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KMID : 1130320080510010054
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Korean Journal of Pediatrics
2008 Volume.51 No. 1 p.54 ~ p.61
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Effects of puromycin aminonucleoside on the cytoskeletal changes of glomerular epithelial cells
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Lee Jun-Ho
Ha Tae-Sun
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Abstract
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Purpose: This study was designed to clarify the mechanism of proteinuria in nephrotic syndrome
patients by using puromycin aminonucleoside (PAN) nephrosis model.
Methods: Following administration of various concentrations of PAN and antioxidants we observed the changes of podocyte cytoskeletons in cultured rat glomerular epithelial cells (GEpC) by method of scanning electron microscope, reactive oxyten species (ROS) analysis, permeability assay, confocal microscope, and Western blot assay.
Results: PAN not only induced the ultrastructural changes of GEpC, such as shortening and fusion of microvilli, but also separated the intercellular gaps and linear ZO-1. PAN induced oxidative stresses in time and dose dependent manners and increases of intercellular permeability in anti-oxidants inhibitable manners. High concentration of PAN induced not only actin polymerization and disorganization, but also the conglomerulation and internal dislocation of α-actinin protein. The intensities of fluorescences of ZO-1 protein were diminished and internalized by PAN in a dose-dependent manner, which were inhibited by anti anti-oxidants.
Conclusions: PAN induced the changes of podocytes cytoskeleton and junctional barriers by way of increasing ROS in GEpC that resulted in increasing their permeability in a antioxidatn-inhibitable manner. Glomerular yperpermeability induced by PAN mediateing through oxidative stresses is thought to take part in the mechanism of proteinuria in nephrotic syndrome. (Korean J Pediatr 2008;51:54-61)
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KEYWORD
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Puromycin aminoglycoside, Nephrotic syndrome, Proteinuria, Oxidative stresses, Antioxidants
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