Synergistic Effects of Cilostazol and Probucol on ER Stress-Induced Hepatic Steatosis via Heme Oxygenase-1-Dependent Activation of Mitochondrial Biogenesis
Combinatorial treatment of cilostazol (CZ) and probucol (PB) ameliorates tunicamycin- (TM-) induced hepatosteatosis in vivo. C57BL/6 mice were pretreated with cilostazol (3 mg/kg body weight) and probucol (1 mg/kg body weight) individually or combinatorially once daily for 3 days by intraperitoneal injection. The mice were sacrificed after challenge with tunicamycin (3 mg/kg body weight) for 24 h. Liver tissues were excised and representative liver histology is shown by H&E staining (a). Liver sections were, respectively, from CON (A), TM (B), TM + PB (C), TM + CZ (D), and TM + PB + CZ (E) treated mice. The expression of lipogenesis-related genes SREBP-1c and FAS in liver was measured by real-time RT-PCR (b and c). Triglyceride levels of serum (d), liver tissues (e), HepG2 cells (f) and primary hepatocytes (g), and ALT concentrations (h) were measured. For liver ER stress related proteins, p-eIF2α and CHOP were measured by Western blot (i). The expression of HO-1 and mitochondrial biogenesis-related genes PGC-1α, TFAM, and NRF-1, as well as related proteins PGC-1α, COX III, and COX IV, was measured by real-time RT-PCR (j) and Western blot (k), respectively. Bar graphs, left panels of (i) and (k), are summary data of normalized densitometric ratios. All mice were separated into experimental groups ( mice per group). Quantitative data are expressed as means ± SE; . , , and versus cells without treatment; , , and versus cells treated with tunicamycin; , , and versus cells treated with probucol and tunicamycin; , , and versus cells treated with cilostazol and tunicamycin.