Measurement of exhaled hydrogen peroxide from rabbit lungs

Abstract Exhaled H2O2 is considered an indicator of lung inflammatory and oxidative stress. Moreover, H2O2 may be involved in signal transduction processes. It is not fully elucidated to what extent (i) H2O2 escapes from the intravascular compartment, and (ii) pulmonary H2O2 generation and nasopharyngeal H2O2 generation contribute to exhaled H2O2. We investigated H2O2 concentrations in breath condensate from isolated bufferperfused and ventilated rabbit lungs, and from both intubated and spontaneously breathing rabbits with a horseradish peroxidase/ 2,7dichlorofluorescin assay. For the perfused lungs, a H2O2 concentration of 58±19 nM was found. Addition of H2O2 to the buffer fluid resulted in only minute appearance in the exhaled air (<0.001%). Levels of exhaled H2O2 in intubated rabbits and perfused lungs were virtually identical. Nearly tenfold higher levels were detected in spontaneously breathing rabbits. Decreasing the inspired oxygen concentration from 21% to 1% resulted in a tendency toward decreased H2O2 exhalation in perfused lungs. In contrast, phorbol-12-myristate-13- acetate (PMA) prompted a ~4-fold increase in H2O2 exhalation. We conclude that the horseradish peroxidase/ 2',7'dichlorofluorescin assay is a feasible technique to measure H2O2 in exhaled breath condensate in rabbits. When collecting exhaled air via the tracheal tube, the signal represents pulmonary H2O2 generation with the contribution of the remaining body being negligible.

[1]  D. Kettler,et al.  Hydrogen peroxide in expired breath condensate of patients with acute respiratory failure and with ARDS , 2005, Intensive Care Medicine.

[2]  N. Wilson,et al.  Dilution of respiratory solutes in exhaled condensates. , 2003, American Journal of Respiratory and Critical Care Medicine.

[3]  W. Seeger,et al.  Hypoxic pulmonary vasoconstriction: a multifactorial response? , 2001, American journal of physiology. Lung cellular and molecular physiology.

[4]  N. Chandel,et al.  Model for Hypoxic Pulmonary Vasoconstriction Involving Mitochondrial Oxygen Sensing , 2001, Circulation research.

[5]  D. Nowak,et al.  Exhalation of H2O2 and thiobarbituric acid reactive substances (TBARs) by healthy subjects. , 2001, Free radical biology & medicine.

[6]  H. Magnussen,et al.  The concentration of hydrogen peroxide in exhaled air depends on expiratory flow rate. , 2000, The European respiratory journal.

[7]  J. Lambeth,et al.  Novel homologs of gp91phox. , 2000, Trends in biochemical sciences.

[8]  W. Seeger,et al.  Hypoxic vasoconstriction in intact lungs: a role for NADPH oxidase-derived H(2)O(2)? , 2000, American journal of physiology. Lung cellular and molecular physiology.

[9]  W. Hop,et al.  Hydrogen peroxide and nitric oxide in exhaled air of children with cystic fibrosis during antibiotic treatment. , 2000, The European respiratory journal.

[10]  R. Mason,et al.  Evidence for free radical formation during the oxidation of 2'-7'-dichlorofluorescin to the fluorescent dye 2'-7'-dichlorofluorescein by horseradish peroxidase: possible implications for oxidative stress measurements. , 1999, Free radical biology & medicine.

[11]  S. Archer,et al.  O2 sensing is preserved in mice lacking the gp91 phox subunit of NADPH oxidase. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[12]  T. Pietras,et al.  Increased content of thiobarbituric acid-reactive substances and hydrogen peroxide in the expired breath condensate of patients with stable chronic obstructive pulmonary disease: no significant effect of cigarette smoking. , 1999, Respiratory medicine.

[13]  M. Vízek,et al.  Hydrogen peroxide in the breath of rats: the effects of hypoxia and paraquat. , 1999, Physiological research.

[14]  I. Horváth,et al.  Combined use of exhaled hydrogen peroxide and nitric oxide in monitoring asthma. , 1998, American journal of respiratory and critical care medicine.

