A Cell-Based Assay to Investigate Hypolipidemic Effects of Nonalcoholic Fatty Liver Disease Therapeutics. (March 2018)

• Record Type: Journal Article
• Authors: Dave, Tapan; Tilles, Arno William; Vemula, Muralikrishna
• Is Part Of: SLAS discovery. Volume 23:Number 3(2018)
• Page Start: 274
• Page End: 282
• Publication Date: 2018-03
• Subjects: fatty liver -- hypolipidemic -- NAFLD -- cell-based assays -- fluorescence methods; Drugs -- Analysis -- Periodicals; Drugs -- Testing -- Periodicals; Biomolecules -- Analysis -- Periodicals; Biomolecules -- Analysis; Drugs -- Analysis; Drugs -- Testing; Drug Evaluation, Preclinical; Molecular Biology -- methods; Periodicals; Periodicals; 615.1
• DOI: 10.1177/2472555217741077
• Languages: English
• ISSNs: 2472-5552

Abstract

In the recent past, there has been a growing interest in developing nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) therapeutics. As a result, a need for in vitro cell models of human hepatic steatosis and high-throughput assays to measure intracellular lipid levels has arisen. To address this growing need, we optimized the conditions based on the current literature to fatten HepG2 hepatocytes by adding a mixture of saturated and unsaturated fatty acids (oleate/palmitate, 2:1 molar ratio) without inducing any overt cytotoxicity. Our results indicate that hepatocytes fatten in a concentration- (0.75–1.5 mM of fatty acids) and time-dependent manner, with a substantial increase in intracellular lipid levels seen within 6 h. Additionally, a method to quantify lipid levels in cells using a fluorescent reagent that is more sensitive than that in conventional assays and adaptable for high-throughput screening is presented. Lastly, the utility of the in vitro cell model and an assay based on AdipoRed to measure hypolipidemic effects of therapeutic drugs is demonstrated using fenofibrate, a molecule that was previously shown to lower lipid levels in the liver.