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A Flow Cytometry-based Assay for Measuring Mitochondrial Membrane Potential in Cardiac Myocytes After Hypoxia/Reoxygenation
Published on: July 13, 2018
MitoTEMPOL, a mitochondrial targeted antioxidant, prevents sepsis-induced diaphragm dysfunction
Gerald S Supinski1, Lin Wang1, Elizabeth A Schroder1
1Division of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, University of Kentucky, Lexington, Kentucky.
Abstract:
Clinical studies indicate that sepsis-induced diaphragm dysfunction is a major contributor to respiratory failure in mechanically ventilated patients. Currently there is no drug to treat this form of diaphragm weakness. Sepsis-induced muscle dysfunction is thought to be triggered by excessive mitochondrial free radical generation; we therefore hypothesized that therapies that target mitochondrial free radical production may prevent sepsis-induced diaphragm weakness. The present study determined whether MitoTEMPOL, a mitochondrially targeted free radical scavenger, could reduce sepsis-induced diaphragm dysfunction. Using an animal model of sepsis, we compared four groups of mice: 1) sham-operated controls, 2) animals with sepsis induced by cecal ligation puncture (CLP), 3) sham controls given MitoTEMPOL (10 mg·kg-1·day-1 ip), and 4) CLP animals given MitoTEMPOL. At 48 h after surgery, we measured diaphragm force generation, mitochondrial function, proteolytic enzyme activities, and myosin heavy chain (MHC) content. We also examined the effects of delayed administration of MitoTEMPOL (by 6 h) on CLP-induced diaphragm weakness. The effects of MitoTEMPOL on cytokine-mediated alterations on muscle cell superoxide generation and cell size in vitro were also assessed. Sepsis markedly reduced diaphragm force generation. Both immediate and delayed MitoTEMPOL administration prevented sepsis-induced diaphragm weakness. MitoTEMPOL reversed sepsis-mediated reductions in mitochondrial function, activation of proteolytic pathways, and decreases in MHC content. Cytokines increased muscle cell superoxide generation and decreased cell size, effects that were ablated by MitoTEMPOL. MitoTEMPOL and other compounds that target mitochondrial free radical generation may be useful therapies for sepsis-induced diaphragm weakness.
Insights
MitoTEMPOL, a free radical scavenger, prevents sepsis-induced diaphragm weakness and respiratory failure in mice. This mitochondrial therapy improves muscle function and may offer a new treatment for sepsis patients.
Area of Science:
- Biomedical research
- Critical care medicine
- Mitochondrial biology
Background:
- Sepsis-induced diaphragm dysfunction contributes to respiratory failure in ventilated patients.
- No current drug treatments exist for this condition.
- Excessive mitochondrial free radicals are implicated in sepsis-induced muscle dysfunction.
Purpose of the Study:
- To determine if MitoTEMPOL, a mitochondrial free radical scavenger, can prevent sepsis-induced diaphragm dysfunction.
- To investigate the therapeutic potential of targeting mitochondrial free radicals for diaphragm weakness.
Main Methods:
- Utilized a mouse model of sepsis (cecal ligation puncture).
- Compared four groups: sham, sepsis, sham + MitoTEMPOL, sepsis + MitoTEMPOL.
- Assessed diaphragm force, mitochondrial function, proteolytic enzymes, and myosin heavy chain content.
- Examined immediate and delayed MitoTEMPOL administration effects, plus in vitro cytokine effects.
Main Results:
- Sepsis significantly reduced diaphragm force generation.
- Both immediate and delayed MitoTEMPOL administration prevented sepsis-induced diaphragm weakness.
- MitoTEMPOL improved mitochondrial function, proteolytic pathways, and myosin heavy chain content.
- MitoTEMPOL counteracted cytokine-induced increases in muscle cell superoxide generation and cell size.
Conclusions:
- MitoTEMPOL effectively prevents and potentially reverses sepsis-induced diaphragm weakness in an animal model.
- Targeting mitochondrial free radical generation with compounds like MitoTEMPOL shows promise for treating sepsis-induced diaphragm dysfunction.
- This research suggests a potential new therapeutic strategy for respiratory failure in sepsis.
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