High-throughput RNA sequencing identifies hub genes and anti-inflammatory effects of β-caryophyllene in cerebral ischemia-reperfusion via p38MAPK/NF-κB modulation

β-caryophyllene (BCP) has been shown to alleviate neurological deficits in rats with cerebral ischemia-reperfusion injury (CIRI) induced by middle cerebral artery occlusion (MCAO). However, its molecular targets remain unclear. In this study, transcriptome analysis was conducted to identify BCP-responsive genes and potential therapeutic pathways. RNA sequencing revealed that BCP downregulated genes upregulated by CIRI, particularly those involved in extracellular matrix organization, leukocyte migration, angiogenesis regulation, and reactive oxygen species metabolism. KEGG analysis indicated enrichment in the MAPK, NF-κB, and HIF-1 signaling pathways. Male SD rats were randomly divided into Sham, CIRI, CIRI+BCP (306 mg/kg), CIRI+BCP+Diprovocim, and Diprovocim-only groups. After 1.5 h of ischemia followed by 24 h of reperfusion, neurological scores, infarct volume, MAPK/NF-κB protein expression (by Western blot), hippocampal neuron damage (by HE staining), proinflammatory cytokine levels (TNF-α and IL-1β via ELISA), and oxidative stress markers (SOD and MDA) were evaluated. BCP treatment significantly reduced infarct volume, improved neurological function, downregulated MAPK and NF-κB expression, suppressed TNF-α and IL-1β release, and reduced neuronal death (all P < 0.05). These effects were partially reversed by the MAPK agonist Diprovocim, suggesting the involvement of the p38MAPK/NF-κB pathway in BCP's protective mechanism. In conclusion, BCP confers neuroprotection in CIRI by modulating key signaling pathways, particularly p38MAPK/NF-κB, highlighting its therapeutic potential in ischemic stroke.