title: Puromycin aminonucleoside increases podocyte permeability by modulating ZO-1 in an oxidative stress-dependent manner.
abstract: 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.
authors.affiliation: Department of Pediatrics, College of Medicine, Chungbuk National University, Cheongju, South Korea. Electronic address: tsha@chungbuk.ac.kr.
authors.forename: Tae-Sun
authors.initials: TS
authors.lastname: Ha
authors.affiliation: Department of Pediatrics, College of Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Hye-Young
authors.initials: HY
authors.lastname: Park
authors.affiliation: Department of Pediatrics, College of Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Su-Bin
authors.initials: SB
authors.lastname: Seong
authors.affiliation: Department of Pharmacology, College of Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Hee Yul
authors.initials: HY
authors.lastname: Ahn
mesh_terms: D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D051379:Mice; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000235:genetics; D018384:Oxidative Stress / Q000187:drug effects*; D050199:Podocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011692:Puromycin Aminonucleoside / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D013329:Structure-Activity Relationship; D062826:Zonula Occludens-1 Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D012333:RNA, Messenger; C569043:Tjp1 protein, mouse; C569044:Tjp1 protein, rat; D062826:Zonula Occludens-1 Protein; D011692:Puromycin Aminonucleoside; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; C422943:Nox4 protein, mouse
keywords: NADPH oxidase; Nephrotic syndrome; Oxidative stress; Podocyte; Proteinuria; Puromycin aminonucleoside; ZO-1