1-3-dimethylthiourea and Lung-Diseases

Dimethylthiourea (DMTU) is an effective scavenger of reactive oxygen metabolites. This property has been successfully exploited, experimentally, in the protection of cells and tissues against oxidative damage. In this study, however, we have observed that levels of nonprotein sulfhydryls (NPSH) in hamster lung slices were markedly decreased by incubation with 10 or 40 mM DMTU. These changes were associated with morphological signs of injury, increased levels of oxidised glutathione (GSSG), and an increased activity of the pentose phosphate pathway (PPP), suggesting that the loss of NPSH was due to their oxidation. Incubation with 40 mM, but not 10 mM DMTU, also resulted in a decreased ability to oxidise [6-14C]glucose or to synthesise proteins, suggesting that at the high concentration, DMTU may cause functional impairment of the tissue. Furthermore, the ability of the slices to accumulate putrescine decreased after incubation with the oxidative toxins paraquat (PQ), tert-butyl hydroperoxide (t-BOOH) or hydrogen peroxide (H2O2) and was further decreased by co-incubation with DMTU. Putrescine uptake, a function specific to the alveolar type I and II epithelial cells, was not affected by incubation with DMTU alone. DMTU did not exacerbate the effect of the nonoxidative toxin iodoacetamide (IAA) on putrescine uptake but it did affect markers of general cell damage or dysfunction. We suggest, therefore, that the toxicity of oxidants toward lung tissue is potentiated in alveolar epithelial cells by DMTU.

Topics: Animals; Cricetinae; Drug Synergism; Female; Free Radicals; Hydrogen Peroxide; Iodoacetamide; Lung; Lung Diseases; Male; Mesocricetus; Microscopy, Electron; Oxidants; Oxidation-Reduction; Paraquat; Peroxides; Putrescine; tert-Butylhydroperoxide; Thiourea

Using lucigenin-enhanced chemiluminescence, isolated rat lungs perfused with physiological salt-Ficoll solution were studied to test whether phorbol myristate acetate (PMA)-induced lung injury was mediated by reactive oxygen species (ROS). PMA (0.03 micrograms ml-1) caused small but significant increases in lung ROS levels and pulmonary arterial perfusion pressure (Ppa) but did not induce lung oedema. PMA (0.15 micrograms ml-1) induced lung oedema with large increases in ROS production and Ppa. Superoxide dismutase (SOD) inhibited the increases in ROS, Ppa, and lung oedema. Catalase and dimethylthiourea inhibited lung oedema but did not attenuate the increases in ROS and Ppa entirely. Indomethacin attenuated lung oedema partially but did not inhibit the increases in ROS and Ppa. These data indicate that PMA-induced lung injury is dependent on PMA concentration and ROS are responsible for such lung injury. Thromboxane plays a minor role for PMA-induced lung injury. The different effects of oxygen radical scavengers suggest that different radical species contribute to the increased pulmonary vascular response and lung injury.

Topics: Acridines; Animals; Catalase; Indomethacin; Luminescent Measurements; Lung; Lung Diseases; Male; Pressure; Pulmonary Artery; Pulmonary Edema; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Superoxide Dismutase; Tetradecanoylphorbol Acetate; Thiourea

In this study, we proposed that oxygen free radicals participate in the acute pulmonary injury that follows limb ischemia/reperfusion. Using an established model of hind limb ischemia, reproducible lung injury occurred after reperfusion. Lung microvascular permeability was measured with 125I-BSA and increased two-fold after 30 minutes of reperfusion. Pulmonary injury was blocked with DMSO, DMTU, allopurinol, indomethacin, and SOD plus catalase. The degree of pulmonary neutrophil sequestration as assessed by tissue myeloperoxidase activity was significantly diminished in animals pretreated with antioxidants. Pretreatment with indomethacin did not attenuate the neutrophil sequestration within the pulmonary parenchyma. These data suggest that increased lung microvascular permeability and neutrophil accumulation occur following hind limb ischemia/reperfusion. Therapeutic interventions with oxygen radical inhibitors blocked this process, while the prostaglandin inhibitor, indomethacin, only reduced lung permeability.

