Optimizing human hepatocyte models for metabolic phenotype and function: effects of treatment with dimethyl sulfoxide (DMSO).
Optimizing human hepatocyte models for metabolic phenotype and function: effects of treatment with dimethyl sulfoxide (DMSO).
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DOI：
10.14814/phy2.12944
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发表时间：
2016-11
影响因子：
2.5
通讯作者：
Tomlinson JW
中科院分区：
文献类型：
--
作者：
Nikolaou N;Green CJ;Gunn PJ;Hodson L;Tomlinson JW
原代人肝细胞被认为是研究肝脂肪酸和葡萄糖代谢的“金标准”细胞模型；然而，它们也有局限性。虽然HepG2细胞系保留了许多原发性肝细胞代谢功能，但它们具有恶性起源和低甘油三酯分泌率。本研究的目的是研究在HepG2细胞培养基中补充二甲亚砜是否会增强代谢功能，从而建立一种改进的体外细胞模型，该模型能很好地概括人原代肝细胞的代谢。HepG2细胞在含1%二甲亚砜的培养基中培养2、4、7、14和21天。测量基因表达、蛋白质水平、细胞内甘油三酯和培养基中甘油三酯、尿素和3 -羟基丁酸浓度。二甲亚砜处理改变了脂质（FAS, ACC1, ACC2, DGAT1, DGAT2， SCD）和葡萄糖（PEPCK, G6Pase）代谢以及肝功能（白蛋白，α - 1 -抗胰蛋白酶，AFP）相关基因的表达。mRNA的变化与蛋白水平的改变是平行的。DMSO处理降低了细胞内甘油三酯含量和乳酸生成，并以时间依赖的方式增加了培养基中甘油三酯和3 -羟基丁酸盐浓度。我们已经证明，在培养基中添加1%二甲亚砜可以改变HepG2细胞的代谢表型，使其向更初级的人肝细胞表型转变。这将增强目前可用的体外模型系统，用于研究与导致疾病的病理过程相关的肝细胞生物学及其对特定治疗干预的反应。
Primary human hepatocytes are considered to be the “gold standard” cellular model for studying hepatic fatty acid and glucose metabolism; however, they come with limitations. Although the HepG2 cell line retains many of the primary hepatocyte metabolic functions they have a malignant origin and low rates of triglyceride secretion. The aim of this study was to investigate whether dimethyl sulfoxide supplementation in the media of HepG2 cells would enhance metabolic functionality leading to the development of an improved in vitro cell model that closely recapitulates primary human hepatocyte metabolism. HepG2 cells were cultured in media containing 1% dimethyl sulfoxide for 2, 4, 7, 14, and 21 days. Gene expression, protein levels, intracellular triglyceride, and media concentrations of triglyceride, urea, and 3‐hydroxybutyrate concentrations were measured. Dimethyl sulfoxide treatment altered the expression of genes involved in lipid (FAS, ACC1, ACC2, DGAT1, DGAT2, SCD) and glucose (PEPCK, G6Pase) metabolism as well as liver functionality (albumin, alpha‐1‐antitrypsin, AFP). mRNA changes were paralleled by alterations at the protein level. DMSO treatment decreased intracellular triglyceride content and lactate production and increased triglyceride and 3‐hydroxybutyrate concentrations in the media in a time‐dependent manner. We have demonstrated that the addition of 1% dimethyl sulfoxide to culture media changes the metabolic phenotype of HepG2 cells toward a more primary human hepatocyte phenotype. This will enhance the currently available in vitro model systems for the study of hepatocyte biology related to pathological processes that contribute to disease and their response to specific therapeutic interventions.