Establishment and characterization of a novel method for evaluating gluconeogenesis using hepatic cell lines, H4IIE and HepG2

The liver gluconeogenic pathway is recognized as a target for treating diabetes mellitus. In this study, we attempted to establish a new method to evaluate gluconeogenesis using rat H4IIE hepatoma cells. High-density preculture and exposure to hypertonic solutions, which are known to upregulate the...

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Bibliographic Details
Published in:Archives of biochemistry and biophysics 2009-11, Vol.491 (1), p.46-52
Main Authors: Okamoto, Takashi, Kanemoto, Naohide, Ban, Takashi, Sudo, Toshiki, Nagano, Keisuke, Niki, Ichiro
Format: Article
Language:English
Subjects:
Animals
Cell Culture Techniques
Cell Line, Tumor
Fluorometry
Gluconeogenesis
Gluconeogenesis - drug effects
Glucose - biosynthesis
Glucose - metabolism
Glucose-6-phosphatase
Hepatocytes - metabolism
Hepatoma
Humans
Hypoglycemic Agents - pharmacology
Insulin - pharmacology
Lactic Acid - metabolism
Metformin
Metformin - pharmacology
Phosphoenolpyruvate carboxykinase
Pyruvic Acid - metabolism
Rats
Up-Regulation
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Summary:The liver gluconeogenic pathway is recognized as a target for treating diabetes mellitus. In this study, we attempted to establish a new method to evaluate gluconeogenesis using rat H4IIE hepatoma cells. High-density preculture and exposure to hypertonic solutions, which are known to upregulate the expression of gluconeogenic genes, enhanced glucose release (GR) promoted by gluconeogenic substrates (GS: 1 mM pyruvate and 10 mM lactate). Our method was also applicable to the human hepatoma HepG2 cells. Measurement of glycogen content in HepG2 cells revealed that GR was compensated by glycogenolysis in the basal state and was generated by gluconeogenesis in the presence of GS. The optimized conditions increased the expression of gluconeogenic genes in HepG2 cells. Insulin and metformin dose-dependently inhibited GR and 8-(4-chlorophenylthio)-cAMP (CPT-cAMP) increased it. These results suggest that the present method is useful to evaluate the effects of nutrients, hormones and hypoglycemic agents on hepatic gluconeogenesis.
ISSN:0003-9861
1096-0384
DOI:10.1016/j.abb.2009.09.015