Villalva-Pérez José Manuel, Ramírez-Vargas Marco Antonio, Serafín-Fabían Jesús Isimar, Ramírez Mónica, Elena Moreno-Godínez Ma, Espinoza-Rojo Mónica, Flores-Alfaro Eugenia
Laboratorio de Investigación en Epidemiología Clínica y Molecular, Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Guerrero, Av. Lázaro Cárdenas s/n, Zip: 39089, Chilpancingo, GRO, Mexico.
Laboratorio de Toxicología y Salud Ambiental, Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Guerrero, Zip: 39087, Chilpancingo, GRO, Mexico.
Cytotechnology. 2020 Aug;72(4):499-511. doi: 10.1007/s10616-020-00398-4. Epub 2020 May 14.
Liver-specific insulin resistance is associated with the development of the main challenges in metabolism, resulting in dyslipidemia, hyperinsulinemia and hyperglycemia. In vitro models developed for researching hepatic insulin resistance are limited and employed cell lines without similar characteristics to primary human hepatocytes. The Huh7 cell line has been established as a model with similar characteristics to primary human hepatocytes. In addition, it has been identified in the Huh7 cell line that infection with the hepatitis C virus induces insulin resistance. Therefore, we analyzed the induction of insulin resistance (IR) in the Huh7 cell line using an overdosage of insulin and treatment with metformin for its reversal, with the purpose of establishing an insulin resistance model useful for metabolic and pharmacological studies. Insulin-resistant Huh7 (Huh7-IR) showed a reduction in Glut2, glycogen levels, and glucose uptake stimulated by insulin or tyrosine phosphorylation from the β-fraction of insulin receptor post-insulin stimulation, with an increase of glucose production and lipid intracellular content. These biomarkers are frequently observed in insulin-resistant hepatic cells. Moreover, treatment of Huh7-IR with 0.5, 1 or 2 mM of metformin by 24 h decreased the biomarkers associated with an insulin-resistant state. These results suggest that Huh7-IR could be used as an in vitro system to research hepatic insulin resistance in metabolic and pharmacological studies.
肝脏特异性胰岛素抵抗与代谢方面的主要挑战的发展相关，会导致血脂异常、高胰岛素血症和高血糖症。为研究肝脏胰岛素抵抗而开发的体外模型有限，且所采用的细胞系与原代人肝细胞没有相似特征。Huh7细胞系已被确立为具有与原代人肝细胞相似特征的模型。此外，在Huh7细胞系中已发现丙型肝炎病毒感染会诱导胰岛素抵抗。因此，我们使用过量胰岛素诱导Huh7细胞系产生胰岛素抵抗（IR），并用二甲双胍进行治疗以逆转该抵抗，目的是建立一个对代谢和药理学研究有用的胰岛素抵抗模型。胰岛素抵抗的Huh7细胞（Huh7-IR）在胰岛素刺激后，其葡萄糖转运蛋白2（Glut2）、糖原水平以及胰岛素或胰岛素受体β亚基酪氨酸磷酸化刺激的葡萄糖摄取均降低，同时葡萄糖生成和细胞内脂质含量增加。这些生物标志物在胰岛素抵抗的肝细胞中经常被观察到。此外，用0.5、1或2 mM二甲双胍处理Huh7-IR 24小时可降低与胰岛素抵抗状态相关的生物标志物。这些结果表明，Huh7-IR可作为体外系统用于代谢和药理学研究中的肝脏胰岛素抵抗研究。
