Research Paper Volume 12, Issue 7 pp 6415—6435

Electrical stimulation inhibits Val-boroPro-induced pyroptosis in THP-1 macrophages via sirtuin3 activation to promote autophagy and inhibit ROS generation

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Figure 4. ES inhibits ROS production in VbP-treated THP-1 macrophages. (A) TEM analysis of ultrastructural alterations in THP-1 macrophages. Red arrows indicate cell swelling and nuclear pyknosis (scale bar: 10 μm). (B) DCFH-DA staining showing ROS generation in control macrophages following ES at different voltages (scale bar: 100 μm). (C) DCFH-DA staining showing ROS generation in VbP-treated macrophages following ES at various voltages (scale bar: 100 μm). (D) DCFH-DA staining showing ROS generation in VbP-treated macrophages exposed to NAC and ES (scale bar: 100 μm). (E) Effect of ES, alone or combined with NAC, on NLRP3 immunofluorescence in VbP-treated macrophages (scale bar: 20 μm). (F) ELISA analysis of the effects of NAC and ES on (1) IL-18 and (2) IL-1β secretion by THP-1 macrophages. (G) RT-qPCR analysis of the effects of NAC and ES on pyroptosis-related mRNAs. (H) Western blot analysis of the effects of NAC and ES on pyroptosis-related proteins. n =3; *P<0.05, **P<0.01, and ***P<0.001 vs. control cells; #P<0.05, ##P<0.01, and ###P<0.001 vs. VbP-treated cells.