Dithionite increases radical formation and decreases vasoconstriction in the lung: Evidence that dithionite does not mimic alveolar hypoxia

Stephen L. Archer, Václav Hampl, Daniel P. Nelson, Erika Sidney, Douglas A Peterson, Edward K Weir

Research output: Contribution to journalArticle

48 Scopus citations

Abstract

Dithionite is a powerful reducing agent used to deoxygenate hemoglobin and create anaerobic conditions in vitro. Recently, dithionite has been used as a convenient means of creating 'hypoxia' in experiments studying the O2 sensor in the pulmonary circulation and carotid body. We evaluated the hypothesis that hypoxia created by hypoxic ventilation and that created by dithionite have different effects on the pulmonary circulation. In vitro, dithionite (10-5 to 10-3 mol/L), added to oxygenated Krebs' solution, rapidly created superoxide anion in a dose-dependent manner. Dithionite consumed O2 in parallel with the generation of superoxide radical, with both processes peaking within seconds. Anoxia was sustained only if resupply of O2 was prevented. In isolated rat lungs (whether perfused with autologous blood or Krebs' solution), hypoxic ventilation alone lowered perfusate PO2 from ≃140 to 40 mm Hg and decreased lung levels of activated oxygen species (AOS), measured by luminol-enhanced chemiluminescence, before the onset of hypoxic pulmonary vasoconstriction. Constrictor responses to angiotensin II and KCl were not impaired by intermittent hypoxic challenges, and lung weight did not increase. In contrast, dithionite impaired constrictor responses of the Krebs' solution-perfused lungs to all vasoconstrictors tested and increased lung weight. When given as a bolus (5 x 10-3 mol/L) into the pulmonary artery during normoxic ventilation, dithionite caused no vasoconstriction and only briefly lowered PO2 (because of constant resupply of O2 from the alveoli). When superimposed on hypoxic ventilation, dithionite further lowered PO2 from ≃40 to ≃0 mm Hg and caused additional constriction. Unlike hypoxic ventilation, dithionite increased AOS production. Antioxidant enzymes diminished dithionite-induced radical production and diminished the loss of vascular reactivity and lung edema. In conclusion, unlike hypoxic ventilation, dithionite causes edema and loss of vascular reactivity in the lung by generating superoxide anion and hydrogen peroxide. Hypoxia elicited by dithionite is not equivalent to authentic hypoxia because of the obligatory associated generation of AOS. Dithionite usage should not be substituted for authentic hypoxia in studies of O2 sensing.

Original languageEnglish (US)
Pages (from-to)174-181
Number of pages8
JournalCirculation Research
Volume77
Issue number1
DOIs
StatePublished - Jul 1995

Keywords

  • chemiluminescence
  • dithionite
  • hypoxia
  • oxygen radicals
  • oxygen sensor

Fingerprint Dive into the research topics of 'Dithionite increases radical formation and decreases vasoconstriction in the lung: Evidence that dithionite does not mimic alveolar hypoxia'. Together they form a unique fingerprint.

  • Cite this