Inhibition of nitric oxide synthase ameliorates cellular injury in sickle cell mouse kidneys

Norman Bank, Militza Kiroycheva, Pravin C. Singhal, Gillian M. Anthony, Garry J. Southan, Csaba Szabo

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Background: In previous studies of transgenic sickle cell mice, increased renal expression of inducible nitric oxide synthase (iNOS) and endothelial cell isoform of NOS (EcNOS) was found by Western blot and immunohistochemistry. In addition, putative evidence of peroxynitrite (ONOO- ) formation was found in the form of positive immunostaining and immunoblot for nitrotyrosine. Apoptosis was also detected by DNA strand breakage and TUNEL assay. The present study was carried out to examine the role of NO/ONOO- in mediating renal tubular cell apoptosis in sickle cell mouse kidneys. Methods: Mercaptoethylguanidine (MEG), a compound that selectively inhibits iNOS and also is a scavenger of ONOO-, was administered intraperitoneally over a five-day period to control and β(s) mice. Immunohistochemistry of iNOS and nitrotyrosine, DNA electrophoresis, ApoTACS assay for apoptosis, and Western blot of poly(ADP-ribose) polymerase (PARP) were carried out. Results: MEG administration virtually eliminated renal immunostaining of iNOS and nitrotyrosine and prevented DNA strand breakage. In addition, Western blot analysis of PARP, a nuclear DNA-reparative enzyme activated in response to DNA strand breakage, was found to be cleavaged in hypoxic β(s) mice, but was partially protected in MEG-treated β(s) hypoxic mice. Finally, apoptosis was markedly reduced by MEG in β(s) hypoxic mice. Conclusions: These observations provide evidence that NO and/or ONOO- are responsible for initiating cell damage, which leads to apoptosis in sickle cell mouse kidneys.

Original languageEnglish (US)
Pages (from-to)82-89
Number of pages8
JournalKidney International
Volume58
Issue number1
DOIs
StatePublished - 2000
Externally publishedYes

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Nitric Oxide Synthase
Kidney
Nitric Oxide Synthase Type II
Wounds and Injuries
Apoptosis
DNA
Poly(ADP-ribose) Polymerases
Western Blotting
Immunohistochemistry
Peroxynitrous Acid
In Situ Nick-End Labeling
Electrophoresis
Protein Isoforms
Endothelial Cells
2-mercaptoethylguanidine
Enzymes
3-nitrotyrosine

Keywords

  • Apoptosis
  • ApoTACS assay
  • DNA strand breakage
  • Hypoxia
  • Immunohistochemistry
  • Poly(ADP-ribose) polymerase

ASJC Scopus subject areas

  • Nephrology

Cite this

Inhibition of nitric oxide synthase ameliorates cellular injury in sickle cell mouse kidneys. / Bank, Norman; Kiroycheva, Militza; Singhal, Pravin C.; Anthony, Gillian M.; Southan, Garry J.; Szabo, Csaba.

In: Kidney International, Vol. 58, No. 1, 2000, p. 82-89.

Research output: Contribution to journalArticle

Bank, Norman ; Kiroycheva, Militza ; Singhal, Pravin C. ; Anthony, Gillian M. ; Southan, Garry J. ; Szabo, Csaba. / Inhibition of nitric oxide synthase ameliorates cellular injury in sickle cell mouse kidneys. In: Kidney International. 2000 ; Vol. 58, No. 1. pp. 82-89.
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AU - Szabo, Csaba

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