油酸体外诱导细胞脂肪变性对脂质代谢及炎症的影响  

The Effect of Oleic Acid on Induction of Steatosis and Inflammation in HepG2 Cells

作　　者：林霖[1] 盛夏[1] 吉猛[1] 伍开明[1] 林勇[1] 曾欣[1] 

机构地区：[1]第二军医大学长征医院消化内科,上海200003

出　　处：《现代生物医学进展》2016年第25期4810-4814,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81370526);上海市卫生系统优秀人才培养(新优青)计划(XYQ2013103)

摘　　要：目的:利用不同浓度油酸处理人肝肿瘤细胞株HepG2,选取最佳浓度油酸后,观察最佳浓度的油酸对细胞脂质合成及代谢、肝细胞功能相关基因表达的影响及NF-κB、IL-6通路的变化,建立稳定高效的肝细胞脂肪变性体外研究模型。方法:将HepG2细胞用不同浓度油酸(0、0.5、0.75、1.25、1.5 m M)处理,油红O染色选取最佳浓度后,利用Realtime RT-PCR检测细胞内脂质合成、代谢及肝细胞功能相关基因表达情况,Western blot检测细胞内TLR4-TNFα-NF-κB及IL-6通路活性。结果:利用0.5 m M浓度油酸处理HepG2后,细胞未见明显死亡现象,细胞内脂质合成指标表达增多、代谢指标表达也明显增加,肝细胞功能相关基因表达下调,TLR4-TNFα-NF-κB、IL-6通路激活。结论:0.5 m M浓度油酸处理细胞可诱导HepG2脂肪变性,利用最佳浓度油酸处理细胞可作为NAFLD体外研究的细胞模型。Objective： To observe the effect of oleic acid on lipid synthesis and metabolism, expression of liver function related genes and inflammatory signal pathways in hepatoma cell line HepG2, and establish the model of hepatocyte steatosis in vitro. Methods：HepG2 cells were treated by different concentrations of oleic acid. Lipid droplets were observed through oil red O staining. Realtime RT-PCR was used to detect the liver function related genes and genes involving in lipid metabolism. Western blot was carried out to measure the activity of inflammatory pathways NF-κB signal pathway and the protein expression of IL-6, one of the most important inflammatory cytokines in NASH. Results： We found the size and number of lipid droplets in HepG2 cells were increased remarkedly after treated with 0.5 m M oleic acid. Oleic acid treatment significantly induced the expression of genes involving in lipid synthesis and metabolism, and reduced the expression of liver function related genes. In addition, Western blot analysis comfirmed the inflammatory pathways NF-κB signal pathway was activated and the protein level of IL-6 was elevated in HepG2 cells after oleic acid delivery. Conclusions： Steatosis model of hepatocyte could be successfully established by treating HepG2 cells with 0.5 m M oleic acid and this model may provide a new method to explore the mechanism of NAFLD in vitro.

关 键 词：人肝癌细胞株 脂肪变性 脂质代谢 炎症 

分 类 号：R-33[医药卫生] R735.7