Effect of Rab18 on liver injury and lipid accumulation by regulating perilipin 2 and peroxisome proliferator‐activated receptor gamma in non‐alcoholic fatty liver disease

Abstract

Background and AimNonalcoholic fatty liver disease (NAFLD) is currently one of the most common chronic liver diseases worldwide, characterized by the presence of lipid droplets. Rab18 is an important lipid droplet protein; however, its effects and mechanisms of action on NAFLD remain unclear.MethodsFree fatty acid‐stimulated AML‐12 cells and high‐fat diet (HFD)‐fed mice were used as NAFLD models. Lentiviruses overexpressing Rab18 (Rab18‐OE) or knockdown (Rab18‐KD) were used to generate stable cell lines for genetic analysis. Blood serum levels of alanine aminotransferase, aspartate aminotransferase, total cholesterol, triglycerides, high‐density lipoprotein cholesterol, low‐density lipoprotein cholesterol, glucose, and leptin were measured using a biochemical autoanalyzer. Hematoxylin and eosin staining was performed to detect pathological damage to the liver. Lipid accumulation in the cells was assessed by Oil Red O staining. Target expression was measured using qPCR, western blotting, and immunocytochemistry.ResultsRab18 mRNA and protein expression levels increased in free fatty acid‐stimulated AML‐12 cells and the livers of HFD‐fed mice. Rab18‐OE increased lipid accumulation in vitro, which was attenuated by Rab18‐KD. In vivo, Rab18‐OE augmented liver pathological damage, serum alanine aminotransferase/aspartate aminotransferase activity, and triglyceride, total cholesterol, and low‐density lipoprotein levels, whereas Rab18‐KD decreased these indicators. Rab18‐KD also downregulated blood glucose levels in HFD‐fed mice. Mechanistically, Rab18‐OE and Rab18‐KD regulated the mRNA and protein expression levels of perilipin 2 (PLIN2) and peroxisome proliferator‐activated receptor gamma (PPARγ) in vitro and in vivo, respectively. Immunocytochemistry revealed that Rab18 colocalized with PLIN2 and PPARγ in AML‐12 cells.ConclusionRab18 expression was elevated in vitro and in vivo in the NAFLD mouse model. Rab18 regulates PLIN2 and PPARγ expression to exaggerate liver injury and lipid accumulation in patients with NAFLD. Thus, Rab18 may be a crucial protein in this disease and a potential therapeutic target.