AJTR Copyright © 2009-All rights reserved. Published by e-Century Publishing Corporation, Madison, WI 53711
Am J Transl Res 2011;3(3):284-291

Original Article
Lipotoxicity in HepG2 cells triggered by free fatty acids  

Hong-Rui Yao, Jun Liu, Daniel Plumeri, Yong-Bing Cao, Ting He, Ling Lin, Yu Li, Yuan-Ying Jiang, Ji Li, Jing

Systems Biology Group, Drug Screening Center, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing
210009, Jiangsu Province, China; New Drug Research Center, Second Military Medical University,325 Guo He
Road, Shanghai 200433, China; Department of Pharmacology and Toxicology, School of Medicine and
Biomedical Sciences, University at Buffalo-SUNY, Buffalo, NY, USA.

Received April 27, 2011; Accepted May 10, 2011; Epub May 10, 2011; Published May 15, 2011

Abstract: The goal of this study was to investigate the lipid accumulation and lipotoxicity of free fatty acids (FFAs)
induced in HepG2 cells. HepG2 cells were co-incubated with various concentrations of FFAs for 24h and the
intracellular lipid contents were observed by Oil Red O and Nile Red staining methods. The lipotoxicity of HepG2
cells were then detected by Hoechest 33342/PI, Annexin V-FITC/PI double-staining and
3-(4,5-dimethylthiazol-2-yl)-2,5-di phenyltetrazolium bromide (MTT) experiment tests. The experiments showed a
lipid accumulation and lipotoxicity by increasing FFA concentration gradients. Through cell morphological
observation and quantitative analysis, FFAs have shown to increase in a dose-dependent manner compared with
the control group. The data collected from hoechst 33342/PI, annexin V-FITC/PI double staining and also MTT
experiments showed that cell apoptosis and necrosis significantly increased with increasing FFA concentrations.
Apoptosis was not obvious in the 1 mM FFAs-treated group compared to the other two groups. In a certain
concentration range, FFAs induced intracellular lipid accumulation and lipotoxicity of HepG2 cells in a
dose-dependent manner. (AJTR1104004).

Keywords: Free fatty acids, lipid accumulation, lipotoxicity, HepG2 cells, nonalcoholic fatty liver disease

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Address all correspondence to:
Jing Shang, PhD
Drug Screening Center
China Pharmaceutical University
Nanjing 210009, Jiangsu Province

Yuan-Ying Jiang, PhD
New Drug Research Center
Second Military Medical University
325 Guo He Road, Shanghai 200433
Tel: +86 25 83271043; Fax: +86-25-83271142