{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T20:28:55Z","timestamp":1792700935349,"version":"4.5.0"},"reference-count":40,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T00:00:00Z","timestamp":1773878400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100018537","name":"National Science and Technology Major Project","doi-asserted-by":"crossref","award":["2023ZD0505605"],"award-info":[{"award-number":["2023ZD0505605"]}],"id":[{"id":"10.13039\/501100018537","id-type":"DOI","asserted-by":"crossref"}]},{"name":"\u201cSharp Spear and Leading Goose\u201d R&D Program of Zhejiang Province","award":["2024C03105"],"award-info":[{"award-number":["2024C03105"]}]},{"name":"Traditional Chinese Medicine Administration of Zhejiang Province","award":["2025ZL032"],"award-info":[{"award-number":["2025ZL032"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>In the pathological cascade of cerebral ischemia, the pyroptosis axis mediated by the NLRP3 inflammasome in activated microglia is a core link driving neuroinflammation and secondary brain injury. Quercetin has been proven to possess multi-target neuroprotective activity, and its anti-inflammatory effect has attracted particular attention. However, direct molecular evidence is lacking regarding how quercetin precisely regulates the NLRP3\/Caspase-1\/GSDMD core pyroptosis axis in microglia in cerebral ischemia models and whether it can directly target NLRP3 to inhibit this axis, thereby alleviating cerebral ischemic injury. This study aimed to investigate the molecular mechanism by which quercetin alleviates cerebral ischemic injury through inhibiting the pyroptosis axis, combining cellular and animal models with molecular docking and molecular dynamics simulations. The oxygen-glucose deprivation (OGD) model of BV2 microglia and the photothrombotic (PT) model of focal cortical ischemia in male C57BL\/6 mice were used to detect the ameliorative effect of quercetin on cerebral ischemia-related injury through cellular and animal experiments. AutoDock Vina 1.5.7 and GROMACS 2025.3 software were employed for molecular docking and molecular dynamics simulations, respectively, to analyze the binding mode and complex stability between quercetin and the NLRP3 protein. The results showed that quercetin could significantly ameliorate OGD-induced injury in BV2 cells and downregulate the expression of pyroptosis and inflammation-related proteins and factors. Meanwhile, it relieved motor dysfunction in PT mice, attenuated cortical neuronal injury, and inhibited the activation of the cerebral pyroptosis axis. At the molecular level, molecular simulation predictions indicated that quercetin might specifically bind to the NACHT domain of the NLRP3 protein, forming a complex with a stable conformation, and van der Waals interactions served as the main driving force for binding. This study confirmed that quercetin can directly bind to the NLRP3 protein and alleviate cerebral ischemia-induced inflammatory injury by inhibiting the activation of the NLRP3\/Caspase-1\/GSDMD pyroptosis axis and the release of downstream inflammatory factors. Combined with the molecular simulation results, a predictive hypothesis is proposed: direct binding of quercetin to the NLRP3 protein is one of its core mechanisms of action. These findings provide direct experimental evidence for the development of NLRP3-based drugs against ischemic brain injury.<\/jats:p>","DOI":"10.3390\/cells15060552","type":"journal-article","created":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T15:38:01Z","timestamp":1773934681000},"page":"552","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Quercetin Alleviates Cerebral Ischemia-Induced Neuroinflammation by Inhibiting Microglia-Mediated NLRP3\/Caspase-1\/GSDMD Pathway"],"prefix":"10.3390","volume":"15","author":[{"given":"Da","family":"Shen","sequence":"first","affiliation":[{"name":"College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China"},{"name":"Key Laboratory of TCM Encephalopathy of Zhejiang Province, Zhejiang Chinese Medical University, Hangzhou 310053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weiao","family":"Kong","sequence":"additional","affiliation":[{"name":"College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China"},{"name":"Key Laboratory of TCM Encephalopathy of Zhejiang Province, Zhejiang Chinese Medical University, Hangzhou 310053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoke","family":"Qiu","sequence":"additional","affiliation":[{"name":"College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China"},{"name":"Key Laboratory of TCM Encephalopathy of