Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesicles

The adipokine Neuregulin 4 (Nrg4) protects against obesity-induced insulin resistance. Here, we analyze how the downregulation of Nrg4 influences insulin action and the underlying mechanisms in adipocytes. Validated shRNA lentiviral vectors were used to generate scramble (Scr) and Nrg4 knockdown (KD...

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Autores: Díaz-Sáez, Francisco, Blanco-Sinfreu, Carla, Archilla-Ortega, Adrià, Sebastian, David, Romero, Montserrat, Hernández-Alvarez, Maria Isabel, Mora, Sílvia, Testar, Xavier, Ricart, Wifredo, Fernández-Real, José Manuel, Moreno-Navarrete, José María, Aragonés López, Julián, Camps, Marta, Zorzano, Antonio, Gumà, Anna
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Universidad Autónoma de Madrid
Repositorio:Biblos-e Archivo. Repositorio Institucional de la UAM
Idioma:inglés
OAI Identifier:oai:repositorio.uam.es:10486/716908
Acceso en línea:http://hdl.handle.net/10486/716908
https://dx.doi.org/10.3390/ijms222312960
Access Level:acceso abierto
Palabra clave:Adipocytes
Autophagy
Glut4
inflammation
insulin receptor
insulin resistance
Neuregulin 4
Medicina
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spelling Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesiclesDíaz-Sáez, FranciscoBlanco-Sinfreu, CarlaArchilla-Ortega, AdriàSebastian, DavidRomero, MontserratHernández-Alvarez, Maria IsabelMora, SílviaTestar, XavierRicart, WifredoFernández-Real, José ManuelMoreno-Navarrete, José MaríaAragonés López, JuliánCamps, MartaZorzano, AntonioGumà, AnnaAdipocytesAutophagyGlut4inflammationinsulin receptorinsulin resistanceNeuregulin 4MedicinaThe adipokine Neuregulin 4 (Nrg4) protects against obesity-induced insulin resistance. Here, we analyze how the downregulation of Nrg4 influences insulin action and the underlying mechanisms in adipocytes. Validated shRNA lentiviral vectors were used to generate scramble (Scr) and Nrg4 knockdown (KD) 3T3-L1 adipocytes. Adipogenesis was unaffected in Nrg4 KD adipocytes, but there was a complete impairment of the insulin-induced 2-deoxyglucose uptake, which was likely the result of reduced insulin receptor and Glut4 protein. Downregulation of Nrg4 enhanced the expression of proinflammatory cytokines. Anti-inflammatory agents recovered the insulin receptor, but not Glut4, content. Proteins enriched in Glut4 storage vesicles such as the insulin-responsive aminopeptidase (IRAP) and Syntaxin-6 as well as TBC1D4, a protein involved in the intracellular retention of Glut4 vesicles, also decreased by Nrg4 KD. Insulin failed to reduce autophagy in Nrg4 KD adipocytes, observed by a minor effect on mTOR phosphorylation, at the time that proteins involved in autophagy such as LC3-II, Rab11, and Clathrin were markedly upregulated. The lysosomal activity inhibitor bafilomycin A1 restored Glut4, IRAP, Syntaxin-6, and TBC1D4 content to those found in control adipocytes. Our study reveals that Nrg4 preserves the insulin responsiveness by preventing inflammation and, in turn, benefits the insulin regulation of autophagyGrant 534/C/2016 and 201626-30 funded by FUNDACIÓ LA MARATÓ DE TV3, Catalonia, Spain; Grants SAF2016-75246R, SAF2016-76815 and PID2019-105466RA-100 funded by MCIN/AEI/10.13039/501100011033, Madrid, Spain, by “ERDF A way of making Europe”, by the “European Union”; Grant 2017SGR1015 funded by GENERALITAT DE CATALUNYA, Catalonia, Spain; Grant PIE-14/00045 as well as Programs CIBERDEM and INFLAMES funded by ISCIII, Madrid, Spain;¸Grant CIVP18A3942 funded by FUNDACIÓN RAMÓN ARECES, Madrid, SpainMDPIDepartamento de MedicinaFacultad de Medicina20212021-12-01research articlehttp://purl.org/coar/resource_type/c_2df8fbb1VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10486/716908https://dx.doi.org/10.3390/ijms222312960reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridInglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/7169082026-06-23T12:46:27Z
dc.title.none.fl_str_mv Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesicles
title Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesicles
spellingShingle Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesicles
Díaz-Sáez, Francisco
Adipocytes
Autophagy
Glut4
inflammation
insulin receptor
insulin resistance
Neuregulin 4
Medicina
title_short Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesicles
title_full Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesicles
title_fullStr Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesicles
title_full_unstemmed Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesicles
title_sort Neuregulin 4 downregulation induces insulin resistance in 3t3-l1 adipocytes through inflammation and autophagic degradation of glut4 vesicles
dc.creator.none.fl_str_mv Díaz-Sáez, Francisco
Blanco-Sinfreu, Carla
Archilla-Ortega, Adrià
Sebastian, David
Romero, Montserrat
Hernández-Alvarez, Maria Isabel
Mora, Sílvia
Testar, Xavier
Ricart, Wifredo
Fernández-Real, José Manuel
Moreno-Navarrete, José María
Aragonés López, Julián
Camps, Marta
Zorzano, Antonio
Gumà, Anna
author Díaz-Sáez, Francisco
author_facet Díaz-Sáez, Francisco
Blanco-Sinfreu, Carla
Archilla-Ortega, Adrià
Sebastian, David
Romero, Montserrat
Hernández-Alvarez, Maria Isabel
Mora, Sílvia
Testar, Xavier
Ricart, Wifredo
Fernández-Real, José Manuel
Moreno-Navarrete, José María
Aragonés López, Julián
Camps, Marta
Zorzano, Antonio
Gumà, Anna
author_role author
author2 Blanco-Sinfreu, Carla
Archilla-Ortega, Adrià
Sebastian, David
Romero, Montserrat
Hernández-Alvarez, Maria Isabel
Mora, Sílvia
Testar, Xavier
Ricart, Wifredo
Fernández-Real, José Manuel
Moreno-Navarrete, José María
Aragonés López, Julián
Camps, Marta
Zorzano, Antonio
Gumà, Anna
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Departamento de Medicina
Facultad de Medicina
dc.subject.none.fl_str_mv Adipocytes
Autophagy
Glut4
inflammation
insulin receptor
insulin resistance
Neuregulin 4
Medicina
topic Adipocytes
Autophagy
Glut4
inflammation
insulin receptor
insulin resistance
Neuregulin 4
Medicina
description The adipokine Neuregulin 4 (Nrg4) protects against obesity-induced insulin resistance. Here, we analyze how the downregulation of Nrg4 influences insulin action and the underlying mechanisms in adipocytes. Validated shRNA lentiviral vectors were used to generate scramble (Scr) and Nrg4 knockdown (KD) 3T3-L1 adipocytes. Adipogenesis was unaffected in Nrg4 KD adipocytes, but there was a complete impairment of the insulin-induced 2-deoxyglucose uptake, which was likely the result of reduced insulin receptor and Glut4 protein. Downregulation of Nrg4 enhanced the expression of proinflammatory cytokines. Anti-inflammatory agents recovered the insulin receptor, but not Glut4, content. Proteins enriched in Glut4 storage vesicles such as the insulin-responsive aminopeptidase (IRAP) and Syntaxin-6 as well as TBC1D4, a protein involved in the intracellular retention of Glut4 vesicles, also decreased by Nrg4 KD. Insulin failed to reduce autophagy in Nrg4 KD adipocytes, observed by a minor effect on mTOR phosphorylation, at the time that proteins involved in autophagy such as LC3-II, Rab11, and Clathrin were markedly upregulated. The lysosomal activity inhibitor bafilomycin A1 restored Glut4, IRAP, Syntaxin-6, and TBC1D4 content to those found in control adipocytes. Our study reveals that Nrg4 preserves the insulin responsiveness by preventing inflammation and, in turn, benefits the insulin regulation of autophagy
publishDate 2021
dc.date.none.fl_str_mv 2021
2021-12-01
dc.type.none.fl_str_mv research article
http://purl.org/coar/resource_type/c_2df8fbb1
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10486/716908
https://dx.doi.org/10.3390/ijms222312960
url http://hdl.handle.net/10486/716908
https://dx.doi.org/10.3390/ijms222312960
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Biblos-e Archivo. Repositorio Institucional de la UAM
instname:Universidad Autónoma de Madrid
instname_str Universidad Autónoma de Madrid
reponame_str Biblos-e Archivo. Repositorio Institucional de la UAM
collection Biblos-e Archivo. Repositorio Institucional de la UAM
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