Topics: Allopurinol; Animals; Capillary Permeability; Catalase; Dimethyl Sulfoxide; Free Radicals; Hindlimb; Indomethacin; Lung; Lung Diseases; Neutrophils; Peroxidase; Rats; Rats, Inbred Strains; Reperfusion Injury; Superoxide Dismutase; Thiourea

Injection of phorbol 12-myristate 13-acetate (PMA) into polymorphonuclear leukocyte (PMN)-depleted, PMN cytoplast-repleted New Zealand White rabbits caused the development of acute lung injury in vivo. PMN cytoplasts are nucleus- and granule-free vesicles of cytoplasm capable of releasing toxic O2 radicals but incapable of releasing granule enzymes. PMN cytoplasts when activated by PMA reduced 66 +/- 12.7 nmol of cytochrome c compared with 2.6 +/- 0.7 nmol in their resting state and did not release a significant quantity of granule enzymes (P greater than 0.05). Injection of PMA into New Zealand White rabbits caused a significant decrease (P less than 0.05) in the number of circulating cytoplasts. Increases in lung weight-to-body weight ratios in PMA-treated rabbits (9.8 +/- 0.5 X 10(-3] compared with saline-treated rabbits (5.3 +/- 0.2 X 10(-3] were also noted. Levels of angiotensin-converting enzyme in lung lavage as well as the change in alveolar-arterial O2 ratio correlated with the numbers of cytoplasts in lung lavage (P = 0.001, r = 0.84 and P = 0.0166, r = 0.73, respectively). Albumin in lung lavage increased to 1,700 +/- 186 mg/ml in PMA-treated rabbits from 60 +/- 30 mg/ml in saline-treated rabbits. These changes were attenuated by pretreatment of rabbits with dimethylthiourea (DMTU). In vitro, cytoplasts were able to mediate increases in endothelial monolayer permeability. This was evidenced by increases in fractional transit of albumin across endothelial monolayers when treated with PMA-activated cytoplasts (0.08 +/- 0.01 to 0.28 +/- 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Bronchoalveolar Lavage Fluid; Cell Membrane Permeability; Chemotaxis, Leukocyte; Endothelium; Humans; Lung; Lung Diseases; Neutrophils; Peptidyl-Dipeptidase A; Pulmonary Edema; Rabbits; Superoxides; Tetradecanoylphorbol Acetate; Thiourea

The effects of hydroxyl radical inhibition on an experimental model of normobaric pulmonary oxygen toxicity have been studied. The metal ion chelator, desferrioxamine (which inhibits hydroxyl-radical generation) or the hydroxyl-radical scavenger, dimethylthiourea (DMTU), were administered in an attempt to block hydroxyl-radical-mediated tissue injury. Lung injury was monitored in Sprague-Dawley rats by examining lung histology and bronchoalveolar lavage and by assessing pulmonary capillary permeability using the 125I-albumin lung permeability index and the lung weight:body weight ratio. Control animals had lung permeability indices 0.183 +/- 0.005 and lung weight to body weight ratio of 4.50 +/- 0.10 (all as mean +/- SEM). With increased duration of exposure to hyperoxia, there was a progressive increase in pulmonary inflammation, with thickening of alveolar membranes and atelectasis and a progressive increase in lung permeability indices (0.434 +/- 0.088 at 24 hrs; 0.954 +/- 0.165 at 48 hrs; and 1.55 +/- 0.214 at 60 hrs); and lung weight to body weight ratio (5.28 +/- 0.11 at 24 hrs; 6.54 +/- 0.23 at 48 hrs; and 8.91 +/- 0.51 at 60 hrs). Treatment with desferrioxamine provided significant protection from lung injury after 24 hrs of hyperoxia (eg., lung permeability indices 0.250 +/- 0.018; lung weight to body weight ratio 4.68 +/- 0.14, both p less than 0.025; cf. 24-hr hyperoxia controls) but no reduction in pulmonary injury was observed after 48 and 60 hrs of hyperoxia exposure.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Capillary Permeability; Deferoxamine; Hydroxides; Hydroxyl Radical; Lung; Lung Diseases; Male; Organ Size; Oxygen; Rats; Rats, Inbred Strains; Thiourea

