Abstract

Non-alcoholic fatty liver disease (NAFLD) is associated with lipid accumulation and lipotoxicity. The main aim of this study is to evaluate the synergistic treatment effect of fish oils (FOs) and chicoric acid (CA) in palmitate (PA)-induced NAFLD HepG2 model. HepG2 cells were pre-treated with palmitate (0.75 mM) for 24 h, and then were exposed to CA, FOs and combination of these chemicals for another 24 h. Gene expression and protein levels were determined using qRT-PCR and western blotting or ELISA analysing, respectively. The combination index (CI) values of FOs and CA in HepG2 cells were calculated according to the Chou-Talalay equation using the CompuSyn software. FOs and CA acid together synergistically reduced lipid accumulation as indicated by decreased oil red O staining (vehicle-treated control: 1 ± 0.1; PA-treated control: 4.7 ± 0.4; PA + CA100: 3.9 ± 0.4; PA + CA200: 2.4 ± 0.3; PA + FOs: 2.7 ± 0.1; PA + CA200 + FOs: 1.5 ± 0.1) and triglyceride (vehicle-treatedcontrol:10 ± 1.2; PA-treated control: 25.8 ± 2.7; PA + CA100: 18.9 ± 2.5; PA + CA200: 14.4 ± 1.8; PA + FOs: 15.2 ± 2.4; PA + CA200 + FOs: 11.9 ± 1.5) levels in PA-treated HepG2 cells. Gene expression and Immunoblotting analysis confirmed the combination effect of FOs and CA in up-regulation of AMPK-mediated PPARα/UCP2 and down-regulation of AMPK-mediated SREBP-1/FAS signalling pathways. Collectively, these results suggest that combining FOs with CA can serve as a potential combination therapy for NAFLD.

摘要

非酒精性脂肪性肝病 (NAFLD) 与脂质积累和脂毒性有关。本研究的主要目的是评估鱼油 (FO) 和菊苣酸 (CA) 在棕榈酸酯 (PA) 诱导的 NAFLD HepG2 模型中的协同治疗效果。HepG2 细胞用棕榈酸酯 (0.75 mM) 预处理 24 小时，然后再暴露于 CA、FO 和这些化学物质的组合 24 小时。分别使用 qRT-PCR 和蛋白质印迹或 ELISA 分析确定基因表达和蛋白质水平。使用 CompuSyn 软件根据 Chou-Talalay 方程计算 HepG2 细胞中 FO 和 CA 的组合指数 (CI) 值。FOs 和 CA 酸一起协同减少脂质积累，如油红 O 染色减少所示（载体处理的对照：1 ± 0.1；PA 处理的对照：4.7 ± 0.4；PA+CA100：3.9±0.4；PA+CA200：2.4±0.3；PA + FO：2.7 ± 0.1；PA + CA200 + FO：1.5 ± 0.1) 和甘油三酯（载体处理的对照：10 ± 1.2；PA 处理的对照：25.8 ± 2.7；PA + CA100：18.9 ± 2.5；PA + CA200：14.4 ± 1.8；PA + FO： 15.2 ± 2.4；PA + CA200 + FO：11.9 ± 1.5) PA 处理的 HepG2 细胞水平。基因表达和免疫印迹分析证实了 FO 和 CA 在上调 AMPK 介导的 PPARα/UCP2 和下调 AMPK 介导的 SREBP-1/FAS 信号通路方面的联合作用。总的来说，这些结果表明将 FOs 与 CA 结合可以作为 NAFLD 的潜在联合疗法。PA + FO：15.2 ± 2.4；PA + CA200 + FO：PA 处理的 HepG2 细胞中的水平为 11.9 ± 1.5)。基因表达和免疫印迹分析证实了 FO 和 CA 在上调 AMPK 介导的 PPARα/UCP2 和下调 AMPK 介导的 SREBP-1/FAS 信号通路方面的联合作用。总的来说，这些结果表明将 FOs 与 CA 结合可以作为 NAFLD 的潜在联合疗法。PA + FO：15.2 ± 2.4；PA + CA200 + FO：PA 处理的 HepG2 细胞中的水平为 11.9 ± 1.5)。基因表达和免疫印迹分析证实了 FO 和 CA 在上调 AMPK 介导的 PPARα/UCP2 和下调 AMPK 介导的 SREBP-1/FAS 信号通路方面的联合作用。总的来说，这些结果表明将 FOs 与 CA 结合可以作为 NAFLD 的潜在联合疗法。