{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T00:31:29Z","timestamp":1783557089699,"version":"3.55.0"},"reference-count":44,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,8,21]],"date-time":"2025-08-21T00:00:00Z","timestamp":1755734400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Ischemic stroke is a leading cause of mortality and disability worldwide, with limited therapeutic options and high rates of recurrence. Mitochondrial dysfunction plays a critical role in neuronal injury during ischemia-reperfusion, making mitochondrial autophagy a potential therapeutic target. Gypenoside XLIX, a major active metabolite of Gynostemma pentaphyllum, exhibits antioxidant and organ-protective properties, but its effects on neuronal mitochondrial damage in stroke remain unclear. This study aimed to explore the neuroprotective mechanisms of Gypenoside XLIX in ischemic stroke, focusing on the PI3K\/AKT\/FOXO1 signaling pathway.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Neuroprotective effects were evaluated in oxygen-glucose deprivation (OGD) neuronal cells and middle cerebral artery occlusion (MCAO) rat models. Cell viability, apoptosis, ROS production, mitochondrial membrane potential, and autophagic flux were assessed by CCK-8, flow cytometry, ELISA, TMRE staining, immunofluorescence, and Western blotting. Signaling pathway involvement was examined using PI3K inhibitor LY294002, AKT activator SC79, and FOXO1 knockdown.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Gypenoside XLIX significantly improved neuronal viability (p &amp;lt; 0.01), reduced apoptosis (p &amp;lt; 0.01), and decreased ROS levels (p &amp;lt; 0.001) in OGD cells. It enhanced p-PI3K and p-AKT expression while suppressing FOXO1 (p &amp;lt; 0.05), promoted Beclin-1, LC3, PINK1, and Parkin expression (p &amp;lt; 0.001), and reduced p62 (p &amp;lt; 0 .01). In MCAO rats, Gypenoside XLIX decreased infarct volume (p &amp;lt; 0.001), brain edema (p &amp;lt; 0.01), and TUNEL-positive cells (p &amp;lt; 0.001), while elevating mitochondrial membrane potential and antioxidant enzyme levels (SOD, GSH-Px, CAT; all p &amp;lt; 0.001).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Gypenoside XLIX alleviates ischemic stroke injury by activating the PI3K\/AKT\/FOXO1 pathway, enhancing mitochondrial autophagy, and reducing oxidative stress, supporting its potential as a novel neuroprotective agent in stroke management.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fphar.2025.1600435","type":"journal-article","created":{"date-parts":[[2025,8,21]],"date-time":"2025-08-21T05:28:12Z","timestamp":1755754092000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["Gypenoside XLIX inhibiting PI3K\/AKT\/FOXO1 signaling pathway mediated neuronal mitochondrial autophagy to improve patients with ischemic stroke"],"prefix":"10.3389","volume":"16","author":[{"given":"Yonglei","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongdie","family":"Mao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengguang","family":"Sha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jishuai","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Cai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rong","family":"Xi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenzhu","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoling","family":"Yin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Changyun","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,8,21]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"107848","DOI":"10.1016\/j.pharmthera.2021.107848","article-title":"Targeting autophagy in ischemic stroke: from molecular mechanisms to clinical therapeutics","volume":"225","author":"Ajoolabady","year":"2021","journal-title":"Pharmacol. Ther."},{"key":"B2","doi-asserted-by":"publisher","first-page":"168","DOI":"10.1186\/s12974-020-01853-x","article-title":"TREM2 activation attenuates neuroinflammation and neuronal apoptosis via PI3K\/Akt pathway after intracerebral hemorrhage in mice","volume":"17","author":"Chen","year":"2020","journal-title":"J. Neuroinflammation"},{"key":"B3","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1038\/s41419-022-04963-x","article-title":"The role of the PI3K\/AKT signalling pathway in the corneal epithelium: recent updates","volume":"13","author":"Chen","year":"2022","journal-title":"Cell Death Dis."