title: Low simvastatin concentrations reduce oleic acid-induced steatosis in HepG2cells: Anin vitromodel of non-alcoholic fatty liver disease.
abstract: Non-alcoholic fatty liver disease (NAFLD) is an inflammatory condition caused by hepatic lipid accumulation that is associated with insulin resistance, diabetes and metabolic syndrome. Although statins should be used with caution in liver diseases, they are increasingly investigated as a possible treatment for NAFLD. The present study recreated anin vitromodel of NAFLD using HepG2cells exposed to oleic acid (OA), which was used to quantify OA-induced lipid accumulation in HepG2cells treated with various concentrations of simvastatin. In addition, the effect of simvastatin on HepG2cell morphology and microparticle generation as a marker of cell apoptosis was assessed. OA-induced lipid accumulation was quantified by Oil Red O staining and extraction for optical density determination. Stained lipid droplets were visualized using phase contrast microscopy. Furthermore, HepG2cell-derived microparticles were counted by flow cytometry subsequent to staining for Annexin V. HepG2cells treated with 0-1 mM OA showed dose-dependent lipid accumulation. Treatment of HepG2cells with increasing concentrations of simvastatin followed by treatment with 1 mM OA showed that low simvastatin concentrations (4-10 µM) were able to reduce lipid accumulation by ~40%, whereas high simvastatin concentrations (20 and 30 µM) induced apoptotic changes in cell morphology and increased the production of Annexin V+microparticles. This suggests that low simvastatin doses may have a role in preventing NAFLD. However, further investigations are required to confirm this actionin vivoand to determine the underlying mechanism by which simvastatin reduces hepatic steatosis.
authors.affiliation: Clinical Pharmacy Department, Faculty of Pharmacy, Jordan University of Science and Technology, Irbid 22110, Jordan.
authors.forename: Mohammad J
authors.initials: MJ
authors.lastname: Alkhatatbeh
authors.affiliation: Hunter Haematology Research Group, Calvary Mater Newcastle Hospital, Waratah, NSW 2298, Australia.
authors.forename: Lisa F
authors.initials: LF
authors.lastname: Lincz
authors.affiliation: School of Environmental and Life Sciences, University of Newcastle, Ourimbah, NSW 2258, Australia.
authors.forename: Rick F
authors.initials: RF
authors.lastname: Thorne
keywords: microparticles; non-alcoholic fatty liver disease; oleic acid; simvastatin; steatosis