Measurement of nitric oxide and peroxynitrite generation in the postischemic heart

Evidence for peroxynitrite-mediated reperfusion injury

Penghai Wang, Jay L. Zweier

Research output: Contribution to journalArticle

Abstract

Altered nitric oxide (NO(·)) production is a critical factor in tissue reperfusion injury; however, controversy remains regarding these alterations and how they cause injury. Since superoxide (O 2/ -·) generation is triggered during the early period of reperfusion the cytotoxic oxidant peroxynitrite (ONOO -) could be formed, but it is not known if this occurs. Therefore electron paramagnetic resonance and chemiluminescence studies were performed of the magnitude and time course of NO(·), O 2/ -·, and ONOO - formation in the postischemic heart. Isolated rat hearts were subjected either to normal perfusion or to reperfusion after 30 min of ischemia in the presence of the NO(·) trap Fe 2+-N-methyl-D-glucamine dithiocarbamate with electron paramagnetic resonance measurements performed on the effluent. Although only trace signals were present prior to ischemia, prominent NO(·) adduct signals were seen during the first 2 min of reflow which were abolished by nitric oxide synthase (NOS) inhibition. Similar studies with the O 2/ -· trap 5,5-dimethyl-1-pyrroline N-oxide demonstrated a burst of O 2/ - · generation over the first 2 min of reflow. Chemiluminescence measurements using 5-amino-2,3-dihydro-1,4-phthalazinedione (luminol) demonstrated a similar marked increase in ONOO - which was blocked by NOS inhibitors or superoxide dismutase. NOS inhibition or superoxide dismutase greatly enhanced the recovery of contractile function in postischemic hearts. Immunohistology demonstrated that the ONOO --mediated nitration product nitrotyrosine was formed in postischemic hearts but not in normally perfused controls. Thus, NO(·) formation is increased during the early period of reflow and reacts with O 2/ -· to form ONOO -, which results in amino acid nitration and cellular injury.

Original languageEnglish (US)
Pages (from-to)29223-29230
Number of pages8
JournalJournal of Biological Chemistry
Volume271
Issue number46
DOIs
StatePublished - 1996

Fingerprint

Peroxynitrous Acid
Reperfusion Injury
Nitric Oxide
Nitric Oxide Synthase
Nitration
Luminol
Chemiluminescence
Electron Spin Resonance Spectroscopy
Superoxide Dismutase
Reperfusion
Paramagnetic resonance
Ischemia
Luminescent Measurements
Recovery of Function
Wounds and Injuries
Thromboplastin
Luminescence
Oxidants
Superoxides
Rats

ASJC Scopus subject areas

  • Biochemistry

Cite this

Measurement of nitric oxide and peroxynitrite generation in the postischemic heart : Evidence for peroxynitrite-mediated reperfusion injury. / Wang, Penghai; Zweier, Jay L.

In: Journal of Biological Chemistry, Vol. 271, No. 46, 1996, p. 29223-29230.

Research output: Contribution to journalArticle

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