Abstract: Objective To explore the pathological mechanism underlying the exacerbation of ischemic stroke(IS)injury by ovarian aging(OVA),and to elucidate the therapeutic mechanism of curcumin screened based on pathological targets against OVA-aggravated IS injury.Methods Datasets of OVA(GSE81579)and IS(GSE16561)were retrieved from the Gene Expression Omnibus(GEO)database.Core intersecting genes were screened through differential expression analysis,Gene Set Enrichment Analysis(GSEA),and Venn analysis.The Database for Annotation,Visualization and Integrated Discovery(David)was used for Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analyses.Potential traditional Chinese medicines(TCMs)were screened via the Coremine Medicine database,and their active ingredients were further identified through literature review.A middle cerebral artery occlusion/reperfusion(MCAO/R)model was established in female Sprague-Dawley(SD)rats.The rats were randomly divided into five groups:the sham operation(Sham)group,MCAO/R model group,ovariectomy combined with MCAO/R(M&OVA)group,ovariectomy+MCAO/R+curcumin treatment(M&OVA&Cur)group,and ovariectomy+MCAO/R+curcumin+nicotinamide intervention(M&OVA&Cur&Nico)group.Multiple detection methods including neurological function scoring,2,3,5-triphenyltetrazolium chloride(TTC)staining,hematoxylin-eosin(H&E)staining,immunofluorescence assay,transmission electron microscopy(TEM),and Western blot were adopted to verify the protective effect and mechanism of curcumin.Results A total of 80 core intersecting genes were identified between OVA and IS,which were mainly enriched in the inflammatory response,C-X3-C chemokine binding,and NOD-like receptor signaling pathways.Curcuma longa and its active ingredient curcumin were finally screened as the targeted therapeutic agent.In vivo animal experiments demonstrated that rats in the M&OVA group exhibited significantly aggravated cerebral injury compared with those in the sole MCAO/R group.Treatment with curcumin at the dose of 200 mg/kg per day remarkably alleviated neurological deficits,reduced cerebral infarct volume,and inhibited the activation of NLRP3 inflammasome in M&OVA rats.Moreover,the neuroprotective effects of curcumin were markedly abrogated after intervention with nicotinamide,a specific SIRT1 inhibitor.Conclusion Ovarian aging aggravates ischemic stroke injury by activating inflammatory responses.Curcumin can attenuate OVA-aggravated cerebral ischemic injury via regulating the SIRT1-NLRP3 signaling axis,which provides a reliable theoretical basis for the clinical treatment of OVA-complicated ischemic stroke.

Key words: ovarian aging, ischemic stroke, curcumin, NLRP3 inflammasome, SIRT1