Indole-3-acetic acid alleviates Cryptococcus neoformans-induced pyroptosis in cerebral microvascular endothelial cells by regulating stress granule-mediated NLRP3 inflammasome activation
CHEN Jingyu
ZOU Jinhu
ZHOU Bingliang
GAO Xuefeng
HUANG Pengwei
CAO Hong
Abstract：Objective To investigate whether indole-3-acetic acid(IAA)alleviates Cryptococcus neoformans(Cn)-induced pyroptosis in cerebral microvascular endothelial cells by modulating stress granules(SGs)formation and the NLRP3 inflammasome.Methods In vitro cultured cerebral microvascular endothelial cells were pretreated with different concentrations of IAA before Cn infection(10⁷/mL),and the changes in cellular expresisons of G3BP1,DDX3X,NLRP3 and pyroptosis-related proteins,cytokines,and cell viability were deceted using Western blotting,immunofluorescence staining,ELISA,and CCK-8 assay.In the animal experiment,C57BL/6 mice with cyclophosphamide-induced immunosuppression were pretreated with saline or IAA gavage for 7 days before intravenous Cn injection.The changes in blood-brain barrier(BBB)integrity of the mice was assessed with Evans blue assay,and the brain cortical tissues were analyzed for changes in protein expressions.Results Cn infection significantly downregulated G3BP1 expression and upregulated the expressions of DDX3X and NLRP3 in cultured cerebral microvascular endothelial cells.IAA intervention not only restored normal expressions of G3BP1,DDX3X,and NLRP3,but also effectively suppressed the activation of pyroptosis-related proteins,including NT-GSDMD/GSDMD and P20/caspase-1,and reduced the release of IL-18 and IL-1β.IAA treatment also inhibited the translocation of DDX3X to NLRP3 induced by Cn infection and promoted the binding between DDX3X and G3BP1.In Cn-infected C57BL/6 mice,IAA treatment significantly alleviated BBB injury,decreased the expression of ZO-1 in the cerebral cortex,and effectively ameliorated abnormal expressions of VEGFR2,G3BP1,DDX3X,NLRP3,NT-GSDMD/GSDMD and P20/caspase-1.Conclusion IAA effectively alleviates Cn infection-induced pyroptosis of cerebral microvascular endothelial cells by modulating the formation of SGs and activating the NLRP3 inflammasome.
Keywords：Cryptococcus neoformanspyroptosisindole-3-acetic acidstress granulesNLRP3 inflammasome
Publication Date：2025-12-20
Online Publishing Date：2025-12-31(First online date of this platform, not the publication date of the document)
Pages：11( 2679-2689 )