[15]  N. Chandel,et al.  Mitochondrial reactive oxygen species trigger hypoxia-induced transcription. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[16]  I. Horváth,et al.  Elevated levels of expired breath hydrogen peroxide in bronchiectasis. , 1998, American journal of respiratory and critical care medicine.

[17]  W. Seeger,et al.  Nitro blue tetrazolium inhibits but does not mimic hypoxic vasoconstriction in isolated rabbit lungs. , 1998, The American journal of physiology.

[18]  Z. Kurmanowska,et al.  Increased hydrogen peroxide and thiobarbituric acid-reactive products in expired breath condensate of asthmatic patients. , 1997, The European respiratory journal.

[19]  Q. Jöbsis,et al.  Hydrogen peroxide in exhaled air is increased in stable asthmatic children. , 1997, The European respiratory journal.

[20]  B. Marshall,et al.  Pulmonary artery NADPH-oxidase is activated in hypoxic pulmonary vasoconstriction. , 1996, American journal of respiratory cell and molecular biology.

[21]  K. Aben,et al.  Increased exhalation of hydrogen peroxide in patients with stable and unstable chronic obstructive pulmonary disease. , 1996, American journal of respiratory and critical care medicine.

[22]  T. Pietras,et al.  Increased content of hydrogen peroxide in the expired breath of cigarette smokers. , 1996, The European respiratory journal.

[23]  P. Kaminski,et al.  Potential role of NADH oxidoreductase-derived reactive O2 species in calf pulmonary arterial PO2-elicited responses. , 1995, The American journal of physiology.

[24]  W. Seeger,et al.  Effects of NADPH oxidase inhibitors on hypoxic vasoconstriction in buffer-perfused rabbit lungs. , 1995, The American journal of physiology.

[25]  W. Seeger,et al.  Hypoxic vasoconstriction in buffer-perfused rabbit lungs. , 1995, Respiration physiology.

[26]  M. O’Donnell,et al.  Cerebral microvascular endothelial cell Na-K-Cl cotransport: regulation by astrocyte-conditioned medium. , 1995, The American journal of physiology.

[27]  M. Wolin,et al.  Properties of a superoxide anion-generating microsomal NADH oxidoreductase, a potential pulmonary artery PO2 sensor. , 1994, The American journal of physiology.

[28]  W. Seeger,et al.  Adult respiratory distress syndrome: model systems using isolated perfused rabbit lungs. , 1994, Methods in enzymology.

[29]  J. Crapo,et al.  Allometric relationships of cell numbers and size in the mammalian lung. , 1992, American journal of respiratory cell and molecular biology.

[30]  R. Carson,et al.  Inhibition of hypoxic pulmonary vasoconstriction by diphenyleneiodonium. , 1991, Biochemical pharmacology.

[31]  S. Archer,et al.  Simultaneous measurement of O2 radicals and pulmonary vascular reactivity in rat lung. , 1989, Journal of applied physiology.

[32]  A. Nahum,et al.  Increased hydrogen peroxide in the expired breath of patients with acute hypoxemic respiratory failure. , 1989, Chest.

[33]  Susan Budavari,et al.  The merck index an encyclopedia of chemical, drugs, and biologicals , 1989 .

[34]  M. Wolin,et al.  H2O2 and cGMP may function as an O2 sensor in the pulmonary artery. , 1989, Journal of applied physiology.

[35]  N. Voelkel.  Mechanisms of hypoxic pulmonary vasoconstriction. , 2015, The American review of respiratory disease.

[36]  W. Seeger,et al.  The glutathione redox cycle as a defense system against hydrogen-peroxide-induced prostanoid formation and vasoconstriction in rabbit lungs. , 1986, The American review of respiratory disease.

[37]  R. Simon,et al.  OXIDANT ACTIVITY IN EXPIRED BREATH OF PATIENTS WITH ADULT RESPIRATORY DISTRESS SYNDROME , 1986, The Lancet.

[38]  A. Fishman.  Hypoxia on the pulmonary circulation. How and where it acts. , 1976, Circulation research.