Zhejiang Province, Zhejiang Chinese Medical University, Hangzhou 310053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huiling","family":"Yuan","sequence":"additional","affiliation":[{"name":"College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China"},{"name":"Key Laboratory of TCM Encephalopathy of Zhejiang Province, Zhejiang Chinese Medical University, Hangzhou 310053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanyi","family":"Wu","sequence":"additional","affiliation":[{"name":"College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China"},{"name":"Key Laboratory of TCM Encephalopathy of Zhejiang Province, Zhejiang Chinese Medical University, Hangzhou 310053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lefan","family":"Huang","sequence":"additional","affiliation":[{"name":"College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China"},{"name":"Key Laboratory of TCM Encephalopathy of Zhejiang Province, Zhejiang Chinese Medical University, Hangzhou 310053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zixin","family":"Yin","sequence":"additional","affiliation":[{"name":"College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China"},{"name":"Key Laboratory of TCM Encephalopathy of Zhejiang Province, Zhejiang Chinese Medical University, Hangzhou 310053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5149-5881","authenticated-orcid":false,"given":"Lisheng","family":"Chu","sequence":"additional","affiliation":[{"name":"Department of Physiology, Zhejiang Chinese Medical University, Hangzhou 310053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lijun","family":"Ge","sequence":"additional","affiliation":[{"name":"College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China"},{"name":"Key Laboratory of TCM Encephalopathy of Zhejiang Province, Zhejiang Chinese Medical University, Hangzhou 310053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"108","DOI":"10.3810\/hp.2013.04.1060","article-title":"Management of Acute Ischemic Stroke","volume":"41","author":"Vidal","year":"2013","journal-title":"Hosp. Pract."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1007\/s00401-018-1930-z","article-title":"Post-stroke inflammation-target or tool for therapy?","volume":"137","author":"Lambertsen","year":"2019","journal-title":"Acta Neuropathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1038\/s41583-020-00396-7","article-title":"Encouraging an excitable brain state: Mechanisms of brain repair in stroke","volume":"22","author":"Joy","year":"2021","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3033","DOI":"10.1161\/STROKEAHA.121.032241","article-title":"Cerebroprotection for Acute Ischemic Stroke: Looking Ahead","volume":"52","author":"Lyden","year":"2021","journal-title":"Stroke"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1007\/s13105-021-00817-w","article-title":"Pyroptosis in stroke-new insights into disease mechanisms and therapeutic strategies","volume":"77","author":"Gou","year":"2021","journal-title":"J. Physiol. Biochem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"660","DOI":"10.1038\/nature15514","article-title":"Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death","volume":"526","author":"Shi","year":"2015","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"113776","DOI":"10.1016\/j.jep.2021.113776","article-title":"Salvianolic Acids for Injection alleviates cerebral ischemia\/reperfusion injury by switching M1\/M2 phenotypes and inhibiting NLRP3 inflammasome\/pyroptosis axis in microglia in vivo and in vitro","volume":"270","author":"Ma","year":"2021","journal-title":"J. Ethnopharmacol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e790","DOI":"10.1038\/cddis.2013.326","article-title":"Intravenous immunoglobulin suppresses NLRP1 and NLRP3 inflammasome-mediated neuronal death in ischemic stroke","volume":"4","author":"Fann","year":"2013","journal-title":"Cell Death Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"101239","DOI":"10.1016\/j.redox.2019.101239","article-title":"Programmed necrotic cell death of macrophages: Focus on pyroptosis, necroptosis, and parthanatos","volume":"26","author":"Robinson","year":"2019","journal-title":"Redox Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.plaphy.2021.05.023","article-title":"The role of quercetin in plants","volume":"166","author":"Singh","year":"2021","journal-title":"Plant Physiol. Biochem."