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Exposure to Perfluoro-Octanoic Acid Associated With Upstream Uncoupling of the Insulin Signaling in Human Hepatocyte Cell Line

Perfluoro–alkyl substances (PFAS) are chemical pollutants with prevalent stability and environmental persistence. Exposure to PFAS, particularly perfluoro-octanoic acid (PFOA), has been associated with increased diabetes-related cardiovascular mortality in subjects residing areas of high environment...

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Autores principales: De Toni, Luca, Di Nisio, Andrea, Rocca, Maria Santa, Guidolin, Diego, Della Marina, Alice, Bertazza, Loris, Sut, Stefania, Purpura, Edoardo, Pannella, Micaela, Garolla, Andrea, Foresta, Carlo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8446511/
https://www.ncbi.nlm.nih.gov/pubmed/34539566
http://dx.doi.org/10.3389/fendo.2021.632927
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author De Toni, Luca
Di Nisio, Andrea
Rocca, Maria Santa
Guidolin, Diego
Della Marina, Alice
Bertazza, Loris
Sut, Stefania
Purpura, Edoardo
Pannella, Micaela
Garolla, Andrea
Foresta, Carlo
author_facet De Toni, Luca
Di Nisio, Andrea
Rocca, Maria Santa
Guidolin, Diego
Della Marina, Alice
Bertazza, Loris
Sut, Stefania
Purpura, Edoardo
Pannella, Micaela
Garolla, Andrea
Foresta, Carlo
author_sort De Toni, Luca
collection PubMed
description Perfluoro–alkyl substances (PFAS) are chemical pollutants with prevalent stability and environmental persistence. Exposure to PFAS, particularly perfluoro-octanoic acid (PFOA), has been associated with increased diabetes-related cardiovascular mortality in subjects residing areas of high environmental contamination, however the exact pathogenic mechanism remains elusive. Here we used HepG2 cells, an in vitro model of human hepatocyte, to investigate the possible role of PFOA exposure in the alteration of hepatic glucose metabolism. HepG2 cells were exposed for 24 hours to PFOA at increasing concentration from 0 to 1000 ng/mL and then stimulated with 100 nm Insulin (Ins). The consequent effect on glycogen synthesis, glucose uptake and Glut-4 glucose transporter translocation was then evaluated by, respectively, Periodic Acid Schiff (PAS) staining, 2-deoxyglucose (2-DG) uptake assay and immunofluorescence. Exposure to PFOA was associated with reduced glycogen synthesis and glucose uptake, at concentration equal or greater than, respectively, 0,1 ng/mL and 10 ng/mL, with parallel impaired membrane translocation of Glut-4 upon Ins stimulation. Western blot analysis showed early uncoupling of Insulin Receptor (InsR) activation from the downstream Akt and GSK3 phosphorylation. Computational docking analysis disclosed the possible stabilizing effect of PFOA on the complex between InsR and GM3 ganglioside, previously shown to be associated with the low grade chronic inflammation-related insulin resistance. Consistently, long term treatment with glucosyl-ceramide synthase inhibitor PDMP was able to largely restore glycogen synthesis, glucose uptake and Glut-4 translocation upon Ins stimulation in HepG2 exposed to PFOA. Our data support a novel pathogenic mechanism linking exposure to PFOA to derangement of hepatocyte cell metabolism.
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spelling pubmed-84465112021-09-18 Exposure to Perfluoro-Octanoic Acid Associated With Upstream Uncoupling of the Insulin Signaling in Human Hepatocyte Cell Line De Toni, Luca Di Nisio, Andrea Rocca, Maria Santa Guidolin, Diego Della Marina, Alice Bertazza, Loris Sut, Stefania Purpura, Edoardo Pannella, Micaela Garolla, Andrea Foresta, Carlo Front Endocrinol (Lausanne) Endocrinology Perfluoro–alkyl substances (PFAS) are chemical pollutants with prevalent stability and environmental persistence. Exposure to PFAS, particularly perfluoro-octanoic acid (PFOA), has been associated with increased diabetes-related cardiovascular mortality in subjects residing areas of high environmental contamination, however the exact pathogenic mechanism remains elusive. Here we used HepG2 cells, an in vitro model of human hepatocyte, to investigate the possible role of PFOA exposure in the alteration of hepatic glucose metabolism. HepG2 cells were exposed for 24 hours to PFOA at increasing concentration from 0 to 1000 ng/mL and then stimulated with 100 nm Insulin (Ins). The consequent effect on glycogen synthesis, glucose uptake and Glut-4 glucose transporter translocation was then evaluated by, respectively, Periodic Acid Schiff (PAS) staining, 2-deoxyglucose (2-DG) uptake assay and immunofluorescence. Exposure to PFOA was associated with reduced glycogen synthesis and glucose uptake, at concentration equal or greater than, respectively, 0,1 ng/mL and 10 ng/mL, with parallel impaired membrane translocation of Glut-4 upon Ins stimulation. Western blot analysis showed early uncoupling of Insulin Receptor (InsR) activation from the downstream Akt and GSK3 phosphorylation. Computational docking analysis disclosed the possible stabilizing effect of PFOA on the complex between InsR and GM3 ganglioside, previously shown to be associated with the low grade chronic inflammation-related insulin resistance. Consistently, long term treatment with glucosyl-ceramide synthase inhibitor PDMP was able to largely restore glycogen synthesis, glucose uptake and Glut-4 translocation upon Ins stimulation in HepG2 exposed to PFOA. Our data support a novel pathogenic mechanism linking exposure to PFOA to derangement of hepatocyte cell metabolism. Frontiers Media S.A. 2021-09-03 /pmc/articles/PMC8446511/ /pubmed/34539566 http://dx.doi.org/10.3389/fendo.2021.632927 Text en Copyright © 2021 De Toni, Di Nisio, Rocca, Guidolin, Della Marina, Bertazza, Sut, Purpura, Pannella, Garolla and Foresta https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Endocrinology
De Toni, Luca
Di Nisio, Andrea
Rocca, Maria Santa
Guidolin, Diego
Della Marina, Alice
Bertazza, Loris
Sut, Stefania
Purpura, Edoardo
Pannella, Micaela
Garolla, Andrea
Foresta, Carlo
Exposure to Perfluoro-Octanoic Acid Associated With Upstream Uncoupling of the Insulin Signaling in Human Hepatocyte Cell Line
title Exposure to Perfluoro-Octanoic Acid Associated With Upstream Uncoupling of the Insulin Signaling in Human Hepatocyte Cell Line
title_full Exposure to Perfluoro-Octanoic Acid Associated With Upstream Uncoupling of the Insulin Signaling in Human Hepatocyte Cell Line
title_fullStr Exposure to Perfluoro-Octanoic Acid Associated With Upstream Uncoupling of the Insulin Signaling in Human Hepatocyte Cell Line
title_full_unstemmed Exposure to Perfluoro-Octanoic Acid Associated With Upstream Uncoupling of the Insulin Signaling in Human Hepatocyte Cell Line
title_short Exposure to Perfluoro-Octanoic Acid Associated With Upstream Uncoupling of the Insulin Signaling in Human Hepatocyte Cell Line
title_sort exposure to perfluoro-octanoic acid associated with upstream uncoupling of the insulin signaling in human hepatocyte cell line
topic Endocrinology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8446511/
https://www.ncbi.nlm.nih.gov/pubmed/34539566
http://dx.doi.org/10.3389/fendo.2021.632927
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