OBJECTIVE To investigate the protective effects and potential mechanisms of Acorus tatarinowii volatile oil and its main components (α-asarone, β-asarone, and methyl eugenol) on inflammation and pyroptosis in lipopolysaccharide (LPS)-induced brain microvascular endothelial cells (bEnd. 3).

METHODS An in vitro inflammation model was established using LPS-stimulated bEnd. 3 cells. The LPS-stimulated bEnd.3 cells were then treated with Acorus tatarinowii volatile oil and its main active components (α-asarone, β-asarone, and methyl eugenol) to investigate their effects on inflammation and pyroptosis. Network pharmacology and molecular docking analysis were used to identify potential targets of methyl eugenol. The STAT3 inhibitor Stattic was used to further validate the role of the signaling pathway.

RESULTS The volatile oil of Acorus tatarinowii and its main active ingredient, methyl eugenol, both exhibited significant anti-inflammatory and anti-pyroptosis effects, effectively inhibiting inflammation and pyroptosis. Network pharmacology analysis suggested that the JAK2/STAT3 signaling pathway might be a potential target of methyl eugenol, and molecular docking experiments further confirmed that methyl eugenol had a strong binding affinity for both JAK2 and STAT3 proteins. In the LPS-induced bEnd. 3 cell model, both Acorus tatarinowii volatile oil and methyl eugenol significantly reduced the phosphorylation level of STAT3; simultaneously, the STAT3-specific inhibitor Stattic also inhibited the expression of the pyroptosis marker protein cleaved caspase-1, further validating the key role of this pathway in regulating pyroptosis.

CONCLUSION Methyl eugenol can significantly inhibit LPS-induced pyroptosis and inflammatory responses in bEnd. 3 cells by targeting the JAK2/STAT3 signaling pathway. The findings offer a new strategy for the prevention and treatment of neuroinflammation-related diseases.