Fig. 4.

Berberine reduced SIRT3 induction and preserved MPC1 acetylation in the liver. (a): Acetylated-Lysine protein expression in the liver mitochondria. (b): Acetylation in precipitated MPC1 in primary mouse hepatocytes incubated with PA for 24 h. (c–d): SIRT3 gene expression in the liver of HFD-fed mice and PA-treated hepatocytes. (e): SIRT3 protein expression in primary hepatocytes in the presence of 10 mM pyruvate for 8 h. (f): Western blot examination of SIRT3 in precipitated MPC1 protein in hepatocytes incubated with pyruvate for 8 h. (g): Confocal image of mitochondrial MPC1 and SIRT3 colocalization in hepatocytes (Blue: MPC1; Green: SIRT3; Red: MitoTracker Red cMXRos. Scale bars: 5 μm), The view is one of five independent experiments. (h): Knockdown efficiency of three SIRT3 siRNAs transfected in HepG2 cells. (i): MPC1 protein expression in HepG2 with SIRT3 knockdown when cultured with pyruvate (10 mM) for 8 h. (j): Acetylation of MPC1 expression in HepG2 cells overexpressed with SIRT3 when cultured with pyruvate (10 mM) for 8 h. (BBR, berberine; Met, metformin; NAM, nicotinamide; PA, palmitate). Data were expressed as the mean ± SD (n = 5–6). ^⁎p < 0.05 vs. HFD mice, PA or pyruvate-treated cells; ^#p < 0.05 vs. chow-fed diet mice or untreated cells.