Sepsis Disrupts Mitochondrial Function and Diaphragm Morphology
BackgroundThe diaphragm is the primary muscle of inspiration, and its dysfunction is frequent during sepsis. However, the mechanisms associated with sepsis and diaphragm dysfunction are not well understood. In this study, we evaluated the morphophysiological changes of the mitochondrial diaphragm 5...
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Frontiers Media S.A.
2021-09-01
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Online Access: | https://www.frontiersin.org/articles/10.3389/fphys.2021.704044/full |
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doaj-41d4ed5bc4ec468d9797eaa172f3ef4a |
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Article |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Thamires Siqueira Oliveira Anderson Teixeira Santos Cherley Borba Vieira Andrade Johnatas Dutra Silva Natália Blanco Nazareth de Novaes Rocha Juliana Woyames Pedro Leme Silva Patricia Rieken Macedo Rocco Wagner Seixas da-Silva Tânia Maria Ortiga-Carvalho Flavia Fonseca Bloise |
spellingShingle |
Thamires Siqueira Oliveira Anderson Teixeira Santos Cherley Borba Vieira Andrade Johnatas Dutra Silva Natália Blanco Nazareth de Novaes Rocha Juliana Woyames Pedro Leme Silva Patricia Rieken Macedo Rocco Wagner Seixas da-Silva Tânia Maria Ortiga-Carvalho Flavia Fonseca Bloise Sepsis Disrupts Mitochondrial Function and Diaphragm Morphology Frontiers in Physiology diaphragm sepsis mitochondria CLP oxidative phosphorylation muscle |
author_facet |
Thamires Siqueira Oliveira Anderson Teixeira Santos Cherley Borba Vieira Andrade Johnatas Dutra Silva Natália Blanco Nazareth de Novaes Rocha Juliana Woyames Pedro Leme Silva Patricia Rieken Macedo Rocco Wagner Seixas da-Silva Tânia Maria Ortiga-Carvalho Flavia Fonseca Bloise |
author_sort |
Thamires Siqueira Oliveira |
title |
Sepsis Disrupts Mitochondrial Function and Diaphragm Morphology |
title_short |
Sepsis Disrupts Mitochondrial Function and Diaphragm Morphology |
title_full |
Sepsis Disrupts Mitochondrial Function and Diaphragm Morphology |
title_fullStr |
Sepsis Disrupts Mitochondrial Function and Diaphragm Morphology |
title_full_unstemmed |
Sepsis Disrupts Mitochondrial Function and Diaphragm Morphology |
title_sort |
sepsis disrupts mitochondrial function and diaphragm morphology |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Physiology |
issn |
1664-042X |
publishDate |
2021-09-01 |
description |
BackgroundThe diaphragm is the primary muscle of inspiration, and its dysfunction is frequent during sepsis. However, the mechanisms associated with sepsis and diaphragm dysfunction are not well understood. In this study, we evaluated the morphophysiological changes of the mitochondrial diaphragm 5 days after sepsis induction.MethodsMale C57Bl/6 mice were divided into two groups, namely, cecal ligation and puncture (CLP, n = 26) and sham-operated (n = 19). Mice received antibiotic treatment 8 h after surgery and then every 24 h until 5 days after surgery when mice were euthanized and the diaphragms were collected. Also, diaphragm function was evaluated in vivo by ultrasound 120 h after CLP. The tissue fiber profile was evaluated by the expression of myosin heavy chain and SERCA gene by qPCR and myosin protein by using Western blot. The Myod1 and Myog expressions were evaluated by using qPCR. Diaphragm ultrastructure was assessed by electron microscopy, and mitochondrial physiology was investigated by high-resolution respirometry, Western blot, and qPCR.ResultsCecal ligation and puncture mice developed moderated sepsis, with a 74% survivor rate at 120 h. The diaphragm mass did not change in CLP mice compared with control, but we observed sarcomeric disorganization and increased muscle thickness (38%) during inspiration and expiration (21%). Septic diaphragm showed a reduction in fiber myosin type I and IIb mRNA expression by 50% but an increase in MyHC I and IIb protein levels compared with the sham mice. Total and healthy mitochondria were reduced by 30% in septic mice, which may be associated with a 50% decrease in Ppargc1a (encoding PGC1a) and Opa1 (mitochondria fusion marker) expressions in the septic diaphragm. The small and non-functional OPA1 isoform also increased 70% in the septic diaphragm. These data suggest an imbalance in mitochondrial function. In fact, we observed downregulation of all respiratory chain complexes mRNA expression, decreased complex III and IV protein levels, and reduced oxygen consumption associated with ADP phosphorylation (36%) in CLP mice. Additionally, the septic diaphragm increased proton leak and downregulated Sod2 by 70%.ConclusionThe current model of sepsis induced diaphragm morphological changes, increased mitochondrial damage, and induced functional impairment. Thus, diaphragm damage during sepsis seems to be associated with mitochondrial dysfunction. |
