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Journal of the Korean Society of Pediatric Nephrology 2011;15(2): 138-145. doi: https://doi.org/10.3339/jkspn.2011.15.2.138
Puromycin aminonucleoside 투여에 따른 사구체 족세포 ${beta}$-catenin의 변화
최지영, 안은미, 박혜영, 신재일, 하태선
1충북대학교 의과대학 소아과학교실
2충북대학교 의과대학 소아과학교실
3충북대학교 의과대학 소아과학교실
4연세대학교 의과대학 소아과학교실
5충북대학교 의과대학 소아과학교실
The Change of Podocyte ${beta}$-Catenin by Puromycin Aminonucleoside
Ji-Young Choi, Eun-Mi Ahn, Hye-Young Park, Jae-Il Shin, Tae-Sun Ha
1Department of Pediatrics, College of Medicine, Chungbuk National University Cheongju, The Institute of Kidney Disease
2Department of Pediatrics, College of Medicine, Chungbuk National University Cheongju, The Institute of Kidney Disease
3Department of Pediatrics, College of Medicine, Chungbuk National University Cheongju, The Institute of Kidney Disease
4Department of Pediatrics, Yonsei University College of Medicine, Severance Children's Hospital
5Department of Pediatrics, College of Medicine, Chungbuk National University Cheongju, The Institute of Kidney Disease
Received: September 1, 2011;  Accepted: October 4, 2011.
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ABSTRACT
Purpose : To test whether the expression of ${beta}$-catenin, a component of podocyte as a filtration molecule, would be altered by puromycin aminonucleoside (PAN) in the cultured podocyte in vitro.
Methods : We cultured rat glomerular epithelial cells (GEpC) with various concentrations of PAN and examined the distribution of ${beta}$-catenin by confocal microscope and measured the change of ${beta}$-catenin expression by Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR).
Results :We found that ${beta}$-catenin relocalized from peripheral cytoplasm to inner cytoplasm, therefore, intercellular separations were seen in confluently cultured cells by high concentrations of PAN in immunofluorescence views. In Western blotting of GEpC, PAN ($50{mu}g/mL$) decreased ${beta}$-catenin expression by 34.9% at 24 hrs and 34.3% at 48 hrs, compared to those in without PAN condition (P<0.05). In RT-PCR, high concentrations ($50{mu}g/mL$) of PAN also decreased ${beta}$-catenin mRNA expression similar to protein suppression by 25.4% at 24 hrs and 51.8% at 48 hrs (P<0.05).
Conclusion : Exposure of podocytes to PAN in vitro relocates ${beta}$-catenin internally and reduces ${beta}$-catenin mRNA and protein expression, which could explain the development of proteinuria in experimental PAN-induced nephropathy.
Key words: ${\beta}$-Catenin | Puromycin aminonucleoside (PAN)-induced nephropathy | Glomerular epithelial cells (GEpC) | Podocyte
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