Contents lists available at ScienceDirect Growth Hormone & IGF Research journal homepage: www.elsevier.com/locate/ghir Growth hormone ameliorates high glucose-induced steatosis on in vitro cultured human HepG2 hepatocytes by inhibiting de novo lipogenesis via ChREBP and FAS suppression Eréndira Villanueva-Ortega a,b , Lucia A. Méndez-García a , Guadalupe N. Garibay-Nieto b , Estibalitz Laresgoiti-Servitje c , Patricia Medina-Bravo d , Alfonso Olivos-García e , Martín H. Muñoz-Ortega f , Javier Ventura-Juárez f , Galileo Escobedo a, ⁎ a Laboratory for Proteomics and Metabolomics, Research Division, General Hospital of Mexico “Dr. Eduardo Liceaga”, 06720, Mexico City, Mexico. b Department of Genetics, General Hospital of Mexico “Dr. Eduardo Liceaga”, 06720, Mexico City, Mexico. c Clinical Medical Sciences, School of Medicine, Tecnológico de Monterrey, Campus Ciudad de México, 14380, Mexico City, Mexico. d Endocrinology Department, Hospital Infantil de México Federico Gómez, 06720, Mexico City, Mexico. e Experimental Research Unit, School of Medicine, Universidad Nacional Autónoma de México, General Hospital of Mexico “Dr. Eduardo Liceaga”, 06720, Mexico City, Mexico. f Universidad Autónoma de Aguascalientes, Departamento de Morfología, Centro de Ciencias Básicas, Edificio 202, Av. Universidad 940 Ciudad Universitaria C.P. 20130, Aguascalientes, Ags., Mexico. ARTICLE INFO Keywords: Growth hormone Steatosis IGF-1 ChREBP FAS HepG2 cells ABSTRACT Objective: Growth hormone (GH) deficiency has been associated with increased steatosis but the molecular mechanism has not been fully elucidated. We investigated the effect of GH on lipid accumulation of HepG2 cells cultured on an in vitro steatosis model and examined the potential involvement of insulin-like growth factor 1 (IGF-1) as well as lipogenic and lipolytic molecules. Methods: Control and steatosis conditions were induced by culturing HepG2 cells with 5.5 or 25 mmol/l glucose for 24 h, respectively. Afterward, cells were exposed to 0, 5, 10 or 20 ng/ml GH for another 24 h. Lipid content was quantified as well as mRNA and protein levels of IGF-1, carbohydrate responsive element-binding protein (ChREBP), sterol regulatory element-binding protein 1c (SREBP1c), fatty acid synthase (FAS), carnitine palmitoyltransferase 1A (CPT1A), and peroxisome proliferator-activated receptor alpha (PPAR-alpha) by qPCR and western blot, respectively. Data were analyzed by one-way ANOVA and the Games-Howell post-hoc test. Results: In the steatosis model, HepG2 hepatocytes showed a significant 2-fold increase in lipid amount as compared to control cells. IGF-1 mRNA and protein levels were significantly increased in control cells exposed to 10 ng/ml GH, whereas high glucose abolished this effect. High glucose also significantly increased both mRNA and protein of ChREBP and FAS without having effect on SREBP1c, CPT1A and PPAR-alpha. However, GH inhibited ChREBP and FAS production, even in HepG2 hepatocytes cultured under steatosis conditions. Conclusions: Growth hormone ameliorates high glucose-induced steatosis in HepG2 cells by suppressing de novo lipogenesis via ChREBP and FAS down-regulation. Introduction Non-alcoholic fatty liver disease (NAFLD) is characterized by he- patic fat accumulation and encompasses a wide spectrum of liver dis- orders progressively ranging from simple steatosis to steatohepatitis, liver fibrosis, cirrhosis, and hepatocellular carcinoma (HCC) [1]. NAFLD is now considered the most common liver disease worldwide, and due to increasing rates of obesity, NAFLD might be soon the most frequent indication for liver transplantation in west countries [2]. For this reason, it is of great importance to understand the main mechan- isms leading to hepatic fat accumulation, with special emphasis on simple steatosis as the first stage of NAFLD development and progres- sion. Growth hormone (GH), also referred to as somatotropin, is a 191- https://doi.org/10.1016/j.ghir.2020.101332 Received 17 October 2019; Received in revised form 13 May 2020; Accepted 1 June 2020 ⁎ Corresponding author at: Laboratory for Proteomics and Metabolomics, Research Division, General Hospital of Mexico “Dr. Eduardo Liceaga”, 06726, Mexico City, Mexico. E-mail address: gescobedo@unam.mx (G. Escobedo). Growth Hormone & IGF Research 53–54 (2020) 101332 Available online 15 July 2020 1096-6374/ © 2020 Elsevier Ltd. All rights reserved. T