Atractylenolide III alleviates inflammation in cerebral ischemia/reperfusion injury by modulating the PI3K/Akt/NF-κB signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Atractylodes macrocephala, traditionally recorded in classical formulas such as 'Banxia Baizhu Tianma Tang' and 'Xiaoxuming Tang', has been used in traditional medicine for conditions described as 'fēng tán zǔ luò xíng' and 'bì zǔ jīng luò xíng', which are considered related to cerebrovascular disorders. Atractylenolide Ⅲ (ATL Ⅲ), a typical sesquiterpene lactone derived from A. macrocephala, has been reported to exert neuroprotective effects through antioxidant and anti-inflammatory activities. Nevertheless, its specific role and underlying mechanisms in cerebral ischemia reperfusion injury (CIRI) remain to be clarified.

AIM OF THE STUDY: The present study was designed to test the hypothesis that ATL Ⅲ ameliorates CIRI primarily by suppressing inflammation through the PI3K/Akt/NF-κB signaling pathway, which was evaluated in both middle cerebral artery occlusion (MCAO) and oxygen-glucose deprivation/reoxygenation (OGD/R) models.

MATERIALS AND METHODS: In this study, models of MCAO and OGD/R were established to explore its effect of ATL Ⅲ on CIRI. Thereafter, the therapeutic mechanism of ATL Ⅲ via transcriptomics, molecular docking, enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, immunofluorescence, Western blot analysis.

RESULTS: ATL Ⅲ treatment significantly reduced infarct volume, neurological deficits, and pro-inflammatory cytokine release, while preserving blood-brain barrier (BBB) integrity in MCAO mice. In OGD/R-exposed PC12 cells, ATL Ⅲ attenuated oxidative stress, inhibited apoptosis, and decreased inflammatory mediator production. Transcriptomic analysis revealed several significantly enriched pathways following ATL Ⅲ treatment, among which the PI3K/Akt/NF-κB signaling pathway was prominent and therefore guided our mechanistic focus. Molecular docking supported the binding of ATL Ⅲ to key pathway proteins, and inhibition of PI3K with LY294002 attenuated the protective effects of ATL Ⅲ, confirming the central role of this pathway.

CONCLUSION: ATL Ⅲ may inhibit inflammation in CIRI, potentially through regulation of the PI3K/Akt/NF-κB signaling pathway, highlighting its promise as a candidate compound for further preclinical investigation in ischemic stroke.