Lung injury induced by phorbol myristate acetate (PMA) is closely associated with toxic oxidants released from activated granulocytes. But the major toxic oxidant causing lung damage is not really known. We have, therefore, conducted investigations using various oxygen radical scavengers. The intravenous administration of dimethylthiourea (DMTU), a potent hydroxyl radical scavenger, or of superoxide dismutase (SOD), a superoxide anion scavenger, plus catalase, a hydrogen peroxide scavenger, to rabbits intravenously injected with PMA prevented biochemical data and cellularity indicative of lung damage in lung lavages. Morphologically, the lungs of PMA-injected rabbits revealed mild interstitial edema, aggregates of granulocytes within the interstitial capillaries, and the increase of granulocytes in alveolar spaces. Furthermore, there was direct morphologic evidence of pulmonary endothelial cell disruption. In rabbits treated with DMTU or SOD plus catalase, there was no evidence of destructive changes in the lungs. SOD-treated rabbits did not show evidence of protection from PMA-induced lung injury. Only a little protection was provided by catalase treatment. Moreover, in the ultracytochemical study for examination of hydrogen peroxide (H2O2) generation, the number of H2O2-generated granulocytes remarkably decreased in lung lavages of catalase-treated rabbits, but destructive changes were observed in the lungs. In contrast, though the number of H2O2-generated granulocyte was not decreased in lung lavages of DMTU-treated rabbits, treatment with DMTU could afford protection from lung injury. These data indicate that the hydroxyl radical, a toxic oxidant derived from stimulated granulocytes, is deeply involved in the pathogenesis of PMA-induced lung injury.

Topics: Animals; Body Weight; Bronchoalveolar Lavage Fluid; Catalase; Cell Count; Free Radicals; Granulocytes; Histocytochemistry; Hydrogen Peroxide; Hydroxides; Hydroxyl Radical; Lung; Lung Diseases; Male; Organ Size; Rabbits; Superoxide Dismutase; Superoxides; Tetradecanoylphorbol Acetate; Thiourea

Previous studies from our laboratory have demonstrated that thermal injury to the skin of rats is associated with the production of oxygen radicals by complement-activated blood neutrophils, resulting in acute lung injury as demonstrated by increases in lung vascular permeability and morphological evidence of vascular endothelial cell damage, interstitial edema, and alveolar hemorrhage. In the present study, the analysis of sera from thermally injured rats reveals an isoenzyme profile for lactate dehydrogenase (LDH;EC 1.1.1.27) that is compatible with origin from lung. The appearance of LDH-4 isoenzyme in serum of thermally injured rats correlates linearly with indices of lung damage, supporting the results of previous studies suggesting that thermal trauma to the skin can cause oxygen radical production by complement-activated blood neutrophils with resultant acute microvascular injury in the lung interstitium. Furthermore, interventions that protect from oxidant-mediated lung injury (catalase, scavengers of hydroxyl radical, iron chelators or neutrophil depletion) result in significant reductions in serum levels of the LDH-4 isoenzyme following thermal injury to the skin. Thus, measurements of LDH isoenzyme patterns in serum appear to be useful in monitoring tissue damage such as oxygen radical-mediated acute lung injury.

Topics: Animals; Burns; Deferoxamine; Free Radicals; Isoenzymes; L-Lactate Dehydrogenase; Lung; Lung Diseases; Lung Injury; Neutrophils; Oxidants, Photochemical; Oxygen; Rats; Rats, Inbred Strains; Skin; Thiourea