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Denaturing Urea Polyacrylamide Gel Electrophoresis Urea PAGE
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Urea Memory: Transient Cell Exposure to Urea Causes Persistent Mitochondrial ROS Production and Endothelial
Maria d'Apolito1, Anna Laura Colia2, Enrica Manca3
1Pediatric Research Center, Department of Medical and Surgical Sciences, University of Foggia, 71122 Foggia, Italy. maria.dapolito@unifg.it.
Toxins
|October 14, 2018
Summary
Elevated urea causes sustained reactive oxygen species (ROS) production in cells, even after urea removal. This persistent ROS contributes to kidney damage by affecting mitochondrial function and inflammation.
Area of Science:
- Biochemistry
- Cell Biology
- Nephrology
Background:
- Elevated urea levels post-dialysis are known to induce reactive oxygen species (ROS) in various cell types.
- The persistence and origin of ROS production after urea removal remain unclear.
Purpose of the Study:
- To investigate if urea-induced ROS production persists after urea removal.
- To characterize the origin and effects of sustained ROS in human arterial endothelial cells.
Main Methods:
- Human arterial endothelial cells were incubated with urea, followed by incubation in urea-free medium.
- Assessed ROS levels, mitochondrial DNA (mtDNA) copy number, and expression of mitochondrial and inflammatory markers.
- Utilized manganese superoxide dismutase (MnSOD) expression, Nox1/4 inhibition (GKT13781), and SOD-catalase mimetic (MnTBAP) for intervention.
Main Results:
- Sustained elevated ROS levels were observed even after urea removal.
- Persistent ROS reduced mtDNA copy number, electron transport chain transcripts, and mitochondrial fusion proteins, while increasing mitochondrial fission proteins.
- Inflammatory markers were persistently expressed, and these effects were reversed by MnTBAP.
Conclusions:
- Urea-induced ROS production can persist after urea removal, contributing to endothelial dysfunction.
- Sustained ROS impacts mitochondrial integrity and promotes inflammation, potentially driving kidney damage despite dialysis.
- Targeting persistent ROS may offer therapeutic strategies for kidney disease progression.
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