文章摘要
林霖 盛夏 吉猛伍开明林勇曾欣.油酸体外诱导细胞脂肪变性对脂质代谢及炎症的影响[J].,2016,16(25):4810-4814
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 inHepG2 Cells
  
DOI:
中文关键词: 人肝癌细胞株  脂肪变性  脂质代谢  炎症
英文关键词: Human Hepatoma Cell Line  Steatosis  Lipid Metabolism  Inflammation
基金项目: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)
作者单位
林霖 盛夏 吉猛伍开明林勇曾欣 第二军医大学长征医院消化内科 
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中文摘要:
      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 mM)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 mM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 mM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。
英文摘要:
      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 mM 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.Conclusion:Steatosis model of hepatocyte could be successfully established by treating HepG2 cells with 0.5 mMoleic acid and this model may provide a new method to explore the mechanismof NAFLD in vitro.
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