},{"key":"B4","doi-asserted-by":"publisher","first-page":"662","DOI":"10.1016\/j.jgr.2023.04.007","article-title":"Discovery and validation of PURA as a transcription target of 20(S)-protopanaxadiol: implications for the treatment of cognitive dysfunction","volume":"47","author":"Chen","year":"","journal-title":"J. Ginseng Res."},{"key":"B5","doi-asserted-by":"publisher","first-page":"e3325","DOI":"10.1002\/brb3.3325","article-title":"Gandouling alleviates nerve injury through PI3K\/Akt\/FoxO1 and Sirt1\/FoxO1 signaling pathway to inhibit autophagy in the rats model of wilson's disease","volume":"13","author":"Chen","year":"","journal-title":"Brain Behav."},{"key":"B6","doi-asserted-by":"publisher","first-page":"183","DOI":"10.56434\/j.arch.esp.urol.20247702.24","article-title":"Effect of miR-1297 on kidney injury in rats with diabetic nephropathy through the PTEN\/PI3K\/AKT pathway","volume":"77","author":"Chen","year":"2024","journal-title":"Arch. Esp. Urol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"4433","DOI":"10.1080\/14786419.2020.1723093","article-title":"Dammarane-type saponins from Gynostemma pentaphyllum and their cytotoxicities","volume":"35","author":"Cui","year":"2021","journal-title":"Nat. Prod. Res."},{"key":"B8","doi-asserted-by":"publisher","first-page":"4670","DOI":"10.3390\/s22134670","article-title":"Stroke risk prediction with machine learning techniques","volume":"22","author":"Dritsas","year":"2022","journal-title":"Sensors (Basel)"},{"key":"B9","first-page":"500","article-title":"Network pharmacology analysis and in vitro validation of the active ingredients and potential mechanisms of gynostemma pentaphyllum against esophageal cancer","volume-title":"Comb. Chem. High Throughput Screen","author":"Guo","year":"2024"},{"key":"B10","doi-asserted-by":"publisher","first-page":"5165631","DOI":"10.1155\/2020\/5165631","article-title":"Identification and analysis of components in yizhi granule and cynomolgus monkey plasma after oral administration by UPLC\/ESI-Q-TOF MS and their protective effects on PC12 cells","volume":"2020","author":"Hao","year":"2020","journal-title":"J. Anal. Methods Chem."},{"key":"B11","doi-asserted-by":"publisher","first-page":"1250","DOI":"10.1093\/jpp\/rgab058","article-title":"Ganoderma lucidum ameliorates the diabetic nephropathy via down-regulatory effect on TGF\u03b2-1 and TLR-4\/NF\u03baB signalling pathways","volume":"73","author":"Hassan","year":"2021","journal-title":"J. Pharm. Pharmacol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"1654","DOI":"10.1097\/CCM.0000000000004597","article-title":"Management of acute ischemic stroke","volume":"48","author":"Herpich","year":"2020","journal-title":"Crit. Care Med."},{"key":"B13","doi-asserted-by":"publisher","first-page":"818","DOI":"10.1111\/bcpt.14007","article-title":"Optimizing myocardial cell protection with xanthine derivative KMUP-3 potentiates autophagy through the PI3K\/Akt\/eNOS axis","volume":"134","author":"Huang","year":"2024","journal-title":"Basic Clin. Pharmacol. Toxicol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"118721","DOI":"10.1016\/j.lfs.2020.118721","article-title":"Gypenoside inhibits ox-LDL uptake and foam cell formation through enhancing Sirt1-FOXO1 mediated autophagy flux restoration","volume":"264","author":"Hui","year":"2021","journal-title":"Life Sci."},{"key":"B15","doi-asserted-by":"publisher","first-page":"6285134","DOI":"10.1155\/2018\/6285134","article-title":"Isolation, structures, and bioactivities of the polysaccharides from Gynostemma pentaphyllum (Thunb.) makino: a review","volume":"2018","author":"Ji","year":"2018","journal-title":"Biomed. Res. Int."},{"key":"B16","doi-asserted-by":"publisher","first-page":"444","DOI":"10.1038\/s41419-022-04906-6","article-title":"Mitochondrial autophagy: molecular mechanisms and implications for cardiovascular disease","volume":"13","author":"Li","year":"","journal-title":"Cell Death Dis."},{"key":"B17","doi-asserted-by":"publisher","first-page":"711","DOI":"10.3390\/cells11040711","article-title":"The role of mitophagy in viral infection","volume":"11","author":"Li","year":"","journal-title":"Cells"},{"key":"B18","doi-asserted-by":"publisher","first-page":"953","DOI":"10.1142\/S0192415X23500441","article-title":"The role of Gynostemma pentaphyllum in regulating hyperlipidemia","volume":"51","author":"Li","year":"2023","journal-title":"Am. J. Chin. Med."