Omanovic Kolaric Tea, Kizivat Tomislav, Mihaljevic Vjera, Zjalic Milorad, Bilic-Curcic Ines, Kuna Lucija, Smolic Robert, Vcev Aleksandar, Wu George Y, Smolic Martina
Department of Pharmacology and Biochemistry, Faculty of Dental Medicine and Health Osijek, Crkvena 21, 31000 Osijek, Croatia.
Department of Pharmacology, Faculty of Medicine Osijek, J. Huttlera 4, 31000 Osijek, Croatia.
Curr Issues Mol Biol. 2022 Aug 2;44(8):3465-3480. doi: 10.3390/cimb44080239.
(1) Background: With the aging of the population and polypharmacy encountered in the elderly, drug-induced steatosis (DIS) has become frequent cause of non-alcoholic steatosis (NAS). Indeed, NAS and DIS may co-exist, making the ability to distinguish between the entities ever more important. The aim of our study was to study cell culture models of NAS and DIS and determine the effects of liraglutide (LIRA) in those models. (2) Methods: Huh7 cells were treated with oleic acid (OA), or amiodarone (AMD) to establish models of NAS and DIS, respectively. Cells were treated with LIRA and cell viability was assessed by MTT, lipid accumulation by Oil-Red-O staining and triglyceride assay, and intracellular signals involved in hepatosteatosis were quantitated by RT-PCR. (3) Results: After exposure to various OA and AMD concentrations, those that achieved 80% of cells viabilities were used in further experiments to establish NAS and DIS models using 0.5 mM OA and 20 µM AMD, respectively. In both models, LIRA increased cell viability (p < 0.01). Lipid accumulation was increased in both models, with microsteatotic pattern in DIS, and macrosteatotic pattern in NAS which corresponds to greater triglyceride accumulation in latter. LIRA ameliorated these changes (p < 0.001), and downregulated expression of lipogenic ACSL1, PPARγ, and SREBP-1c pathways in the liver (p < 0.01) (4) Conclusions: LIRA ameliorates hepatocyte steatosis in Huh7 cell culture models of NAS and DIS.
（1）背景：随着人口老龄化以及老年人中多重用药情况的出现，药物性脂肪变性（DIS）已成为非酒精性脂肪变性（NAS）的常见原因。事实上，NAS和DIS可能同时存在，因此区分这两种情况的能力变得愈发重要。我们研究的目的是研究NAS和DIS的细胞培养模型，并确定利拉鲁肽（LIRA）在这些模型中的作用。（2）方法：分别用油酸（OA）或胺碘酮（AMD）处理Huh7细胞，以建立NAS和DIS模型。用LIRA处理细胞，通过MTT法评估细胞活力，用油红O染色和甘油三酯测定法评估脂质积累，并通过RT-PCR定量肝脂肪变性相关的细胞内信号。（3）结果：在暴露于不同浓度的OA和AMD后，选择细胞活力达到80%的浓度用于进一步实验，分别用0.5 mM OA和20 µM AMD建立NAS和DIS模型。在两种模型中，LIRA均提高了细胞活力（p < 0.01）。两种模型中的脂质积累均增加，DIS呈现微脂肪变性模式，NAS呈现大脂肪变性模式，后者对应的甘油三酯积累更多。LIRA改善了这些变化（p < 0.001），并下调了肝脏中脂肪生成相关的ACSL1、PPARγ和SREBP-1c通路的表达（p < 0.01）。（4）结论：LIRA改善了NAS和DIS的Huh7细胞培养模型中的肝细胞脂肪变性。
