[关键词]
[摘要]
目的 研究高浓度(>100 μmol/L)绿原酸(CGA)对正常及非酒精性脂肪肝(NAFLD)状态下L02肝细胞脂质堆积与氧化应激的影响。方法 用666μmol/L油酸(OA)-333 μmol/L棕榈酸(PA)处理人正常肝细胞系L02 24 h构建体外肝细胞NAFLD模型。正常及模型细胞经CGA(0.5、1、2 mmol/L)处理24 h后,油红O染色检测胞内脂滴的量,定量PCR及Western blotting检测细胞SREBP-1C、PNPLA3和CYP2E1的mRNA和蛋白表达量,荧光光度法检测胞内活性氧(ROS)的量。结果 正常及NAFLD模型L02细胞经CGA处理后,均浓度依赖性提高了胞内脂滴与ROS的量,以及SREBP-1C、PNPLA3和CYP2E1的mRNA与蛋白表达水平。结论 高浓度CGA(0.5、1、2 mmol/L)处理能促进正常及NAFLD模型L02细胞的脂堆积与氧化应激损伤。
[Key word]
[Abstract]
Objective To investigate the effects of chlorogenic acid (CGA) at high concentration (> 100 μmol/L) on the lipid accumulation and oxidative stress in L02 cells under normal and non alcoholic liver disease (NAFLD) status induced by oleic acid (OA) and palmic acid (PA) treatment. Methods L02 cells were treatment by OA (666 μmol/L)-PA (333 μmol/L) for 24 h to induce intracellular steatosis. After normal and OA-PA treated cells were treated by CGA (0, 0.5, 1, and 2 mmol/L) for 24 h, the intracellular contents of lipid droplets and reactive oxygen species (ROS) were determined by Oid red staining and fluorospectrophotometry, respectively. And the mRNA and protein expression levels of SREBP-1C, PNPLA3, and CYP2E1 were detected by Real-time PCR and Western blotting, respectively. Results CGA treatment dose-dependently increased the levels of intracellular lipid droplets and ROS, as well as the mRNA and protein expression levels of SREBP-1C, PNPLA3, and CYP2E1 in normal and OA-PA treated L02 cells. Conclusion CGA treatment at high concentration can accelerate the lipid accumulation and oxidative stress injury in L02 cells under normal and NAFLD status.
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[基金项目]
湖北省技术创新专项重大项目(2016ACA140)