Study on the mechanism of Boschniakia rossica polysaccharide inhibiting the activation of inflammasomes and pyroptosis in J774A.1 macrophages

Abstract

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.

Keywords

• boschnikia rossica
• polysaccharides
• macrophages
• inflammasome
• signaling pathways