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KMID : 0811820130170020079
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Journal of Korean Society of Pediatric Nephrology
2013 Volume.17 No. 2 p.79 ~ p.85
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Effect of Puromycin Aminonucleoside on Podocyte P-Cadherin
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Ha Tae-Sun
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Abstract
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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 μg/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 μg/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.
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KEYWORD
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P-Cadherin, Puromycin aminonucleoside (PAN)-induced nephropathy, Glomerular epithelial cells (GEpC), Podocyte
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