},{"key":"B19","doi-asserted-by":"publisher","first-page":"1485","DOI":"10.14670\/HH-18-737","article-title":"Urolithin A attenuates bupivacaine-induced neurotoxicity in SH-SY5Y cells by regulating the SIRT1-activated PI3K\/AKT pathway","volume":"39","author":"Liu","year":"2024","journal-title":"Histol. Histopathol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"174501","DOI":"10.1016\/j.ejphar.2021.174501","article-title":"Gypenoside XLIX loaded nanoparticles targeting therapy for renal fibrosis and its mechanism","volume":"910","author":"Liu","year":"2021","journal-title":"Eur. J. Pharmacol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"7963","DOI":"10.3390\/molecules27227963","article-title":"A new therapeutic trend: natural medicine for ameliorating ischemic stroke via PI3K\/Akt signaling pathway","volume":"27","author":"Liu","year":"2022","journal-title":"Molecules"},{"key":"B22","first-page":"834","article-title":"Effects of gypenoside XLIX on fatty liver cell gene expression in vitro: a genome-wide analysis","volume":"15","author":"Liu","year":"","journal-title":"Am. J. Transl. Res."},{"key":"B23","doi-asserted-by":"publisher","first-page":"5509","DOI":"10.1002\/ptr.7994","article-title":"Advances of phytotherapy in ischemic stroke targeting PI3K\/Akt signaling","volume":"37","author":"Liu","year":"","journal-title":"Phytother. Res."},{"key":"B24","doi-asserted-by":"publisher","first-page":"599","DOI":"10.1016\/S1875-5364(23)60395-4","article-title":"Dammarane-type triterpenoid saponins isolated from Gynostemma pentaphyllum ameliorate liver fibrosis via agonizing PP2C\u03b1 and inhibiting deposition of extracellular matrix","volume":"21","author":"Liu","year":"","journal-title":"Chin. J. Nat. Med."},{"key":"B25","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1038\/s41419-021-03723-7","article-title":"Ablation of dynamin-related protein 1 promotes diabetes-induced synaptic injury in the hippocampus","volume":"12","author":"Park","year":"2021","journal-title":"Cell Death Dis."},{"key":"B26","doi-asserted-by":"publisher","first-page":"2347","DOI":"10.1007\/s12015-021-10212-z","article-title":"Oxidative stress enhances autophagy-mediated death of stem cells through Erk1\/2 signaling pathway - implications for neurotransplantations","volume":"17","author":"Prakash","year":"2021","journal-title":"Stem Cell Rev. Rep."},{"key":"B27","doi-asserted-by":"publisher","first-page":"386","DOI":"10.1212\/CON.0000000000000833","article-title":"Ischemic stroke in young adults","volume":"26","author":"Putaala","year":"2020","journal-title":"Contin. (Minneap Minn)"},{"key":"B28","doi-asserted-by":"publisher","first-page":"268","DOI":"10.1212\/CON.0000000000000840","article-title":"Update on treatment of acute ischemic stroke","volume":"26","author":"Rabinstein","year":"2020","journal-title":"Contin. (Minneap Minn)"},{"key":"B29","doi-asserted-by":"publisher","first-page":"134","DOI":"10.1007\/s00018-023-04777-w","article-title":"Atrial natriuretic peptide stimulates autophagy\/mitophagy and improves mitochondrial function in chronic heart failure","volume":"80","author":"Raffa","year":"2023","journal-title":"Cell Mol. Life Sci."},{"key":"B30","doi-asserted-by":"publisher","first-page":"138","DOI":"10.1016\/j.redox.2018.07.004","article-title":"Melatonin protects mouse granulosa cells against oxidative damage by inhibiting FOXO1-mediated autophagy: implication of an antioxidation-independent mechanism","volume":"18","author":"Shen","year":"2018","journal-title":"Redox Biol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"e106283","DOI":"10.15252\/embj.2020106283","article-title":"ATAD3B is a mitophagy receptor mediating clearance of oxidative stress-induced damaged mitochondrial DNA","volume":"40","author":"Shu","year":"2021","journal-title":"EMBO J."