topic |
diaphragm sepsis mitochondria CLP oxidative phosphorylation muscle |
url |
https://www.frontiersin.org/articles/10.3389/fphys.2021.704044/full |
work_keys_str_mv |
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doaj-41d4ed5bc4ec468d9797eaa172f3ef4a2021-09-07T04:30:04ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2021-09-011210.3389/fphys.2021.704044704044Sepsis Disrupts Mitochondrial Function and Diaphragm MorphologyThamires Siqueira Oliveira0Anderson Teixeira Santos1Cherley Borba Vieira Andrade2Johnatas Dutra Silva3Natália Blanco4Nazareth de Novaes Rocha5Juliana Woyames6Pedro Leme Silva7Patricia Rieken Macedo Rocco8Wagner Seixas da-Silva9Tânia Maria Ortiga-Carvalho10Flavia Fonseca Bloise11Laboratory of Translational Endocrinology, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Metabolic Adaptations, Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Translational Endocrinology, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Pulmonary Investigation, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Pulmonary Investigation, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilPhysiology and Pharmacology Department, Biomedical Institute, Fluminense Federal University, Niteroi, BrazilLaboratory of Molecular Endocrinology, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Pulmonary Investigation, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Pulmonary Investigation, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Metabolic Adaptations, Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Translational Endocrinology, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Translational Endocrinology, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, BrazilBackgroundThe diaphragm is the primary muscle of inspiration, and its dysfunction is frequent during sepsis. However, the mechanisms associated with sepsis and diaphragm dysfunction are not well understood. In this study, we evaluated the morphophysiological changes of the mitochondrial diaphragm 5 days after sepsis induction.MethodsMale C57Bl/6 mice were divided into two groups, namely, cecal ligation and puncture (CLP, n = 26) and sham-operated (n = 19). Mice received antibiotic treatment 8 h after surgery and then every 24 h until 5 days after surgery when mice were euthanized and the diaphragms were collected. Also, diaphragm function was evaluated in vivo by ultrasound 120 h after CLP. The tissue fiber profile was evaluated by the expression of myosin heavy chain and SERCA gene by qPCR and myosin protein by using Western blot. The Myod1 and Myog expressions were evaluated by using qPCR. Diaphragm ultrastructure was assessed by electron microscopy, and mitochondrial physiology was investigated by high-resolution respirometry, Western blot, and qPCR.ResultsCecal ligation and puncture mice developed moderated sepsis, with a 74% survivor rate at 120 h. The diaphragm mass did not change in CLP mice compared with control, but we observed sarcomeric disorganization and increased muscle thickness (38%) during inspiration and expiration (21%). Septic diaphragm showed a reduction in fiber myosin type I and IIb mRNA expression by 50% but an increase in MyHC I and IIb protein levels compared with the sham mice. Total and healthy mitochondria were reduced by 30% in septic mice, which may be associated with a 50% decrease in Ppargc1a (encoding PGC1a) and Opa1 (mitochondria fusion marker) expressions in the septic diaphragm. The small and non-functional OPA1 isoform also increased 70% in the septic diaphragm. These data suggest an imbalance in mitochondrial function. In fact, we observed downregulation of all respiratory chain complexes mRNA expression, decreased complex III and IV protein levels, and reduced oxygen consumption associated with ADP phosphorylation (36%) in CLP mice. Additionally, the septic diaphragm increased proton leak and downregulated Sod2 by 70%.ConclusionThe current model of sepsis induced diaphragm morphological changes, increased mitochondrial damage, and induced functional impairment. Thus, diaphragm damage during sepsis seems to be associated with mitochondrial dysfunction.https://www.frontiersin.org/articles/10.3389/fphys.2021.704044/fulldiaphragmsepsismitochondriaCLPoxidative phosphorylationmuscle |