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The Impaired Bioenergetics of Diabetic Cardiac Microvascular Endothelial Cells

Diabetes causes hyperglycemia, which can create a stressful environment for cardiac microvascular endothelial cells (CMECs). To investigate the impact of diabetes on the cellular metabolism of CMECs, we assessed glycolysis by quantifying the extracellular acidification rate (ECAR), and mitochondrial...

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Autores principales: Zhang, Haitao, Shen, Yan, Kim, Il-man, Weintraub, Neal L., Tang, Yaoliang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8160466/
https://www.ncbi.nlm.nih.gov/pubmed/34054724
http://dx.doi.org/10.3389/fendo.2021.642857
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author Zhang, Haitao
Shen, Yan
Kim, Il-man
Weintraub, Neal L.
Tang, Yaoliang
author_facet Zhang, Haitao
Shen, Yan
Kim, Il-man
Weintraub, Neal L.
Tang, Yaoliang
author_sort Zhang, Haitao
collection PubMed
description Diabetes causes hyperglycemia, which can create a stressful environment for cardiac microvascular endothelial cells (CMECs). To investigate the impact of diabetes on the cellular metabolism of CMECs, we assessed glycolysis by quantifying the extracellular acidification rate (ECAR), and mitochondrial oxidative phosphorylation (OXPHOS) by measuring cellular oxygen consumption rate (OCR), in isolated CMECs from wild-type (WT) hearts and diabetic hearts (db/db) using an extracellular flux analyzer. Diabetic CMECs exhibited a higher level of intracellular reactive oxygen species (ROS), and significantly reduced glycolytic reserve and non-glycolytic acidification, as compared to WT CMECs. In addition, OCR assay showed that diabetic CMECs had increased maximal respiration, and significantly reduced non-mitochondrial oxygen consumption and proton leak. Quantitative PCR (qPCR) showed no difference in copy number of mitochondrial DNA (mtDNA) between diabetic and WT CMECs. In addition, gene expression profiling analysis showed an overall decrease in the expression of essential genes related to β-oxidation (Sirt1, Acox1, Acox3, Hadha, and Hadhb), tricarboxylic acid cycle (TCA) (Idh-3a and Ogdh), and electron transport chain (ETC) (Sdhd and Uqcrq) in diabetic CMECs compared to WT CMECs. Western blot confirmed that the protein expression of Hadha, Acox1, and Uqcrq was decreased in diabetic CMECs. Although lectin staining demonstrated no significant difference in capillary density between the hearts of WT mice and db/db mice, diabetic CMECs showed a lower percentage of cell proliferation by Ki67 staining, and a higher percentage of cellular apoptosis by TUNEL staining, compared with WT CMECs. In conclusion, excessive ROS caused by hyperglycemia is associated with impaired glycolysis and mitochondrial function in diabetic CMECs, which in turn may reduce proliferation and promote CMEC apoptosis.
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spelling pubmed-81604662021-05-29 The Impaired Bioenergetics of Diabetic Cardiac Microvascular Endothelial Cells Zhang, Haitao Shen, Yan Kim, Il-man Weintraub, Neal L. Tang, Yaoliang Front Endocrinol (Lausanne) Endocrinology Diabetes causes hyperglycemia, which can create a stressful environment for cardiac microvascular endothelial cells (CMECs). To investigate the impact of diabetes on the cellular metabolism of CMECs, we assessed glycolysis by quantifying the extracellular acidification rate (ECAR), and mitochondrial oxidative phosphorylation (OXPHOS) by measuring cellular oxygen consumption rate (OCR), in isolated CMECs from wild-type (WT) hearts and diabetic hearts (db/db) using an extracellular flux analyzer. Diabetic CMECs exhibited a higher level of intracellular reactive oxygen species (ROS), and significantly reduced glycolytic reserve and non-glycolytic acidification, as compared to WT CMECs. In addition, OCR assay showed that diabetic CMECs had increased maximal respiration, and significantly reduced non-mitochondrial oxygen consumption and proton leak. Quantitative PCR (qPCR) showed no difference in copy number of mitochondrial DNA (mtDNA) between diabetic and WT CMECs. In addition, gene expression profiling analysis showed an overall decrease in the expression of essential genes related to β-oxidation (Sirt1, Acox1, Acox3, Hadha, and Hadhb), tricarboxylic acid cycle (TCA) (Idh-3a and Ogdh), and electron transport chain (ETC) (Sdhd and Uqcrq) in diabetic CMECs compared to WT CMECs. Western blot confirmed that the protein expression of Hadha, Acox1, and Uqcrq was decreased in diabetic CMECs. Although lectin staining demonstrated no significant difference in capillary density between the hearts of WT mice and db/db mice, diabetic CMECs showed a lower percentage of cell proliferation by Ki67 staining, and a higher percentage of cellular apoptosis by TUNEL staining, compared with WT CMECs. In conclusion, excessive ROS caused by hyperglycemia is associated with impaired glycolysis and mitochondrial function in diabetic CMECs, which in turn may reduce proliferation and promote CMEC apoptosis. Frontiers Media S.A. 2021-05-14 /pmc/articles/PMC8160466/ /pubmed/34054724 http://dx.doi.org/10.3389/fendo.2021.642857 Text en Copyright © 2021 Zhang, Shen, Kim, Weintraub and Tang 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
Zhang, Haitao
Shen, Yan
Kim, Il-man
Weintraub, Neal L.
Tang, Yaoliang
The Impaired Bioenergetics of Diabetic Cardiac Microvascular Endothelial Cells
title The Impaired Bioenergetics of Diabetic Cardiac Microvascular Endothelial Cells
title_full The Impaired Bioenergetics of Diabetic Cardiac Microvascular Endothelial Cells
title_fullStr The Impaired Bioenergetics of Diabetic Cardiac Microvascular Endothelial Cells
title_full_unstemmed The Impaired Bioenergetics of Diabetic Cardiac Microvascular Endothelial Cells
title_short The Impaired Bioenergetics of Diabetic Cardiac Microvascular Endothelial Cells
title_sort impaired bioenergetics of diabetic cardiac microvascular endothelial cells
topic Endocrinology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8160466/
https://www.ncbi.nlm.nih.gov/pubmed/34054724
http://dx.doi.org/10.3389/fendo.2021.642857
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