Yao Hong-Rui, Liu Jun, Plumeri Daniel, Cao Yong-Bing, He Ting, Lin Ling, Li Yu, Jiang Yuan-Ying, Li Ji, Shang Jing
Am J Transl Res. 2011 May 15;3(3):284-91. Epub 2011 May 10.
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.
本研究的目的是调查游离脂肪酸（FFA）诱导HepG2细胞中的脂质积累和脂毒性。将HepG2细胞与不同浓度的FFA共同孵育24小时，并通过油红O和尼罗红染色方法观察细胞内脂质含量。然后通过Hoechest 33342/PI、膜联蛋白V-FITC/PI双染和3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）实验检测HepG2细胞的脂毒性。实验显示，随着FFA浓度梯度增加，脂质积累和脂毒性增强。通过细胞形态观察和定量分析，与对照组相比，FFA呈剂量依赖性增加。从Hoechest 33342/PI、膜联蛋白V-FITC/PI双染以及MTT实验收集的数据表明，随着FFA浓度增加，细胞凋亡和坏死显著增加。与其他两组相比，1 mM FFA处理组的凋亡不明显。在一定浓度范围内，FFA以剂量依赖性方式诱导HepG2细胞内脂质积累和脂毒性。
