article-title":"Spermidine as an epigenetic regulator of autophagy in neurodegenerative disorders","volume":"979","author":"S Satarker","year":"2024","journal-title":"Eur J Pharmacol"},{"issue":"5","key":"pone.0355828.ref041","doi-asserted-by":"crossref","first-page":"2619","DOI":"10.3390\/ijms23052619","article-title":"Exercise and training regulation of autophagy markers in human and rat skeletal muscle","volume":"23","author":"J Botella","year":"2022","journal-title":"Int J Mol Sci"},{"issue":"11","key":"pone.0355828.ref042","doi-asserted-by":"crossref","first-page":"3813","DOI":"10.1080\/15548627.2021.1872187","article-title":"Crocetin promotes clearance of amyloid-\u03b2 by inducing autophagy via the STK11\/LKB1-mediated AMPK pathway","volume":"17","author":"A Wani","year":"2021","journal-title":"Autophagy"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0355828","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T17:25:33Z","timestamp":1786382733000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0355828"}},"subtitle":[],"editor":[{"given":"Irina V.","family":"Lebedeva","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,8,10]]},"references-count":42,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8,10]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0355828","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,10]]}}}