RESUMEN
Hormones regulate hepatic gene expressions to maintain metabolic homeostasis. Ectonucleotide pyrophosphatase/phosphodiesterase 1 has been thought to interfere with insulin signaling. To determine its potential role in the regulation of metabolism, we analyzed its gene (Enpp1) expression in the liver of rats experiencing fasting and refeeding cycles, and in primary rat hepatocytes and human hepatoma HepG2 cells treated with insulin and dexamethasone using northern blot and real-time PCR techniques. Hepatic Enpp1 expression was induced by fasting and reduced by refeeding in the rat liver. In primary rat hepatocytes and HepG2 hepatoma cells, insulin reduced Enpp1 mRNA abundance, whereas dexamethasone induced it. Dexamethasone disrupted the insulin-reduced Enpp1 expression in primary hepatocytes. This is in contrast to the responses of the expression of the cytosolic form of phosphoenolpyruvate carboxykinase gene to the same hormones, where insulin reduced it significantly in the process. In addition, the dexamethasone-induced Enpp1 gene expression was attenuated in the presence of 8-Br-cAMP. In conclusion, we demonstrated for the first time that hepatic Enpp1 is regulated in the cycle of fasting and refeeding, a process that might be attributed to insulin-reduced Enpp1 expression. This insulin-reduced Enpp1 expression might play a role in the development of complications in diabetic patients.
Asunto(s)
Humanos , Animales , Masculino , Ratas , Pirofosfatasas/genética , ARN Mensajero/efectos de los fármacos , Dexametasona/farmacología , Hidrolasas Diéster Fosfóricas/genética , Glucocorticoides/farmacología , Hipoglucemiantes/farmacología , Insulina/farmacología , Hígado/enzimología , Pirofosfatasas/biosíntesis , Pirofosfatasas/efectos de los fármacos , Resistencia a la Insulina , ARN Mensajero/metabolismo , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Inducción Enzimática/efectos de los fármacos , Ayuno/metabolismo , Ratas Sprague-Dawley , Hidrolasas Diéster Fosfóricas/biosíntesis , Hidrolasas Diéster Fosfóricas/efectos de los fármacos , Células Hep G2 , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
A mitochondrial pyrophosphatase (PPase) from yeast cells (Saccharomyces cerevisiae) was studied and characterized. The hydrolytic activity towards inorganic pyrophosphate (PPi) was inhibited by different SH-reagents and increased in the presence of uncouplers, indicating a possible involvement of this enzyme in energy-linked processes. This view was also supported by the observation that these mitochondria were able to hydrolyze PPi, generating an electrical membrane potential (delta psi) of the same magnitude as that obtained with ATP. Both ATP and PPi inhibited the pyruvate dehydrogenase complex and it was demonstrated that PPi can be used as substrate by mitochondrial kinases leading to the same pattern of protein phosphorylation as when ATP is used