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SUN-651 Murine Cecal Ligation and Puncture (CLP) Perturbs Phosphorylation of Insulin Receptor Substrate 2 (IRS-2)

Hyperglycemia is a characteristic finding in sepsis, and its presence worsens outcome (1). Patients with sepsis need larger doses of insulin to reduce glucose levels. This abnormality has been termed “insulin resistance” but the molecular mechanism by which sepsis attenuates the insulin signaling pa...

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Autores principales: Mathew, Deepa, Barillas, Julia, Fernandes, Tiago, Kelly, Alexander, Yaipen, Omar, Abraham, Mabel, Deutschman, Clifford
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7208800/
http://dx.doi.org/10.1210/jendso/bvaa046.650
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author Mathew, Deepa
Barillas, Julia
Fernandes, Tiago
Kelly, Alexander
Yaipen, Omar
Abraham, Mabel
Deutschman, Clifford
author_facet Mathew, Deepa
Barillas, Julia
Fernandes, Tiago
Kelly, Alexander
Yaipen, Omar
Abraham, Mabel
Deutschman, Clifford
author_sort Mathew, Deepa
collection PubMed
description Hyperglycemia is a characteristic finding in sepsis, and its presence worsens outcome (1). Patients with sepsis need larger doses of insulin to reduce glucose levels. This abnormality has been termed “insulin resistance” but the molecular mechanism by which sepsis attenuates the insulin signaling pathway is unknown. Previous work has shown impairment of phosphorylation in several intracellular signaling pathways following CLP, a well-validated murine model of sepsis (2). Phosphorylation of tyrosine in IRS-2 is essential for functional insulin signaling in hepatocytes (3). Therefore the aim of this study was to investigate the effects of CLP on IRS-2 phosphorylation. Hypothesis: CLP attenuates phosphorylation of IRS-2. Methods: All studies were approved by the Feinstein IACUC and conformed to ARRIVE guidelines. CLP was performed on C57Bl6 mice. Before CLP, animals were identified for sacrifice at specific post-procedure time points. To stimulate phosphorylation of IRS-2, insulin was injected in control and CLP mice at 24 and 48 hours post CLP. Following sacrifice, protein was isolated from liver tissue. Protein abundance was determined using immunoblotting. The detection of the phosphorylated form of these proteins was determined by enzyme-linked immunosorbent assay (ELISA) with a phospho-insulin receptor antibody. Statistical significance was determined using ANOVA for repeated measures with a Sidak post-hoc correction. Results: Relative to the control, tyrosine phosphorylation of IRS-2 was significantly (p<0.05) reduced at 24 and 48 hours following CLP. Conclusions: Tyrosine phosphorylation of hepatic IRS2 is attenuated at early time points following CLP. These results are consistent with other studies examining the effects of CLP on intra-cellular signal transduction pathways (1). Further, these results provide evidence that changes in the insulin signaling transduction underlie CLP-induced “insulin resistance”. References: (1) Abcejo et al., Crit Care Med. 2009;37(5):1729-1734. (2) van den Berghe et al, NEngl J Med. 2001;345(19):1359-1367. (3) Valverde et al., Diabetes 2003 Sep; 52(9): 2239-2248.
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spelling pubmed-72088002020-05-13 SUN-651 Murine Cecal Ligation and Puncture (CLP) Perturbs Phosphorylation of Insulin Receptor Substrate 2 (IRS-2) Mathew, Deepa Barillas, Julia Fernandes, Tiago Kelly, Alexander Yaipen, Omar Abraham, Mabel Deutschman, Clifford J Endocr Soc Diabetes Mellitus and Glucose Metabolism Hyperglycemia is a characteristic finding in sepsis, and its presence worsens outcome (1). Patients with sepsis need larger doses of insulin to reduce glucose levels. This abnormality has been termed “insulin resistance” but the molecular mechanism by which sepsis attenuates the insulin signaling pathway is unknown. Previous work has shown impairment of phosphorylation in several intracellular signaling pathways following CLP, a well-validated murine model of sepsis (2). Phosphorylation of tyrosine in IRS-2 is essential for functional insulin signaling in hepatocytes (3). Therefore the aim of this study was to investigate the effects of CLP on IRS-2 phosphorylation. Hypothesis: CLP attenuates phosphorylation of IRS-2. Methods: All studies were approved by the Feinstein IACUC and conformed to ARRIVE guidelines. CLP was performed on C57Bl6 mice. Before CLP, animals were identified for sacrifice at specific post-procedure time points. To stimulate phosphorylation of IRS-2, insulin was injected in control and CLP mice at 24 and 48 hours post CLP. Following sacrifice, protein was isolated from liver tissue. Protein abundance was determined using immunoblotting. The detection of the phosphorylated form of these proteins was determined by enzyme-linked immunosorbent assay (ELISA) with a phospho-insulin receptor antibody. Statistical significance was determined using ANOVA for repeated measures with a Sidak post-hoc correction. Results: Relative to the control, tyrosine phosphorylation of IRS-2 was significantly (p<0.05) reduced at 24 and 48 hours following CLP. Conclusions: Tyrosine phosphorylation of hepatic IRS2 is attenuated at early time points following CLP. These results are consistent with other studies examining the effects of CLP on intra-cellular signal transduction pathways (1). Further, these results provide evidence that changes in the insulin signaling transduction underlie CLP-induced “insulin resistance”. References: (1) Abcejo et al., Crit Care Med. 2009;37(5):1729-1734. (2) van den Berghe et al, NEngl J Med. 2001;345(19):1359-1367. (3) Valverde et al., Diabetes 2003 Sep; 52(9): 2239-2248. Oxford University Press 2020-05-08 /pmc/articles/PMC7208800/ http://dx.doi.org/10.1210/jendso/bvaa046.650 Text en © Endocrine Society 2020. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Diabetes Mellitus and Glucose Metabolism
Mathew, Deepa
Barillas, Julia
Fernandes, Tiago
Kelly, Alexander
Yaipen, Omar
Abraham, Mabel
Deutschman, Clifford
SUN-651 Murine Cecal Ligation and Puncture (CLP) Perturbs Phosphorylation of Insulin Receptor Substrate 2 (IRS-2)
title SUN-651 Murine Cecal Ligation and Puncture (CLP) Perturbs Phosphorylation of Insulin Receptor Substrate 2 (IRS-2)
title_full SUN-651 Murine Cecal Ligation and Puncture (CLP) Perturbs Phosphorylation of Insulin Receptor Substrate 2 (IRS-2)
title_fullStr SUN-651 Murine Cecal Ligation and Puncture (CLP) Perturbs Phosphorylation of Insulin Receptor Substrate 2 (IRS-2)
title_full_unstemmed SUN-651 Murine Cecal Ligation and Puncture (CLP) Perturbs Phosphorylation of Insulin Receptor Substrate 2 (IRS-2)
title_short SUN-651 Murine Cecal Ligation and Puncture (CLP) Perturbs Phosphorylation of Insulin Receptor Substrate 2 (IRS-2)
title_sort sun-651 murine cecal ligation and puncture (clp) perturbs phosphorylation of insulin receptor substrate 2 (irs-2)
topic Diabetes Mellitus and Glucose Metabolism
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7208800/
http://dx.doi.org/10.1210/jendso/bvaa046.650
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