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Increased Phagocyte-Like NADPH Oxidase and ROS Generation in Type 2 Diabetic ZDF Rat and Human Islets: Role of Rac1–JNK1/2 Signaling Pathway in Mitochondrial Dysregulation in the Diabetic Islet
OBJECTIVE: To determine the subunit expression and functional activation of phagocyte-like NADPH oxidase (Nox), reactive oxygen species (ROS) generation and caspase-3 activation in the Zucker diabetic fatty (ZDF) rat and diabetic human islets. RESEARCH DESIGN AND METHODS: Expression of core componen...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Diabetes Association
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3198065/ https://www.ncbi.nlm.nih.gov/pubmed/21911753 http://dx.doi.org/10.2337/db11-0809 |
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author | Syed, Ismail Kyathanahalli, Chandrashekara N. Jayaram, Bhavaani Govind, Sudha Rhodes, Christopher J. Kowluru, Renu A. Kowluru, Anjaneyulu |
author_facet | Syed, Ismail Kyathanahalli, Chandrashekara N. Jayaram, Bhavaani Govind, Sudha Rhodes, Christopher J. Kowluru, Renu A. Kowluru, Anjaneyulu |
author_sort | Syed, Ismail |
collection | PubMed |
description | OBJECTIVE: To determine the subunit expression and functional activation of phagocyte-like NADPH oxidase (Nox), reactive oxygen species (ROS) generation and caspase-3 activation in the Zucker diabetic fatty (ZDF) rat and diabetic human islets. RESEARCH DESIGN AND METHODS: Expression of core components of Nox was quantitated by Western blotting and densitometry. ROS levels were quantitated by the 2′,7′-dichlorofluorescein diacetate method. Rac1 activation was quantitated using the gold-labeled immunosorbent assay kit. RESULTS: Levels of phosphorylated p47(phox), active Rac1, Nox activity, ROS generation, Jun NH(2)-terminal kinase (JNK) 1/2 phosphorylation, and caspase-3 activity were significantly higher in the ZDF islets than the lean control rat islets. Chronic exposure of INS 832/13 cells to glucolipotoxic conditions resulted in increased JNK1/2 phosphorylation and caspase-3 activity; such effects were largely reversed by SP600125, a selective inhibitor of JNK. Incubation of normal human islets with high glucose also increased the activation of Rac1 and Nox. Lastly, in a manner akin to the ZDF diabetic rat islets, Rac1 expression, JNK1/2, and caspase-3 activation were also significantly increased in diabetic human islets. CONCLUSIONS: We provide the first in vitro and in vivo evidence in support of an accelerated Rac1–Nox–ROS–JNK1/2 signaling pathway in the islet β-cell leading to the onset of mitochondrial dysregulation in diabetes. |
format | Online Article Text |
id | pubmed-3198065 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | American Diabetes Association |
record_format | MEDLINE/PubMed |
spelling | pubmed-31980652012-11-01 Increased Phagocyte-Like NADPH Oxidase and ROS Generation in Type 2 Diabetic ZDF Rat and Human Islets: Role of Rac1–JNK1/2 Signaling Pathway in Mitochondrial Dysregulation in the Diabetic Islet Syed, Ismail Kyathanahalli, Chandrashekara N. Jayaram, Bhavaani Govind, Sudha Rhodes, Christopher J. Kowluru, Renu A. Kowluru, Anjaneyulu Diabetes Islet Studies OBJECTIVE: To determine the subunit expression and functional activation of phagocyte-like NADPH oxidase (Nox), reactive oxygen species (ROS) generation and caspase-3 activation in the Zucker diabetic fatty (ZDF) rat and diabetic human islets. RESEARCH DESIGN AND METHODS: Expression of core components of Nox was quantitated by Western blotting and densitometry. ROS levels were quantitated by the 2′,7′-dichlorofluorescein diacetate method. Rac1 activation was quantitated using the gold-labeled immunosorbent assay kit. RESULTS: Levels of phosphorylated p47(phox), active Rac1, Nox activity, ROS generation, Jun NH(2)-terminal kinase (JNK) 1/2 phosphorylation, and caspase-3 activity were significantly higher in the ZDF islets than the lean control rat islets. Chronic exposure of INS 832/13 cells to glucolipotoxic conditions resulted in increased JNK1/2 phosphorylation and caspase-3 activity; such effects were largely reversed by SP600125, a selective inhibitor of JNK. Incubation of normal human islets with high glucose also increased the activation of Rac1 and Nox. Lastly, in a manner akin to the ZDF diabetic rat islets, Rac1 expression, JNK1/2, and caspase-3 activation were also significantly increased in diabetic human islets. CONCLUSIONS: We provide the first in vitro and in vivo evidence in support of an accelerated Rac1–Nox–ROS–JNK1/2 signaling pathway in the islet β-cell leading to the onset of mitochondrial dysregulation in diabetes. American Diabetes Association 2011-11 2011-10-17 /pmc/articles/PMC3198065/ /pubmed/21911753 http://dx.doi.org/10.2337/db11-0809 Text en © 2011 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details. |
spellingShingle | Islet Studies Syed, Ismail Kyathanahalli, Chandrashekara N. Jayaram, Bhavaani Govind, Sudha Rhodes, Christopher J. Kowluru, Renu A. Kowluru, Anjaneyulu Increased Phagocyte-Like NADPH Oxidase and ROS Generation in Type 2 Diabetic ZDF Rat and Human Islets: Role of Rac1–JNK1/2 Signaling Pathway in Mitochondrial Dysregulation in the Diabetic Islet |
title | Increased Phagocyte-Like NADPH Oxidase and ROS Generation in Type 2 Diabetic ZDF Rat and Human Islets: Role of Rac1–JNK1/2 Signaling Pathway in Mitochondrial Dysregulation in the Diabetic Islet |
title_full | Increased Phagocyte-Like NADPH Oxidase and ROS Generation in Type 2 Diabetic ZDF Rat and Human Islets: Role of Rac1–JNK1/2 Signaling Pathway in Mitochondrial Dysregulation in the Diabetic Islet |
title_fullStr | Increased Phagocyte-Like NADPH Oxidase and ROS Generation in Type 2 Diabetic ZDF Rat and Human Islets: Role of Rac1–JNK1/2 Signaling Pathway in Mitochondrial Dysregulation in the Diabetic Islet |
title_full_unstemmed | Increased Phagocyte-Like NADPH Oxidase and ROS Generation in Type 2 Diabetic ZDF Rat and Human Islets: Role of Rac1–JNK1/2 Signaling Pathway in Mitochondrial Dysregulation in the Diabetic Islet |
title_short | Increased Phagocyte-Like NADPH Oxidase and ROS Generation in Type 2 Diabetic ZDF Rat and Human Islets: Role of Rac1–JNK1/2 Signaling Pathway in Mitochondrial Dysregulation in the Diabetic Islet |
title_sort | increased phagocyte-like nadph oxidase and ros generation in type 2 diabetic zdf rat and human islets: role of rac1–jnk1/2 signaling pathway in mitochondrial dysregulation in the diabetic islet |
topic | Islet Studies |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3198065/ https://www.ncbi.nlm.nih.gov/pubmed/21911753 http://dx.doi.org/10.2337/db11-0809 |
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