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Heat shock protein 70/nitric oxide effect on stretched tubular epithelial cells linked to WT-1 cytoprotection during neonatal obstructive nephropathy

  • Nephrology - Original Paper
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Abstract

Background

Mechanical stress is a key pathogenic driver of apoptosis in the tubular epithelium in obstructive nephropathy. Heat shock protein 70 (Hsp70) and Wilms’ tumor (WT-1) have been proposed to represent linked downstream effectors of the cytoprotective properties of NO. In the present study, we sought to evaluate whether the cytoprotective effects of l-arginine in neonatal obstructive nephropathy may be associated with NO-dependent increases in WT-1 and Hsp70 expression.

Methods

Neonatal Wistar–Kyoto rats were submitted to complete unilateral ureteral obstruction (UUO) and treated thereafter with vehicle, L-NAME or l-arginine by daily gavage for 14 days to block or augment NO levels, respectively. Normal rat kidney epithelial cells by NRK-52E were exposed to mechanical stress in vitro in the presence or absence of L-NAME, l-arginine, sodium nitroprusside (SNP), l-arginine + SNP or l-arginine/L-NAME. Induction of apoptosis and the mRNA expression of WT-1 and Hsp70 genes were assessed.

Results

WT-1 and Hsp70 genes expression decreased in the presence of L-NAME and following UUO coincident with increased tubular apoptosis. l-arginine treatment increased NO levels, reduced apoptosis and restored expression levels of WT-1 and Hsp70 to control levels. l-arginine treatment in vitro reduced basal apoptotic rates and prevented apoptosis in response to mechanical strain, an effect enhanced by SNP co-incubation. L-NAME increased apoptosis and prevented the anti-apoptotic action of l-arginine.

Conclusions

l-arginine treatment in experimental neonatal UUO reduces apoptosis coincident with restoration of WT-1 and Hsp70 expression levels and directly inhibits mechanical strain-induced apoptosis in an NO-dependent manner in vitro. This potentially implicates an NO-Hsp70-WT-1 axis in the cytoprotective effects of l-arginine.

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Funding

This work was supported by Grants from the Research and Technology Council of Cuyo University (SECyT), Mendoza, Argentina, and PICT 2012-0234.

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Authors and Affiliations

  1. Laboratorio de Farmacología Experimental Básica y Traslacional. IMBECU-CONICET (National Council of Scientific and Technical Research of Argentina), Buenos Aires, Argentina

    Luciana Mazzei, Fernando Darío Cuello-Carrión & Walter Manucha

  2. Área de Farmacología, Departamento de Patología, Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, Mendoza, Argentina

    Luciana Mazzei & Walter Manucha

  3. Conway Institute of Biomolecular and Biomedical Research, School of Medicine, University College Dublin, Dublin, Ireland

    Neil Docherty

  4. Pharmacology Area, Pathology Department, Medical Sciences College, National University of Cuyo, Mendoza, CP5500, Argentina

    Walter Manucha

Authors
  1. Luciana Mazzei
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  2. Fernando Darío Cuello-Carrión
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  3. Neil Docherty
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  4. Walter Manucha
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Correspondence to Walter Manucha.

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Ethical statement

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted (IRB approval number 48/2015). This article does not contain any studies with human participants performed by any of the authors.

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Mazzei, L., Cuello-Carrión, F.D., Docherty, N. et al. Heat shock protein 70/nitric oxide effect on stretched tubular epithelial cells linked to WT-1 cytoprotection during neonatal obstructive nephropathy. Int Urol Nephrol 49, 1875–1892 (2017). https://doi.org/10.1007/s11255-017-1658-z

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  • DOI: https://doi.org/10.1007/s11255-017-1658-z

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