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

• Published in: Experimental cell research 2016-01, Vol.340 (1), p.139-149
• Main Authors: Ha, Tae-Sun, Park, Hye-Young, Seong, Su-Bin, Ahn, Hee Yul
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; ABUNDANCE; ADENINES; ALBUMINS; Animals; ASCORBIC ACID; CATTLE; Cell Membrane Permeability - drug effects; Cells, Cultured; Cellular biology; CONCENTRATION RATIO; DIAPHRAGM; Dose-Response Relationship, Drug; DOSES; ELECTRON SCANNING; ESTERS; FLUORIDES; IN VITRO; INDUCTION; MESSENGER-RNA; MICE; NADP; NADPH oxidase; NADPH Oxidase 4; NADPH Oxidases - genetics; Nephrotic syndrome; NICOTINAMIDE; OXIDASES; OXIDATION; Oxidative stress; Oxidative Stress - drug effects; OXYGEN; PERMEABILITY; Podocyte; Podocytes - cytology; Podocytes - drug effects; Podocytes - metabolism; Proteinuria; PUROMYCIN; Puromycin aminonucleoside; Puromycin Aminonucleoside - pharmacology; RATS; RNA, Messenger - genetics; RNA, Messenger - metabolism; SCANNING ELECTRON MICROSCOPY; Structure-Activity Relationship; Studies; ULTRASTRUCTURAL CHANGES; ZO-1; Zonula Occludens-1 Protein - genetics; Zonula Occludens-1 Protein - metabolism
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2015.12.001

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. [Display omitted] •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.