Coenzyme Q₀ Inhibits NLRP3 Inflammasome Activation through Mitophagy Induction in LPS/ATP-Stimulated Macrophages

Figure 7

CoQ₀ inhibits ROS-mediated NLRP3 inflammasome activation through autophagy induction and Nrf2 activation in LPS/ATP-stimulated RAW264.7 macrophages. Cells were pretreated with CoQ₀ (2.5-10 μM), Mito-TEMPO (0.5 mM), or NAC (2 mM) for 1 h and then stimulated with LPS (1 μg/mL) for 5 h followed by ATP (5 mM) for 1 h. (a) Immunofluorescence staining of RAW264.7 cells and the nuclear localization of NLRP3 were visualized by the immunofluorescence method. Cells were stained with DAPI (1 μg/mL) for 5 min and examined by fluorescence microscopy. (b) The expression of pro-IL1β or LC3-I/II proteins was measured by Western blot analysis. (c) Nrf2 knockdown attenuated the protective effects of CoQ₀. Cells were transfected with siRNA that is specific to either Nrf2 or a nonsilencing control. Transfected cells were pretreated with CoQ₀ (2.5-10 μM) for 1 h and then stimulated with LPS (1 μg/mL) for 5 h followed by ATP (5 mM) for 1 h, and the expression of Nrf2 or pro-IL1β proteins in both control and siNrf2 was measured by Western blot analysis.

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