{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T11:18:05Z","timestamp":1783423085415,"version":"3.54.6"},"reference-count":29,"publisher":"IMR Press","issue":"6","license":[{"start":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T00:00:00Z","timestamp":1782345600000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["82274444"],"award-info":[{"award-number":["82274444"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Front. Biosci. (Landmark Ed)"],"abstract":"<jats:p>Background:Effective neuroprotective therapies for acute ischemic stroke (AIS) remain limited due to the complex interplay between neuroinflammation and apoptosis. Sinomenine (Sino), a bioactive alkaloid derived from Sinomenium acutum, exhibits anti-inflammatory and anti-apoptotic activities; however, its molecular targets and mechanisms in AIS remain unclear. This study aimed to identify potential targets and key pathways of Sino and validate its neuroprotective effects in AIS.Methods:A combined approach integrating network pharmacology, Mendelian randomization (MR), molecular docking, and in vivo validation was adopted. Potential targets of Sino and ischemic stroke were identified using public databases. Overlapping targets were analyzed through protein-protein interaction network construction and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Sprague-Dawley rats were randomly assigned to four groups, and a middle cerebral artery occlusion\/reperfusion (MCAO\/R) model was established (n = 12 per group): Sham, MCAO\/R, Sino (20 mg\/kg), and Sino +LY294002 (LY, 10 mg\/kg). Sino and Sino + LY were administered intraperitoneally within 6 h after surgery and once daily thereafter for three days. Sham and MCAO groups were given the same amount of physiological saline undergoing the same procedures. Sino was administered intraperitoneally within 6 h after surgery and once daily thereafter for three days. Neurological deficits, infarct volume, neuronal injury, apoptosis, activation of the phosphatidylinositol 3-kinase\/protein kinase B (PI3K\/Akt) pathway, and inflammatory responses were assessed using behavioral tests, 2,3,5-Triphenyltetrazolium chloride (TTC)\/Nissl\/Terminal deoxynucleotidyl transferase dUTP Nick End Labeling (TUNEL) staining, Western blotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA).Results:Twelve overlapping targets between Sino and ischemic stroke were identified, with Akt1 recognized as a central hub. Enrichment analysis highlighted the PI3K\/Akt pathway as a critical signaling axis, while MR analysis indicated a nominal association between Akt1 and ischemic stroke. Molecular docking predicted stable binding between Sino and Akt1. In MCAO\/R rats, Sino significantly improved neurological function, reduced infarct volumes, attenuated neuronal apoptosis, and increased neuronal survival. Mechanistically, Sino increased the p-PI3K\/PI3K and p-Akt\/Akt ratios, upregulated Bcl-2 expression, and decreased the expression of Bax, cleaved caspase-3, ionized calcium-binding adapter molecule 1 (Iba1), inducible nitric oxide synthase (iNOS), interleukin-1\u03b2 (IL-1\u03b2), interleukin-6 (IL-6), and tumor necrosis factor-\u03b1 (TNF-\u03b1). These beneficial effects were notably attenuated by LY.Conclusion:This study establishes PI3K\/Akt as a functionally necessary mediator of Sino\u2019s neuroprotection against cerebral ischemia\/reperfusion injury. The incomplete LY reversal indicates multi-target activity, supporting Sino\u2019s development as an adjunctive therapeutic candidate for ischemic stroke.