Clofibrate Attenuates ROS Production by Lipid Overload in Cultured Rat Hepatoma Cells

• Published in: Journal of pharmacy & pharmaceutical sciences Vol. 20; pp. 239 - 251
• Main Authors: Chen, Yufei, Li, Wei, Wang, Guqi, Burczynski, Frank J
• Format: Journal Article
• Language: English
• Published: Canada Frontiers Media S.A 01-01-2017
• Subjects: Animals; Cell Proliferation - drug effects; Cell Survival - drug effects; Clofibrate - pharmacology; Dose-Response Relationship, Drug; Fatty Acid-Binding Proteins - antagonists & inhibitors; Fatty Acid-Binding Proteins - biosynthesis; Fatty Acid-Binding Proteins - genetics; Fatty Acids - pharmacology; Lipids - administration & dosage; Lipids - chemistry; Lipids - pharmacology; Particle Size; Rats; Reactive Oxygen Species - antagonists & inhibitors; Reactive Oxygen Species - metabolism; RNA, Small Interfering - pharmacology; Structure-Activity Relationship; Tumor Cells, Cultured
• ISSN: 1482-1826; 1482-1826
• DOI: 10.18433/J3TS70

To investigate the effect of clofibrate on inducing liver fatty acid binding protein (FABP1) following a high-fat load in a hepatocyte cell culture model.
Rat hepatoma cells (CRL-1548) were treated with a fatty acid (FA) mixture consisting of oleate:palmitate (2:1) in the presence of 3% albumin. Cells were treated with 0, 0.5, 1, 2, or 3 mM FA for 24 and 48 hr, or further treated with 500 µM clofibrate (CLO) to induce FABP1 levels. Cytotoxicity was determined using the WST-1 assay. Intracellular lipid droplets were quantitated following staining with Nile Red. Dichlorofluorescein (DCF) was used to assess the extent of intracellular reactive oxygen species (ROS).
Cell viability decreased (p < 0.01) with an increase in lipid concentration. Intracellular lipid droplets accumulated significantly (p < 0.001) with an increase in long-chain fatty acid load, which was associated with a statistical increase (p < 0.05) in ROS levels. Early clofibrate treatment showed significant increases in intracellular FABP1 levels with significant decreases in ROS levels (p < 0.05). Silencing FABP1 expression using siRNA revealed that FABP1 was the main contributor for the observed intracellular ROS clearance.
Characteristic cellular damage resulted from released ROS following a high fat load to hepatoma cells. The damage was attenuated through early treatment with clofibrate, which may act as a hepatoprotectant by inducing FABP1 expression and in this manner, suppress intracellular ROS levels. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.