Effects of S-nitrosation and cross-linking of hemoglobin on hypoxic pulmonary vasoconstriction in isolated rat lungs.
Free hemoglobin (Hb) and red blood cells augment hypoxic pulmonary vasoconstriction (HPV) by scavenging nitric oxide (NO). S-nitrosation of Hb (SNO-Hb) may confer vasodilatory properties by allowing release of NO during deoxygenation and/or by interaction with small-molecular weight thiols. Likewise, cross-linking of free Hb may limit its vasoconstrictive effect by preventing abluminal movement of the molecule. We compared the effects of free SNO-Hb and Hb intramolecularly cross-linked at the beta-cysteine 93 residue [Bis(maleidophenyl)-polyethylene glycol2000HbA (Bis-Mal-PEGHb)] to those of free oxyHb on pulmonary artery pressure (PAP), HPV, and exhaled NO (eNO) in isolated, perfused rat lungs. Ventilation of lungs with anoxic gas for 5 minutes reduced perfusate PO2 to 11+/-1.0 Torr. Addition of SNO-Hb or Bis-Mal-PEGHb (100 micromol/L) to buffer perfusate increased normoxic PAP and augmented HPV in similar magnitude as free oxyHb, but had no effect on eNO. Addition of the allosteric modulator inositol hexaphosphate to increase Hb P50 and the thiol glutathione (GSH) to allow removal of NO from Hb via transnitrosation to the perfusate did not reduce augmentation of HPV by SNO-Hb or increase eNO. GSH resulted in an approximately 50% reduction in perfusate [S-nitrosothiol], in association with an increase in perfusate [metHb]. Free SNO-Hb is a net NO scavenger and pulmonary vasoconstrictor in this model, although thiol-mediated release of NO from SNO-Hb does occur. However, release of NO from SNO-Hb was not influenced by deoxygenation-mediated allosteric changes in Hb across a broad range of oxyHb saturation. Cross-linking of Hb does not limit its pulmonary vasoconstrictor effects.
D000818:Animals; D015687:Cell Hypoxia; D003432:Cross-Linking Reagents / Q000494:pharmacology; D016166:Free Radical Scavengers / Q000737:chemistry / Q000494:pharmacology*; D005978:Glutathione / Q000494:pharmacology; D006454:Hemoglobins / Q000737:chemistry / Q000494:pharmacology*; D008168:Lung / Q000098:blood supply / Q000187:drug effects / Q000502:physiology*; D008301:Maleimides / Q000494:pharmacology; D009569:Nitric Oxide / Q000032:analysis; D015538:Nitrosation; D009924:Organ Culture Techniques; D010833:Phytic Acid / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D011651:Pulmonary Artery / Q000502:physiology*; D011669:Pulmonary Wedge Pressure; D051381:Rats; D026422:S-Nitrosoglutathione / Q000494:pharmacology; D014661:Vasoconstriction / Q000187:drug effects; D014662:Vasoconstrictor Agents / Q000737:chemistry / Q000494:pharmacology*
D003432:Cross-Linking Reagents; D016166:Free Radical Scavengers; D006454:Hemoglobins; D008301:Maleimides; C103574:S-nitrosohemoglobin; D014662:Vasoconstrictor Agents; C405685:bis(maleidophenyl)PEG2000; D009569:Nitric Oxide; D011092:Polyethylene Glycols; D026422:S-Nitrosoglutathione; D010833:Phytic Acid; D005978:Glutathione