<\/jats:p>","DOI":"10.31083\/fbl50007","type":"journal-article","created":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T10:50:51Z","timestamp":1783421451000},"source":"Crossref","is-referenced-by-count":0,"title":["Multi-Method Investigation of the Role of the PI3K\/Akt Pathway in Sinomenine-Mediated Neuroprotection Against Acute Ischemic Stroke"],"prefix":"10.31083","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-9049-0010","authenticated-orcid":true,"given":"Xinzuo","family":"Qin","sequence":"first","affiliation":[{"name":"Department of Neurology, Xuanwu Hospital, Capital Medical University, 100053 Beijing, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0737-9776","authenticated-orcid":true,"given":"Wancheng","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Neurology, Xuanwu Hospital, Capital Medical University, 100053 Beijing, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-8287-4915","authenticated-orcid":true,"given":"Xiaoqun","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Encephalopathy, Third Clinical Medical College of Beijing University of Chinese Medicine, 100029 Beijing, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5896-7869","authenticated-orcid":true,"given":"Yun","family":"Du","sequence":"additional","affiliation":[{"name":"Department of Neurology, Xuanwu Hospital, Capital Medical University, 100053 Beijing, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9225-5443","authenticated-orcid":true,"given":"Lulu","family":"Wen","sequence":"additional","affiliation":[{"name":"Department of Neurology, Xuanwu Hospital, Capital Medical University, 100053 Beijing, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6289-9654","authenticated-orcid":true,"given":"Miao","family":"Qu","sequence":"additional","affiliation":[{"name":"Department of Neurology, Xuanwu Hospital, Capital Medical University, 100053 Beijing, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"16242","published-online":{"date-parts":[[2026,6,25]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","unstructured":"Qin F, Zhao Y, Shang W, Zhao ZM, Qian X, Zhang BB, et al. Sinomenine relieves oxygen and glucose deprivation-induced microglial activation via inhibition of the SP1\/miRNA-183-5p\/I\u03baB-\u03b1 signaling pathway. Cellular and Molecular Biology (Noisy-le-Grand, France). 2018; 64: 140\u2013147.","DOI":"10.14715\/cmb\/2018.64.10.23"},{"key":"ref2","doi-asserted-by":"publisher","unstructured":"Campbell B, Khatri P. Stroke. Lancet (London, England). 2020; 396: 129\u2013142.","DOI":"10.1016\/S0140-6736(20)31179-X"},{"key":"ref3","doi-asserted-by":"publisher","unstructured":"Iadecola C, Anrather J. The immunology of stroke: from mechanisms to translation. Nature Medicine. 2011; 17: 796\u2013808.","DOI":"10.1038\/nm.2399"},{"key":"ref4","doi-asserted-by":"publisher","unstructured":"Zhu Y, Zhang J, Deng Q, Chen X. Mitophagy-associated programmed neuronal death and neuroinflammation. Frontiers in immunology. 2024; 15: 1460286.","DOI":"10.3389\/fimmu.2024.1460286"},{"key":"ref5","doi-asserted-by":"publisher","unstructured":"Fang Y, Gao S, Wang X, Cao Y, Lu J, Chen S, et al. Programmed Cell Deaths and Potential Crosstalk With Blood-Brain Barrier Dysfunction After Hemorrhagic Stroke. Frontiers in Cellular Neuroscience. 2020; 14: 68.","DOI":"10.3389\/fncel.2020.00068"},{"key":"ref6","doi-asserted-by":"publisher","unstructured":"Song G, Ouyang G, Bao S. The activation of Akt\/PKB signaling pathway and cell survival. Journal of Cellular and Molecular Medicine. 2005; 9: 59\u201371.","DOI":"10.1111\/j.1582-4934.2005.tb00337.x"},{"key":"ref7","doi-asserted-by":"publisher","unstructured":"Liu M, Huang X, Tian Y, Yan X, Wang F, Chen J, et al. Phosphorylated GSK-3\u03b2 protects stress-induced apoptosis of myoblasts via the PI3K\/Akt signaling pathway. Molecular Medicine Reports. 