非诺贝特缓解非酒精性脂肪性肝病的体外机制研究  

Therapeutic Effects of Fenofibrate on Nonalcoholic Fatty Liver Disease in vitro and Related Mechanism

作　　者：郭骏[1] 陈玉媛[1] 叶枫[2] 李斌[1] 徐传瑞[1] 

机构地区：[1]华中科技大学同济医学院药学院,武汉430030 [2]华中科技大学同济医学院附属同济医院儿科,武汉430030

出　　处：《华中科技大学学报（医学版）》2017年第3期265-270,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的利用细胞模型研究非诺贝特在缓解非酒精性脂肪性肝病(NAFLD)中的作用及其分子机制。方法首先以0~0.9mmol/L梯度浓度的油酸处理正常肝细胞LO2 48h建立脂肪肝细胞模型;同时,以非诺贝特处理正常细胞或模型组细胞48h,然后用CCK-8试剂盒测定细胞活力,用油红O染色观察细胞内脂滴形成或抑制情况,利用试剂盒检测细胞中的三酰甘油(TG)以及上清液中谷丙转氨酶(ALT)、谷草转氨酶(AST)水平,蛋白质免疫印迹法检测细胞中蛋白激酶B(AKT)、p-AKT、FSAN及Nrf2蛋白水平,并使用2′,7′-二氯荧光黄双乙酸盐(DCFH-DA)检测细胞内的活性氧自由基(ROS)含量。结果油酸浓度超过0.5mmol/L后细胞活力显著降低,非诺贝特可抑制高浓度油酸对细胞的损伤。油酸处理后细胞中出现大量脂滴,细胞内TG含量以及上清液中的ALT、AST水平显著升高,给予非诺贝特处理后细胞中脂滴减少,TG、ALT和AST含量均显著降低。油酸处理后细胞中AKT、p-AKT、FSAN和Nrf2蛋白水平升高,给予非诺贝特可抑制上述蛋白水平的升高。同时,非诺贝特逆转了由油酸导致的细胞内ROS累积。结论非诺贝特可以抑制油酸引起的脂肪积累,并缓解脂肪累积导致的细胞损伤,其机制是通过下调AKT/FSAN信号通路,从而降低ROS水平,进而减少ROS导致的细胞损伤。Objective To investigate the role of fenofibrate in treating non-alcoholic fatty liver disease（NAFLD）in cellular model and the related mechanism.Methods NAFLD cell model was established by treating LO2 cells with oleic acid（from 0to0.9mmol/L）.The cells were treated with fenofibrate（from 0to 20 mmol/L）for 48 h.CCK-8kit was used to determine the LO2 cell viability with or without treatment.Oil Red O staining was used to observe lipid droplets in LO2 cells microscopically.Lab-used experiment kits were used to assess TG level in cells and ALT,AST levels in cell supernatants.Protein levels of AKT,p-AKT,FSAN and Nrf2 were detected using Western blotting.DCFH-DA was used to detect reactive oxygen species（ROS）in cells.Results Oleic acid with concentration over 0.5mmol/L significantly inhibited cell viability.Fenofibrate significantly reversed cytotoxicity induced by high concentration of oleic acid.Lipid droplets and TG within LO2 cells,and ALT/AST in cell supernatant were increased by treatment with oleic acid.Fenofibrate reversed the increase of lipid drops,TG,ALT and AST induced by oleic acid.Oleic acid up-regulated the levels of AKT,p-AKT,FSAN and Nrf2,while fenofibrate reversed the up-regulation of those proteins.Fenofibrate reduced the oleic acid-induced accumlation of ROS in cells.Conclusion Fenofibrate can inhibit lipid accumulation and alleviate cell damage induced by oleic acid through inhibiting AKT/FSAN pathways and reducing ROS levels.

关 键 词：非酒精性脂肪性肝病 非诺贝特 蛋白激酶B(AKT) 活性氧自由基 

分 类 号：R575.5[医药卫生—消化系统]