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Batiha, G.E., Beshbishy, A.M., Ikram, M., Mulla, Z.S., El-Hack, M.E.A., Taha, A.E., Algammal, A.M., and Elewa, Y.H.A. (2020). The Pharmacological Activity, Biochemical Properties, and Pharmacokinetics of the Major Natural Polyphenolic Flavonoid: Quercetin. Foods, 9.","DOI":"10.3390\/foods9030374"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"278","DOI":"10.1080\/10286020.2021.1949302","article-title":"Quercetin attenuates ischemia reperfusion injury by protecting the blood-brain barrier through Sirt1 in MCAO rats","volume":"24","author":"Yang","year":"2021","journal-title":"J. Asian Nat. Prod. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"108436","DOI":"10.1016\/j.jnutbio.2020.108436","article-title":"Quercetin protects against cerebral ischemia\/reperfusion and oxygen glucose deprivation\/reoxygenation neurotoxicity","volume":"83","author":"Wang","year":"2020","journal-title":"J. Nutr. Biochem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/s13062-024-00470-5","article-title":"Pharmacological inhibition of mTORC1 reduces neural death and damage volume after MCAO by modulating microglial reactivity","volume":"19","author":"Rubio","year":"2024","journal-title":"Biol. Direct"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"mjac077","DOI":"10.1093\/jmcb\/mjac077","article-title":"Argon mitigates post-stroke neuroinflammation by regulating M1\/M2 polarization and inhibiting NF-\u03baB\/NLRP3 inflammasome signaling","volume":"14","author":"Xue","year":"2023","journal-title":"J. Mol. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"115653","DOI":"10.1016\/j.biopha.2023.115653","article-title":"Quercetin improves cerebral ischemia\/reperfusion injury by promoting microglia\/macrophages M2 polarization via regulating PI3K\/Akt\/NF-\u03baB signaling pathway","volume":"168","author":"Li","year":"2023","journal-title":"Biomed. Pharmacother."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"115847","DOI":"10.1016\/j.biopha.2023.115847","article-title":"Flavonoids and ischemic stroke-induced neuroinflammation: Focus on the glial cells","volume":"170","author":"Lu","year":"2024","journal-title":"Biomed. Pharmacother."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1743","DOI":"10.1007\/s11030-023-10684-w","article-title":"Network pharmacology, molecular docking, and molecular dynamics simulation to elucidate the mechanism of anti-aging action of Tinospora cordifolia","volume":"28","author":"Bisht","year":"2024","journal-title":"Mol. Divers."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Liu, L., Jiao, Y., Yang, M., Wu, L., Long, G., and Hu, W. (2023). Network Pharmacology, Molecular Docking and Molecular Dynamics to Explore the Potential Immunomodulatory Mechanisms of Deer Antler. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241210370"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1002\/kjm2.12136","article-title":"miR-15a regulates oxygen glucose deprivation\/reperfusion (OGD\/R)-induced neuronal injury by targeting BDNF","volume":"36","author":"Hu","year":"2020","journal-title":"Kaohsiung J. Med. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"ENEURO.0244-22.2022","DOI":"10.1523\/ENEURO.0244-22.2022","article-title":"Photothrombotic Middle Cerebral Artery Occlusion in Mice: A Novel Model of Ischemic Stroke","volume":"10","author":"Conti","year":"2023","journal-title":"eNeuro"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"134110","DOI":"10.1063\/5.0018516","article-title":"Heterogeneous parallelization and acceleration of molecular dynamics simulations in GROMACS","volume":"153","author":"Zhmurov","year":"2020","journal-title":"J. Chem. Phys."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3696","DOI":"10.1021\/acs.jctc.5b00255","article-title":"ff14SB: Improving the Accuracy of Protein Side Chain and Backbone Parameters from ff99SB","volume":"11","author":"Maier","year":"2015","journal-title":"J. Chem. Theory Comput."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1002\/wcms.81","article-title":"The ORCA program system","volume":"2","author":"Neese","year":"2012","journal-title":"Wiley Interdiscip. Rev. Comput. Mol. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"082503","DOI":"10.1063\/5.0216272","article-title":"A comprehensive electron wavefunction analysis toolbox for chemists, Multiwfn","volume":"161","author":"Lu","year":"2024","journal-title":"J. Chem. Phys."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"935","DOI":"10.1038\/nrd1549","article-title":"Docking and scoring in virtual screening for drug discovery: Methods and applications","volume":"3","author":"Kitchen","year":"2004","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1258","DOI":"10.1021\/ci700044s","article-title":"Analysis of HIV wild-type and mutant structures via in silico docking against diverse ligand libraries","volume":"47","author":"Chang","year":"2007","journal-title":"J. Chem. Inf. Model."