Study on the mechanism of Boschniakia rossica polysaccharide inhibiting the activation of inflammasomes and pyroptosis in J774A.1 macrophages
Objective:This study aimed to investigate the inhibitory effect of Boschniakia rossica polysaccharides(BRPS)on the activation of inflammasome and pyroptosis of J774A.1 macrophages induced by lipopolysaccharide(LPS)and adenosine triphosphate(ATP).Methods:The nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome model was established by stimulating J774A.1 macrophages with LPS and ATP.Cell viability was detected using the cell counting kit-8,the release of lactate dehydrogenase(LDH)was tested with the microplate method,the cellular level of NLRP3 and cell membrane damage were detected using the immunofluorescence staining method,and the secretion of interleukin-1β(IL-1β)was measured by the enzyme-linked immunosorbent assay.The protein expressions of NLRP3,caspase-1,apoptosis associated speck-like protein containing caspase recruitment domains(ASC),IL-1β,gasdermin D(GSDMD),as well as the nuclear factors-κB(NF-κB)and mitogen activated protein kinase(MAPK)was determined with the western blotting method.Results:BRPS could increase the cell viability,reduce the release of LDH in the culture medium,alleviate the membrane damage of macrophages,down-regulate the protein expression of NLRP3,up-regulate the level of caspase-1 precursor.and down-regulate the levels of IL-1β and GSDMD N-terminal active fragment in the model group.In addition,BRPS suppressed the nuclear translocation and phosphorylation of NF-κB,as well as the phosphorylation of extracellular signal-regulated protein kinase,c-Jun amino terminal kinase,and p38 MAPK.Conclusion:BRPS inhibit LPS/ATP-induced activation of macrophage inflammasome and pyroptosis in mouse J774A.1 cells through regulating the NF-κB and MAPK signaling pathways.
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