Network pharmacology-based identification of cardamomin as a ferroptosis inhibitor in ischemic stroke via SP1/ALOX5 axis

  • Pathol Res Pract. 2026 May 24:286:156558. doi: 10.1016/j.prp.2026.156558.
Libo Shao  1 ,  Xiaolin Gao  2 ,  Wenming Yu  1 ,  Wenwen Zhang  3 ,  Xing Lu  4
Affiliations
  • 1. Tuina Department, Jinan Hospital of Integrated Traditional Chinese and Western Medicine, Jinan City, Shandong 271100, China.
  • 2. The 7th Ward, Jinan Hospital of Integrated Traditional Chinese and Western Medicine, Jinan City, Shandong 271100, China.
  • 3. Tuina Department, Jinan Hospital of Integrated Traditional Chinese and Western Medicine, Jinan City, Shandong 271100, China. Electronic address: [email protected].
  • 4. Department of Acupuncture, Jinan Hospital of Integrated Traditional Chinese and Western Medicine, Jinan City, Shandong 271100, China.
PMID: 42208355 DOI: 10.1016/j.prp.2026.156558
Abstract

Background: Ischemic stroke (IS) causes high mortality and disability, yet current thrombolytic therapy is constrained by narrow time windows and side effects. Ferroptosis has recently as a critical pathological mechanism in cerebral ischemia-reperfusion injury, but effective modulators remain lacking. Cardamomin shows neuroprotective potential; however, its role in regulating Ferroptosis during IS is unknown and warrants urgent investigation.

Methods: Potential targets of cardamomin were identified using bioinformatics tools. KEGG enrichment analysis and molecular docking were performed. Cell viability and Apoptosis were assessed by MTT and flow cytometry. Inflammatory cytokines and Ferroptosis markers were detected by specific commercial kits. Western blot, dual-luciferase reporter assay, and chromatin immunoprecipitation were employed to investigate the SP1-ALOX5 regulatory axis. For in vivo validation, a middle cerebral artery occlusion model was established. Evaluations included neurological deficits, infarct volume by TTC staining, histopathology by HE staining, and molecular expression analysis.

Results: ALOX5 was identified as a key ferroptosis-related target. Cardamomin alleviated OGD/R-induced injury by restoring cell viability, suppressing Apoptosis and inflammation, and reversing ferroptotic changes. Molecular docking confirmed a strong binding affinity between cardamomin and ALOX5 (-7.6 kcal/mol). Mechanistically, transcription factor SP1 transcriptionally activated ALOX5 by binding to its promoter, and cardamomin downregulated the SP1/ALOX5 axis and subsequently activated the PI3K/Akt pathway. In vivo, cardamomin reduced cerebral infarction, improved neurological function, and inhibited inflammation and Ferroptosis.

Conclusion: Cardamomin attenuates neuronal Ferroptosis and ischemic injury by inhibiting the SP1/ALOX5 axis and activating the PI3K/Akt pathway, providing a novel therapeutic candidate for IS.

Keywords
Arachidonate 5-lipoxygenase; Cardamomin; Ferroptosis; Ischemic stroke; Specificity protein 1.
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