Effect of Puromycin Aminonucleoside on Podocyte P-Cadherin.
- Author:
Tae Sun HA
1
Author Information
1. Department of Pediatrics, College of Medicine, Chungbuk National University, Cheongju, Korea. tsha@chungbuk.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
P-Cadherin;
Puromycin aminonucleoside (PAN)-induced nephropathy;
Glomerular epithelial cells (GEpC);
Podocyte
- MeSH:
Animals;
Ascorbic Acid;
Blotting, Western;
Cadherins*;
Cytoplasm;
Diaphragm;
Epithelial Cells;
Fluorescent Antibody Technique;
Foot;
Glycyrrhetinic Acid;
Podocytes*;
Proteinuria;
Puromycin Aminonucleoside*;
Puromycin*;
Rats;
RNA, Messenger
- From:Journal of the Korean Society of Pediatric Nephrology
2013;17(2):79-85
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: To test whether the expression of P-cadherin, a component of slit diaphragms between podocyte foot processes, would be altered by puromycin aminonucleoside (PAN) in a cultured podocyte in vitro. METHODS: Rat glomerular epithelial cells (GEpC) were cultured with various concentrations of PAN. The distribution of P-cadherin was examined with a confocal microscope. Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to measure the change in P-cadherin expression. RESULTS: This study found that P-cadherin was concentrated in the inner and peripheral cytoplasm with high concentrations of PAN under immunofluorescence views. Western blotting of GEpC revealed that PAN induced a decrease of P-cadherin in dose- and time-dependent manners. A high dose (50 microg/mL) of PAN decreased P-cadherin expression by 21.9% at 24 h (P<0.05) and 31.9% at 48 h (P<0.01) compared to those without PAN. In RT-PCR, high concentrations (50 microg/mL) of PAN also decreased P-cadherin mRNA expression, similar to protein suppression, by 23.5% at 48 h (P<0.05). CONCLUSION: Podocytes exposed to PAN in vitro concentrated P-cadherin internally, and reduced P-cadherin mRNA and protein expression. This could explain the development of proteinuria in experimental PAN-induced nephropathy.
