{ "abstract": "Background: Ischemic stroke, a leading cause of global mortality and disability, results from cerebral blood flow disruption, driving metabolic and neuronal dysfunction. Current therapies-thrombolysis, thrombectomy, and neuroprotectants-are constrained by narrow time windows and hemorrhagic risks. Notoginsenoside R1 (NGR1), a bioactive saponin from Panax notoginseng, exhibits multimodal neuroprotection, yet its mechanistic interplay with metabolic pathways remains unclear.\n\nObjective: We investigated NGR1’s neuroprotective efficacy in ischemic stroke, focusing on its modulation of the CKLF1/CCR5 axis and metabolic reprogramming.\n\nMethods: This study employs a multi-tiered approach integrating in vitro experiments, in vivo animal research, computationally validated target simulations, and gene knockout techniques to systematically elucidate NGR1’s regulatory role in key indicators during ischemic stroke treatment. Molecular docking and kinetic modelling first predicted NGR1-CKLF1 binding, subsequently validated via Cellular Thermal Shift Assay (CETSA) and Drug Affinity Responsive Target Stability (DARTS) experiments. In vitro experiments employed a PC12 cell oxygen-glucose deprivation (OGD) model to evaluate NGR1’s neuroprotective effects. CCK-8 assays, live-dead Cell staining, metabolic assay kits, TUNEL and JC-1 staining, Western blot (WB) analysis, and immunofluorescence (IF) co-localization revealed in vitro metabolic reprogramming mechanisms. In vivo experiments employed a permanent middle cerebral artery occlusion (pMCAO) model to mimic human ischemic pathophysiology. Neuronal metabolic reprogramming mechanisms following NGR1 treatment were elucidated via TTC staining, neurobehavioral testing, Nissl staining, Enzyme-linked immunosorbent assay (ELISA) assays, immunohistochemical (IHC), WB, IF, and Co-immunoprecipitation (Co-IP). Pharmacodynamic evaluation in the CKLF1−/− rat middle cerebral artery occlusion model, employing ELISA assays and WB analysis, confirmed NGR1’s specificity towards CKLF1.\n\nResults: Docking and MD predicted a stable NGR1–CKLF1 interaction; CETSA/DARTS supported a direct physical interaction, but binding-affinity constants were not determined. In vitro , NGR1 mitigated OGD-induced metabolic dysregulation, inhibited apoptosis, and enhanced neuronal survival. In vivo experiments demonstrated that NGR1 reduced infarct volume, improved neurological behavior scores, mitigated Nissl body damage, and alleviated inflammatory responses. Mechanistically, NGR1 inhibits downstream AMPK/mTOR/HIF-1α-driven neuronal metabolic reprogramming characterized by glycolytic activation by disrupting CKLF1-CCR5 binding. CKLF1 knockout abolished NGR1’s therapeutic effects and its role in neuronal metabolic reprogramming, confirming CKLF1 dependency.\n\nConclusion: NGR1 alleviates ischemic neuronal injury, at least in part, by interfering with CKLF1/CCR5-associated signaling, thereby attenuating pathological metabolic reprogramming, neuroinflammation, and apoptosis. Clinical relevance requires further validation.", "abstract_shown": true, "affiliations": [ "School of Pharmacy, Hunan Engineering Technology Center of Standardization and Function of Chinese Herbal Decoction Pieces, Hunan University of Chinese Medicine", "School of Chinese Materia Medica, Guangzhou University of Chinese Medicine", "Department of Pharmacy, Changsha Hospital for Maternal & Child Healthcare Affiliated to Hunan Normal University", "Department of