ESR measurement of endogenous nitric oxide in liver and blood of mice subjected to hepatic ischemia-reperfusion

Fusako Takayama, Toru Egashira, Yasumitsu Yamanaka

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

To examine the role of nitric oxide (NO) in liver ischemia-reperfusion injuries, electron spin resonance (ESR) measurements were employed to directly detect NO in the liver and blood of mice subjected to liver ischemia-reperfusion. We used an NO trapping and accumulating reagent, N- (dithiocarboxy)sarcosine/Fe2+ (DTCS/Fe) that specifically binds to NO to form stable NO adduct, (DTCS)/Fe-NO. The DTCS/Fe-NO adduct is detectable by ESR spectrometry at room temperature, giving rise to a characteristic triplet spectrum. The (DTCS)/Fe-NO contents in liver and blood decreased during ischemia, and still more decreased at the early reflow phase. Then, 12 h after reflow onset, the adduct content increased approximately 1.5-fold in the liver and 2.5-fold in the blood compared to that in a non-ischemia group. The fluctuation of liver NO levels may have resulted from a lack of oxygen during ischemia and the scavenging of superoxide anions during the early reflow phase. Therefore, NO levels would result in decline, then a subsequent boost in NO levels as cells respond to the reperfusion-induced damage. Blood endotoxin concentration and serum aspartate oxoglutarate aminotransferase (AST), alanine oxoglutarate aminotransferase (ALT), and lactate dehydrogenase (LDH) levels were significantly higher in the liver ischemia-reperfused group than in the sham-operated group. Pretreatment with N(G)-mono-N-methyl-L- arginine, an NO synthase inhibitor resulted in decreased NO levels to an undetectable level, all the more increased lipid peroxide contents in liver, and the amelioration of hepatic injury induced by 60 min of liver ischemia followed by 60 min of reperfusion. From these results, NO was interpreted to protect against injuries in part to brake the progression of the free radical chain reaction.

Original languageEnglish
Pages (from-to)45-51
Number of pages7
JournalPathophysiology
Volume6
Issue number1
DOIs
Publication statusPublished - Apr 1999
Externally publishedYes

Fingerprint

Electron Spin Resonance Spectroscopy
Reperfusion
Nitric Oxide
Ischemia
Liver
Ketoglutaric Acids
Reperfusion Injury
Sarcosine
Lipid Peroxides
Wounds and Injuries
Aspartate Aminotransferases
Alanine Transaminase
L-Lactate Dehydrogenase
Endotoxins
Nitric Oxide Synthase
Superoxides
Free Radicals
Arginine
Spectrum Analysis

Keywords

  • ESR
  • Free radical
  • Iron complex-NO
  • Ischemia-reperfusion
  • Nitric oxide
  • Phosphatidylcholine hydroperoxide

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

ESR measurement of endogenous nitric oxide in liver and blood of mice subjected to hepatic ischemia-reperfusion. / Takayama, Fusako; Egashira, Toru; Yamanaka, Yasumitsu.

In: Pathophysiology, Vol. 6, No. 1, 04.1999, p. 45-51.

Research output: Contribution to journalArticle

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