Puromycin aminonucleoside modulates p130Cas of podocytes.
10.3345/kjp.2012.55.10.371
- Author:
Tae Sun HA
1
;
Ji Young CHOI
;
Hye Young PARK
Author Information
1. Department of Pediatrics, Chungbuk National University College of Medicine, Cheongju, Korea. tsha@chungbuk.ac.kr
- Publication Type:Original Article ; In Vitro
- Keywords:
Cytoskeleton;
p130Cas;
Podocytes cell;
Puromycin aminonucleoside
- MeSH:
Actin Cytoskeleton;
Actins;
Animals;
Antioxidants;
Ascorbic Acid;
Blotting, Western;
Cadherins;
Catechin;
Crk-Associated Substrate Protein;
Cytoplasm;
Cytoskeleton;
Diaphragm;
Epithelial Cells;
Fluorescent Antibody Technique;
Foot;
Glomerular Basement Membrane;
Mice;
Oxidative Stress;
Podocytes;
Probucol;
Proteins;
Puromycin;
Puromycin Aminonucleoside;
Rats;
RNA, Messenger
- From:Korean Journal of Pediatrics
2012;55(10):371-376
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: Puromycin aminonucleoside (PAN) specifically injures podocytes, leading to foot process effacement, actin cytoskeleton disorganization, and abnormal distribution of slit diaphragm proteins. p130Cas is a docking protein connecting F-actin fibers to the glomerular basement membrane (GBM) and adapter proteins in glomerular epithelial cells (GEpCs; podocytes). We investigated the changes in the p130Cas expression level in the PAN-induced pathological changes of podocytes in vitro. METHODS: We observed changes in the p130Cas expression in cultured rat GEpCs and mouse podocytes treated with various concentrations of PAN and antioxidants, including probucol, epigallocatechin gallate (EGCG), and vitamin C. The changes in the p130Cas expression level were analyzed using confocal immunofluorescence imaging, Western blotting, and polymerase chain reaction. RESULTS: In the immunofluorescence study, p130Cas showed a diffuse cytoplasmic distribution with accumulation at distinct sites visible as short stripes and colocalized with P-cadherin. The fluorescences of the p130Cas protein were internalized and became granular by PAN administration in a dose-dependent manner, which had been restored by antioxidants, EGCG and vitamin C. PAN also decreased the protein and mRNA expression levels of p130Cas at high doses and in a longer exposed duration, which had been also reversed by antioxidants. CONCLUSION: These findings suggest that PAN modulates the quantitative and distributional changes of podocyte p130Cas through oxidative stress resulting in podocyte dysfunction.