Pharmacy, The First Hospital of Lanzhou University", "Key Laboratory of Cosmetic, China National Light Industry, Beijing Technology and Business University", "Nephrology Department, Xiangtan Central Hospital", "State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica & Neuroscience Center, Chinese Academy of Medical Sciences and Peking Union Medical College", "National Key Laboratory Cultivation Base of Chinese Medicinal Powder & Innovative Medicinal Jointly Established By Province and Ministry, Technology Innovation Center, Hunan University of Chinese Medicine" ], "article_type": "research article", "arxiv": null, "author_total": 16, "authors": [ { "affs": [ 0, 1 ], "name": "Hanlong Wang" }, { "affs": [ 2 ], "name": "Shasha Liu" }, { "affs": [ 3 ], "name": "Chen Chen" }, { "affs": [ 0 ], "name": "Junpeng Long" }, { "affs": [ 0, 1 ], "name": "Qian Yan" }, { "affs": [ 0 ], "name": "Yuting Lin" }, { "affs": [ 0 ], "name": "Shuqin Li" }, { "affs": [ 0 ], "name": "Yanting Huang" }, { "affs": [ 0 ], "name": "Songwei Yang" }, { "affs": [ 0 ], "name": "Meiyu Lin" }, { "affs": [ 0 ], "name": "Xuan Liu" }, { "affs": [ 4 ], "name": "Fan Yi" }, { "affs": [ 5 ], "name": "Yong Tan" }, { "affs": [ 0, 1, 6 ], "name": "Naihong Chen" }, { "affs": [ 0, 7 ], "name": "Qidi Ai" }, { "affs": [ 0 ], "name": "Yantao Yang" } ], "bibtex": "https://research.draconitic.com/paper/61sr3pct62gnx.bib", "cited_by": 0, "doi": "10.3389/fphar.2026.1964218", "fulltext_url": "https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13724343/?tool=EBI", "id": "61sr3pct62gnx", "issns": [ "1663-9812" ], "landing_url": "https://www.frontiersin.org/articles/10.3389/fphar.2026.1964218/full", "language": "eng", "licence": "CC-BY-4.0", "licence_label": "CC BY 4.0", "licence_source": "article", "licence_url": "https://creativecommons.org/licenses/by/4.0/", "metadata_licence": null, "native_subjects": [ [ "heading", "Original Research" ], [ "openalex", "1312|Ginseng Biological Effects and Applications" ] ], "openalex": "W7222915990", "pdf_url": "https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2026.1964218/pdf", "pmcid": "PMC13724343", "pmid": "42973581", "published": "2026-10-19", "publisher": "Frontiers Media SA", "ris": "https://research.draconitic.com/paper/61sr3pct62gnx.ris", "sources": [ { "first_seen": "2026-10-30T12:20:06Z", "native_id": "10.3389/fphar.2026.1964218", "source": "crossref", "url": "https://www.frontiersin.org/articles/10.3389/fphar.2026.1964218/full" }, { "first_seen": "2026-11-03T15:20:51Z", "native_id": "PMC:PMC13724343", "source": "europepmc", "url": "https://europepmc.org/article/PMC/PMC13724343" }, { "first_seen": "2026-11-04T12:22:56Z", "native_id": "PMC13724343", "source": "pmc", "url": "https://doi.org/10.3389/fphar.2026.1964218" }, { "first_seen": "2026-11-04T15:20:48Z", "native_id": "MED:42973581", "source": "europepmc", "url": "https://europepmc.org/article/MED/42973581" } ], "subjects": [ { "field": "biochemistry", "origin": "openalex:1312|Ginseng Biological Effects and Applications", "subfield": "molecular-biology" } ], "title": "Notoginsenoside R1 attenuates ischemic stroke via CKLF1-dependent suppression of neuronal metabolic reprogramming", "title_text": "Notoginsenoside R1 attenuates ischemic stroke via CKLF1-dependent suppression of neuronal metabolic reprogramming", "type": "peer-reviewed", "updated_at": "2026-11-06T12:20:51Z", "url": "https://research.draconitic.com/paper/61sr3pct62gnx", "venue": "Frontiers in Pharmacology", "versions": [] }