Figure 6.

Inhibition of mitophagy by cyclosporin (CsA) blunted the protective effect of Hesperetin (Hst) against palmitic (PA)-triggered NLRP3 inflammasome activation and mitochondrial dysfunction in HepG2 cells. Cells were pretreated with or without CsA (5 μM) for 4 h in the absence or presence of 40 μM of Hst before addition of PA (400 μM). (A) IL-1β production determined by ELISA assay. (B) Representative immunoblots and (C) quantification of NLRP3 and caspase-1. α-Tubulin was used as a loading control. Cells were loaded with JC-1 (mitochondrial membrane potential fluorescent probe, 2 μM) or with mitoSOX (red, mitochondrial ROS indicator, 1 μM), and analyzed by confocal microscope. Representative images and quantification of (D,E) mitochondrial membrane potential and (F,G) mitochondrial ROS. Nuclei were counterstained with Hoechst 33342 (blue). Data are presented as mean ± SEM of n = 3 independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 (unpaired two-tailed student’s t test). Scale bars are 10 μm.