EPR analysis reveals three tissues responding to endotoxin by increased formation of reactive oxygen and nitrogen species

Andrey V. Kozlov, L. Szalay, Fraz Umar, Bruno Fink, Karl Kropik, Hans Nohl, Heinz Redl, Soheyl Bahrami

Research output: Contribution to journalArticle

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Abstract

The excessive formation of reactive oxygen and nitrogen species (RONS) in tissue has been implicated in the development of various diseases. In this study we adopted ex vivo low temperature EPR spectroscopy combined with spin trapping technique to measure local RONS levels in frozen tissue samples. CP-H (1-hydroxy-3-carboxy-pyrrolidine), a new nontoxic spin probe, was used to analyze RONS in vivo. In addition, nitrosyl complexes of hemoglobin were determined to trace nitric oxide released into blood. By this technique we found that RONS formation in tissue of control animals increased in the following order: liver <heart <brain <cerebellum <lung <muscle <blood <ileum <kidney <duodenum <jejunum. We also found that endotoxin challenge, which represents the most common model of septic shock, increased the formation of RONS in rat liver, heart, lung, and blood, but decreased RONS formation in jejunum. We did not find changes in RONS levels in other parts of gut, brain, skeletal muscles, and kidney. Scavenging of RONS by CP-H was accompanied by an increase in blood pressure, indicating that LPS-induced vasodilatation may be due to RONS, but not due to nitric oxide. Experiments with tissue homogenates incubated in vitro with CP-H showed that ONOO- and O2 ·-, as well as other not identified RONS, are detectable by CP-H in tissue. In summary, low-temperature EPR combined with CP-H infusion allowed detection of local RONS formation in tissues. Increased formation of RONS in response to endotoxin challenge is organ specific.

Original languageEnglish
Pages (from-to)1555-1562
Number of pages8
JournalFree Radical Biology and Medicine
Volume34
Issue number12
DOIs
Publication statusPublished - Jun 15 2003

Fingerprint

Reactive Nitrogen Species
Endotoxins
Paramagnetic resonance
Reactive Oxygen Species
Tissue
Blood
Jejunum
Liver
Muscle
Brain
Nitric Oxide
Spin Trapping
Kidney
Lung
Temperature
Scavenging
Blood pressure
Septic Shock
Ileum
Duodenum

Keywords

  • 1-Hydroxy-3-carboxy-pyrrolidine
  • Blood
  • Brain
  • Electron paramagnetic resonance
  • Endotoxin
  • Free radicals
  • Heart
  • Intestine
  • Liver
  • Lung
  • Muscle
  • Oxidative stress

ASJC Scopus subject areas

  • Medicine(all)
  • Toxicology
  • Clinical Biochemistry

Cite this

EPR analysis reveals three tissues responding to endotoxin by increased formation of reactive oxygen and nitrogen species. / Kozlov, Andrey V.; Szalay, L.; Umar, Fraz; Fink, Bruno; Kropik, Karl; Nohl, Hans; Redl, Heinz; Bahrami, Soheyl.

In: Free Radical Biology and Medicine, Vol. 34, No. 12, 15.06.2003, p. 1555-1562.

Research output: Contribution to journalArticle

Kozlov, Andrey V. ; Szalay, L. ; Umar, Fraz ; Fink, Bruno ; Kropik, Karl ; Nohl, Hans ; Redl, Heinz ; Bahrami, Soheyl. / EPR analysis reveals three tissues responding to endotoxin by increased formation of reactive oxygen and nitrogen species. In: Free Radical Biology and Medicine. 2003 ; Vol. 34, No. 12. pp. 1555-1562.
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