Hyperglycemia induces key genetic and phenotypic changes in human liver epithelial HepG2 cells which parallel the Leprdb/J mouse model of non-alcoholic fatty liver disease (NAFLD)
Autoři:
Robin C. Su aff001; Apurva Lad aff001; Joshua D. Breidenbach aff001; Thomas M. Blomquist aff002; William T. Gunning aff002; Prabhatchandra Dube aff001; Andrew L. Kleinhenz aff001; Deepak Malhotra aff001; Steven T. Haller aff001; David J. Kennedy aff001
Působiště autorů:
Department of Medicine, The University of Toledo College of Medicine and Life Sciences, Toledo, Ohio, United States of America
aff001; Department of Pathology, The University of Toledo College of Medicine and Life Sciences, Toledo, Ohio, United States of America
aff002; Department of Medical Microbiology and Immunology, The University of Toledo College of Medicine and Life Sciences, Toledo, Ohio, United States of America
aff003
Vyšlo v časopise:
PLoS ONE 14(12)
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pone.0225604
Souhrn
Non-alcoholic fatty liver disease (NAFLD) is a growing global health concern. With a propensity to progress towards non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma, NAFLD is an important link amongst a multitude of comorbidities including obesity, diabetes, and cardiovascular and kidney disease. As several in vivo models of hyperglycemia and NAFLD are employed to investigate the pathophysiology of this disease process, we aimed to characterize an in vitro model of hyperglycemia that was amenable to address molecular mechanisms and therapeutic targets at the cellular level. Utilizing hyperglycemic cell culturing conditions, we induced steatosis within a human hepatocyte cell line (HepG2 cells), as confirmed by electron microscopy. The deposition and accumulation of lipids within hyperglycemic HepG2 cells is significantly greater than in normoglycemic cells, as visualized and quantified by Nile red staining. Alanine aminotransferase (ALT) and alkaline phosphatase (ALP), diagnostic biomarkers for liver damage and disease, were found to be upregulated in hyperglycemic HepG2 cells as compared with normoglycemic cells. Suppression of CEACAM1, GLUT2, and PON1, and elevation of CD36, PCK1, and G6PK were also found to be characteristic in hyperglycemic HepG2 cells compared with normoglycemic cells, suggesting insulin resistance and NAFLD. These in vitro findings mirror the characteristic genetic and phenotypic profile seen in Leprdb/J mice, a well-established in vivo model of NAFLD. In conclusion, we characterize an in vitro model displaying several key genetic and phenotypic characteristics in common with NAFLD that may assist future studies in addressing the molecular mechanisms and therapeutic targets to combat this disease.
Klíčová slova:
Gene expression – Lipids – Mouse models – Cell staining – MTT assay – Fluorescence imaging – Fatty liver
Zdroje
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