},{"key":"B32","doi-asserted-by":"publisher","first-page":"6249","DOI":"10.3390\/molecules26206249","article-title":"Progress in the medicinal value, bioactive compounds, and pharmacological activities of Gynostemma pentaphyllum","volume":"26","author":"Su","year":"2021","journal-title":"Molecules"},{"key":"B33","doi-asserted-by":"publisher","first-page":"e231455","DOI":"10.1001\/jamanetworkopen.2023.1455","article-title":"Estimated burden of stroke in China in 2020","volume":"6","author":"Tu","year":"2023","journal-title":"JAMA Netw. Open"},{"key":"B34","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1186\/s12868-022-00740-w","article-title":"Enhanced autophagy interacting proteins negatively correlated with the activation of apoptosis-related caspase family proteins after focal ischemic stroke of young rats","volume":"23","author":"Wang","year":"2022","journal-title":"BMC Neurosci."},{"key":"B35","doi-asserted-by":"publisher","first-page":"137439","DOI":"10.1016\/j.neulet.2023.137439","article-title":"Neuroprotective effect of 20 (S) - protopanaxadiol (PPD) attenuates NLRP3 inflammasome-mediated microglial pyroptosis in vascular dementia rats","volume":"814","author":"Wang","year":"2023","journal-title":"Neurosci. Lett."},{"key":"B36","doi-asserted-by":"publisher","first-page":"1243353","DOI":"10.3389\/fphar.2024.1243353","article-title":"Gypenoside induces apoptosis by inhibiting the PI3K\/AKT\/mTOR pathway and enhances T-cell antitumor immunity by inhibiting PD-L1 in gastric cancer","volume":"15","author":"Wu","year":"2024","journal-title":"Front. Pharmacol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"118066","DOI":"10.1016\/j.jep.2024.118066","article-title":"Hypolipidemic effect and gut microbiota regulation of gypenoside aglycones in rats fed a high-fat diet","volume":"328","author":"Xie","year":"","journal-title":"J. Ethnopharmacol."},{"key":"B38","doi-asserted-by":"publisher","first-page":"117186","DOI":"10.1016\/j.jep.2023.117186","article-title":"Saponins derived from Gynostemma pentaphyllum regulate triglyceride and cholesterol metabolism and the mechanisms: a review","volume":"319","author":"Xie","year":"","journal-title":"J. Ethnopharmacol."},{"key":"B39","doi-asserted-by":"publisher","first-page":"111420","DOI":"10.1016\/j.intimp.2023.111420","article-title":"Gypenoside XLIX attenuates sepsis-induced splenic injury through inhibiting inflammation and oxidative stress","volume":"127","author":"Xu","year":"2024","journal-title":"Int. Immunopharmacol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"3239","DOI":"10.1111\/cns.14256","article-title":"LIPUS-SCs-Exo promotes peripheral nerve regeneration in cavernous nerve crush injury-induced ED rats via PI3K\/Akt\/FoxO signaling pathway","volume":"29","author":"Ye","year":"2023","journal-title":"CNS Neurosci. Ther."},{"key":"B41","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1186\/s12906-020-2865-1","article-title":"Germacrone protects against oxygen-glucose deprivation\/reperfusion injury by inhibiting autophagy processes in PC12 cells","volume":"20","author":"Zhang","year":"2020","journal-title":"BMC Complement. Med. Ther."},{"key":"B42","doi-asserted-by":"publisher","first-page":"1891","DOI":"10.1083\/jcb.201804101","article-title":"Somatic autophagy of axonal mitochondria in ischemic neurons","volume":"218","author":"Zheng","year":"2019","journal-title":"J. Cell Biol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"111872","DOI":"10.1016\/j.intimp.2024.111872","article-title":"Gypenoside XLIX alleviates acute liver injury: emphasis on NF-\u03baB\/PPAR-\u03b1\/NLRP3 pathways","volume":"131","author":"Zhou","year":"2024","journal-title":"Int. Immunopharmacol."},{"key":"B44","doi-asserted-by":"publisher","first-page":"221","DOI":"10.3892\/ol.2024.14353","article-title":"MicroRNA-373-3p inhibits the proliferation and invasion of non-small-cell lung cancer cells by targeting the GAB2\/PI3K\/AKT pathway","volume":"27","author":"Zhu","year":"2024","journal-title":"Oncol. Lett."}],"container-title":["Frontiers in Pharmacology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2025.1600435\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,8,21]],"date-time":"2025-08-21T05:28:13Z","timestamp":1755754093000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2025.1600435\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,21]]},"references-count":44,"alternative-id":["10.3389\/fphar.2025.1600435"],"URL":"https:\/\/doi.org\/10.3389\/fphar.2025.1600435","relation":{},"ISSN":["1663-9812"],"issn-type":[{"value":"1663-9812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,21]]},"article-number":"1600435"}}