Obesity-associated nonalcoholic fatty liver disease (NAFLD) is characterized by excessive intrahepatic lipid accumulation. Despite the increasing prevalence of NAFLD and obesity, the pathogenesis of NAFLD has not yet been clearly elucidated. Prohibitin 1 (PHB1) is mainly expressed in the inner membrane of mitochondria and is known to play an important role in hepatocyte proliferation and lipid metabolism. In this study, we investigated how PHB1 affects lipid metabolism in murine hepatocytes. To reduce the expression of PHB1, Phb1 small interfering RNA was transfected into normal murine hepatocytes (AML12), and the cells were treated with the saturated fatty acid (SFA), palmitic acid (PA), for 24 h. When PHB1 was inhibited, the cell viability decreased by ∼20%, and it was found that it diminished further after PA treatment in both control and peroxisome proliferator-activated receptor gamma (Ppar-γ) knockdown cell groups. Examination of the mRNA expression levels of key enzymes involved in lipid metabolism revealed that PHB1 led to increased stearoyl-coenzyme A desaturase-1 (Scd1) mRNA levels, which leads to an increase in the synthesis of triglycerides (TGs). It also activates the endoplasmic reticulum (ER) stress response through upregulating C/EBP homologous protein (Chop) mRNA levels. PPAR-γ, which has been reported to be upregulated in NAFLD patients, also showed elevated expression. The expression of carnitine palmitoyltransferase 1A, which is involved in the conversion of excess intracellular SFA to fatty acid by catabolism, was downregulated in the PHB1-deficient group. Furthermore, TG synthesis was further promoted by a marked increase in SCD1 mRNA levels, which was further exacerbated by elevated Chop mRNA levels and Ppar-γ disruption. Taken together, PHB1 deficiency led to altered lipid metabolism, resulting in the increased intracellular lipid accumulation and ER stress. These cytotoxic effects were shown to be further exacerbated by excessive PA treatment.
Keywords: lipid metabolism; liver toxicity; palmitic acid; prohibitin 1; saturated fatty acid.