2020; 22: 317\u2013327.","DOI":"10.3892\/mmr.2020.11105"},{"key":"ref8","doi-asserted-by":"publisher","unstructured":"Xu H, Qin W, Hu X, Mu S, Zhu J, Lu W, et al. Lentivirus-mediated overexpression of OTULIN ameliorates microglia activation and neuroinflammation by depressing the activation of the NF-\u03baB signaling pathway in cerebral ischemia\/reperfusion rats. Journal of Neuroinflammation. 2018; 15: 83.","DOI":"10.1186\/s12974-018-1117-5"},{"key":"ref9","doi-asserted-by":"publisher","unstructured":"Guo N, Wang X, Xu M, Bai J, Yu H, Le Zhang. PI3K\/AKT signaling pathway: Molecular mechanisms and therapeutic potential in depression. Pharmacological Research. 2024; 206: 107300.","DOI":"10.1016\/j.phrs.2024.107300"},{"key":"ref10","doi-asserted-by":"publisher","unstructured":"Xu JL, Che HJ, Yang ZZ, Yang M. Research advances in the neuroprotective effect of natural drugs in ischemic stroke. Journal of QingDao University(Medical Sciences). 2025; 61: 312\u2013316.  (In Chinese)","DOI":"10.11712\/jms.2096-5532.2025.61.050"},{"key":"ref11","doi-asserted-by":"publisher","unstructured":"Yan M, Ni F, Xie X, Zhang C, Zhu J. The Role of Formononetin in Cerebral Ischemia-Reperfusion Injury: A New Mediator of c-Fos\/IL-10\/STAT3 Signaling Pathway. Frontiers in Bioscience (Landmark Edition). 2025; 30: 26274.","DOI":"10.31083\/fbl26274"},{"key":"ref12","doi-asserted-by":"publisher","unstructured":"Chen J, Zhu L, Chen Y, Liu Y, Chen W, Liu X, et al. Targeted neural stem cell-derived extracellular vesicles loaded with Sinomenine alleviate diabetic peripheral neuropathy via WNT5a\/TRPV1 pathway modulation. Journal of Nanobiotechnology. 2025; 23: 588.","DOI":"10.1186\/s12951-025-03678-3"},{"key":"ref13","doi-asserted-by":"publisher","unstructured":"Wang Q, Li XK. Immunosuppressive and anti-inflammatory activities of sinomenine. International Immunopharmacology. 2011; 11: 373\u2013376.","DOI":"10.1016\/j.intimp.2010.11.018"},{"key":"ref14","doi-asserted-by":"publisher","unstructured":"Hong H, Lu X, Lu Q, Huang C, Cui Z. Potential therapeutic effects and pharmacological evidence of sinomenine in central nervous system disorders. Frontiers in Pharmacology. 2022; 13: 1015035.","DOI":"10.3389\/fphar.2022.1015035"},{"key":"ref15","doi-asserted-by":"publisher","unstructured":"Qiu J, Yan Z, Tao K, Li Y, Li Y, Li J, et al. Sinomenine activates astrocytic dopamine D2 receptors and alleviates neuroinflammatory injury via the CRYAB\/STAT3 pathway after ischemic stroke in mice. Journal of Neuroinflammation. 2016; 13: 263.","DOI":"10.1186\/s12974-016-0739-8"},{"key":"ref16","doi-asserted-by":"publisher","unstructured":"Biose IJ, Chastain WH, Wang H, Ouvrier B, Bix GJ. Optimizing intraluminal monofilament model of ischemic stroke in middle-aged Sprague-Dawley rats. BMC Neuroscience. 2022; 23: 75.","DOI":"10.1186\/s12868-022-00764-2"},{"key":"ref17","doi-asserted-by":"publisher","unstructured":"Rogers DC, Campbell CA, Stretton JL, Mackay KB. Correlation between motor impairment and infarct volume after permanent and transient middle cerebral artery occlusion in the rat. Stroke. 1997; 28: 2060\u20132065; discussion 2066.","DOI":"10.1161\/01.str.28.10.2060"},{"key":"ref18","doi-asserted-by":"publisher","unstructured":"Longa EZ, Weinstein PR, Carlson S, Cummins R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke. 1989; 20: 84\u201391.","DOI":"10.1161\/01.str.20.1.84"},{"key":"ref19","doi-asserted-by":"publisher","unstructured":"Bederson JB, Pitts LH, Tsuji M, Nishimura MC, Davis RL, Bartkowski H. Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. Stroke. 1986; 17: 472\u2013476.","DOI":"10.1161\/01.str.17.3.472"},{"key":"ref20","doi-asserted-by":"publisher","unstructured":"Qin C, Yang S, Chu YH, Zhang H, Pang XW, Chen L, et al. Signaling pathways involved in ischemic stroke: molecular mechanisms and therapeutic interventions. Signal Transduction and Targeted Therapy. 