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1097\/00004647-200404000-00001","article-title":"Cerebral ischemia and reperfusion: The pathophysiologic concept as a basis for clinical therapy","volume":"24","author":"Schaller","year":"2004","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_29","first-page":"6","article-title":"Effects of quercetin on long-term learning and memory and PARP-1\/AIF signaling pathway in neonatal rats with hypoxic-ischemic brain injury","volume":"34","author":"Yang","year":"2016","journal-title":"J. Clin. Pediatr."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"116894","DOI":"10.1016\/j.biopha.2024.116894","article-title":"Artemisinin attenuated ischemic stroke induced pyroptosis by inhibiting ROS\/TXNIP\/NLRP3\/Caspase-1 signaling pathway","volume":"177","author":"Wang","year":"2024","journal-title":"Biomed. Pharmacother."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"113399","DOI":"10.1016\/j.expneurol.2020.113399","article-title":"Kv13 channel blockade alleviates cerebral ischemia\/reperfusion injury by reshaping M1\/M2 phenotypes and compromising the activation of NLRP3 inflammasome in microglia","volume":"332","author":"Ma","year":"2020","journal-title":"Exp. Neurol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5823823","DOI":"10.1155\/2018\/5823823","article-title":"Recent Advances in the Molecular Mechanisms Underlying Pyroptosis in Sepsis","volume":"2018","author":"Gao","year":"2018","journal-title":"Mediat. Inflamm."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1111\/imr.12534","article-title":"Molecular mechanisms and functions of pyroptosis, inflammatory caspases and inflammasomes in infectious diseases","volume":"277","author":"Man","year":"2017","journal-title":"Immunol. Rev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.neures.2020.07.011","article-title":"Intrathecal administration of the extracellular vesicles derived from human Wharton\u2019s jelly stem cells inhibit inflammation and attenuate the activity of inflammasome complexes after spinal cord injury in rats","volume":"170","author":"Noori","year":"2021","journal-title":"Neurosci. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1097\/MBC.0000000000001370","article-title":"Serum levels of inflammatory markers, sP-selectin, IL-1\u03b2, IL-6, and hsCRP are positively correlated with tissue factor transcript level of peripheral blood mononuclear cells in stroke","volume":"36","author":"Ebrahimi","year":"2025","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"112050","DOI":"10.1016\/j.jpsychores.2025.112050","article-title":"Association between serum inflammatory cytokines levels and post-stroke depression among stroke patients: A meta-analysis and systematic review","volume":"190","author":"Zhou","year":"2025","journal-title":"J. Psychosom. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1089\/jir.2024.0176","article-title":"Correlations of Levels of Peripheral Blood Inflammatory Cytokines and Serum Lipoprotein-Associated Phospholipase-A2 with Acute Ischemic Stroke and Their Early Prognostic Value","volume":"45","author":"Zhong","year":"2025","journal-title":"J. Interferon Cytokine Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2183026","DOI":"10.1155\/2016\/2183026","article-title":"ROS-Mediated NLRP3 Inflammasome Activation in Brain, Heart, Kidney, and Testis Ischemia\/Reperfusion Injury","volume":"2016","author":"Letteria","year":"2016","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.bbr.2017.06.027","article-title":"Nrf2 inhibits NLRP3 inflammasome activation through regulating Trx1\/TXNIP complex in cerebral ischemia reperfusion injury","volume":"336","author":"Hou","year":"2018","journal-title":"Behav. Brain Res. Int. J."},{"key":"ref_40","first-page":"3760","article-title":"Potential molecular mechanisms and clinical application feasibility of quercetin in the treatment of traumatic brain injury","volume":"23","author":"Huang","year":"2019","journal-title":"Chin. J. Tissue Eng. Res."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/6\/552\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,21]],"date-time":"2026-03-21T05:16:27Z","timestamp":1774070187000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/6\/552"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,19]]},"references-count":40,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["cells15060552"],"URL":"https:\/\/doi.org\/10.3390\/cells15060552","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,19]]}}}