Fig. 5

Pretreatment with ROS inhibitor suppresses Hcy-induced macrophage pyroptosis. Macrophages were pretreated with ROS inhibitor (NAC, 5 mM) for 1 h, and then the cells were incubated with Hcy (10 mM) for 24 h. A JC-1 assay was used to examine the mitochondrial membrane potential. The scale bars correspond to 100 μm. B Intracellular ROS level was detected using a DCFH-DA probe (green). Mitochondrial ROS level was detected by Mito-Sox Staining (red). The scale bars correspond to 100 μm. C Western blot was used to evaluate the pyroptosis-related proteins level in different groups. NAC pretreatment suppressed the upregulation of pyroptosis-associated protein levels in the presence of Hcy. D–G Quantitative analysis of pyroptosis-associated protein expression. H LDH assay was used to evaluate the cell membrane integrity. I The levels of ATP were measured by commercial kits after Hcy treated. The data are shown as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001