2022; 7: 215.","DOI":"10.1038\/s41392-022-01064-1"},{"key":"ref21","doi-asserted-by":"publisher","unstructured":"Liu T, Li X, Zhou X, Chen W, Wen A, Liu M, et al. PI3K\/AKT signaling and neuroprotection in ischemic stroke: molecular mechanisms and therapeutic perspectives. Neural Regeneration Research. 2025; 20: 2758\u20132775.","DOI":"10.4103\/NRR.NRR-D-24-00568"},{"key":"ref22","doi-asserted-by":"publisher","unstructured":"Zhang P, He Y, Zhen Q, Zhang Y. Systematic Exploration of Potential Druggable Genes for Ischemic Stroke Employing Genome-Wide Mendelian Randomization Analysis. Brain and Behavior. 2025; 15: e70857.","DOI":"10.1002\/brb3.70857"},{"key":"ref23","doi-asserted-by":"publisher","unstructured":"Levinson S, Pulli B, Heit JJ. Neuroinflammation and acute ischemic stroke: impact on translational research and clinical care. Frontiers in Surgery. 2025; 12: 1501359.","DOI":"10.3389\/fsurg.2025.1501359"},{"key":"ref24","doi-asserted-by":"publisher","unstructured":"She R, Liu D, Liao J, Wang G, Ge J, Mei Z. Mitochondrial dysfunctions induce PANoptosis and ferroptosis in cerebral ischemia\/reperfusion injury: from pathology to therapeutic potential. Frontiers in Cellular Neuroscience. 2023; 17: 1191629.","DOI":"10.3389\/fncel.2023.1191629"},{"key":"ref25","doi-asserted-by":"publisher","unstructured":"Wang H, Li J, Zhang H, Wang M, Xiao L, Wang Y, et al. Regulation of microglia polarization after cerebral ischemia. Frontiers in Cellular Neuroscience. 2023; 17: 1182621.","DOI":"10.3389\/fncel.2023.1182621"},{"key":"ref26","unstructured":"Semba S, Itoh N, Ito M, Harada M, Yamakawa M. The in vitro and in vivo effects of 2-(4-morpholinyl)-8-phenyl-chromone (LY294002), a specific inhibitor of phosphatidylinositol 3'-kinase, in human colon cancer cells. Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. 2002; 8: 1957\u20131963."},{"key":"ref27","doi-asserted-by":"publisher","unstructured":"Walker EH, Pacold ME, Perisic O, Stephens L, Hawkins PT, Wymann MP, et al. Structural determinants of phosphoinositide 3-kinase inhibition by wortmannin, LY294002, quercetin, myricetin, and staurosporine. Molecular Cell. 2000; 6: 909\u2013919.","DOI":"10.1016\/s1097-2765(05)00089-4"},{"key":"ref28","doi-asserted-by":"publisher","unstructured":"O'Collins VE, Macleod MR, Donnan GA, Horky LL, Van Der Worp BH, Howells DW. 1,026 experimental treatments in acute stroke. Annals of Neurology. 2006; 59: 467\u2013477.","DOI":"10.1002\/ana.20741"},{"key":"ref29","doi-asserted-by":"publisher","unstructured":"Chamorro \u00c1, Dirnagl U, Urra X, Planas AM. Neuroprotection in acute stroke: targeting excitotoxicity, oxidative and nitrosative stress, and inflammation. The Lancet. Neurology. 2016; 15: 869\u2013881.","DOI":"10.1016\/S1474-4422(16)00114-9"}],"container-title":["Frontiers in Bioscience-Landmark"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.imrpress.com\/journal\/FBL\/31\/6\/10.31083\/FBL50007","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T10:52:26Z","timestamp":1783421546000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.imrpress.com\/journal\/FBL\/31\/6\/10.31083\/FBL50007"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,25]]},"references-count":29,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2026]]}},"alternative-id":["S2768-6701(26)02145-3"],"URL":"https:\/\/doi.org\/10.31083\/fbl50007","relation":{},"ISSN":["2768-6701","2768-6698"],"issn-type":[{"value":"2768-6701","type":"print"},{"value":"2768-6698","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,25]]},"article-number":"50007"}}