Puromycin aminonucleoside increases podocyte permeability by modulating ZO-1 in an oxidative stress-dependent manner

Journal Article · Fri Jan 01 04:00:00 EST 2016 · Experimental Cell Research

DOI:https://doi.org/10.1016/J.YEXCR.2015.12.001· OSTI ID:22746400

Ha, Tae-Sun ^[1]; Park, Hye-Young; Seong, Su-Bin ^[1]; Ahn, Hee Yul ^[2]

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1. Department of Pediatrics, College of Medicine, Chungbuk National University, Cheongju (Korea, Republic of)
2. Department of Pharmacology, College of Medicine, Chungbuk National University, Cheongju (Korea, Republic of)

Puromycin aminonucleoside (PAN)-induced nephrosis is a widely studied animal model of human idiopathic nephrotic syndrome because PAN injection into rats results in increased glomerular permeability with the characteristic ultrastructural changes in podocytes similar to human nephrosis. To investigate the role of zonula occludens (ZO)-1 and oxidative stress on PAN-induced podocyte phenotypical changes and hyperpermeability in vitro, we cultured rat and mouse podocytes and treated with various concentrations of PAN. PAN treatment increased oxidative stress level of podocytes significantly with the induction of Nox4. In addition, PAN changed the ultrastructure of podocytes, such as shortening and fusion of microvilli, and the separation of intercellular gaps, which were improved by anti-oxidative vitamin C and Nox4 siRNA. PAN also disrupted the intercellular linear ZO-1 staining and induced inner cytoplasmic re-localization of ZO-1 protein, resulting in increased podocyte intercellular permeability. PAN reduced ZO-1 protein amount and mRNA expression in a dose-dependent manner, which means that PAN could also modulate ZO-1 protein transcriptionally. However, the decreased ZO-1 protein of podocytes by PAN was improved by Nox4 siRNA transfection. Furthermore, vitamin C mitigated the quantitative and distributional disturbances of ZO-1 protein caused by PAN. Our results demonstrate that the phenotypical changes of intercellular ZO-1 by oxidative stress via Nox4 likely contribute to the glomerular hyperpermeability caused by PAN. - Highlights: • PAN increases ROS and induces the ultrastructural changes in podocytes. • Podocyte intercellular hyperpermeability by PAN is via oxidative stress in vitro. • PAN induces the inner cytoplasmic relocalization of intercellular ZO-1 in podocytes. • PAN suppresses the production of ZO-1 protein at the transcriptional level. • The quantitative and distributional changes of ZO-1 by PAN are via oxidative stress.

• OSTI ID: 22746400
• Journal Information: Experimental Cell Research, Journal Name: Experimental Cell Research Journal Issue: 1 Vol. 340; ISSN 0014-4827; ISSN ECREAL
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
ABUNDANCE
ADENINES
ALBUMINS
ASCORBIC ACID
CATTLE
CONCENTRATION RATIO
DIAPHRAGM
DOSES
ELECTRON SCANNING
ESTERS
FLUORIDES
IN VITRO
INDUCTION
MESSENGER-RNA
MICE
NADP
NICOTINAMIDE
OXIDASES
OXIDATION
OXYGEN
PERMEABILITY
PUROMYCIN
RATS
SCANNING ELECTRON MICROSCOPY
ULTRASTRUCTURAL CHANGES