ABSTRACT
Short-acting ß2-agonist bronchodilators are the most common medications used in treating chronic obstructive pulmonary disease (COPD). Genetic variants determining bronchodilator responsiveness (BDR) in COPD have not been identified. We performed a genome-wide association study (GWAS) of BDR in 5789 current or former smokers with COPD in one African-American and four white populations. BDR was defined as the quantitative spirometric response to inhaled ß2-agonists. We combined results in a meta-analysis. In the meta-analysis, single-nucleotide polymorphisms (SNPs) in the genes KCNK1 (P=2.02 × 10(-7)) and KCNJ2 (P=1.79 × 10(-7)) were the top associations with BDR. Among African Americans, SNPs in CDH13 were significantly associated with BDR (P=5.1 × 10(-9)). A nominal association with CDH13 was identified in a gene-based analysis in all subjects. We identified suggestive association with BDR among COPD subjects for variants near two potassium channel genes (KCNK1 and KCNJ2). SNPs in CDH13 were significantly associated with BDR in African Americans.The Pharmacogenomics Journal advance online publication, 27 October 2015; doi:10.1038/tpj.2015.65.
Subject(s)
Adrenergic beta-2 Receptor Agonists/therapeutic use , Bronchodilator Agents/therapeutic use , Lung/drug effects , Pharmacogenomic Variants/genetics , Pulmonary Disease, Chronic Obstructive/drug therapy , Pulmonary Disease, Chronic Obstructive/genetics , Black or African American/genetics , Aged , Cadherins/genetics , Europe , Female , Genome-Wide Association Study , Genotype , Humans , Lung/physiopathology , Male , Middle Aged , New Zealand , North America , Pharmacogenomic Testing , Phenotype , Potassium Channels, Inwardly Rectifying/genetics , Potassium Channels, Tandem Pore Domain/genetics , Pulmonary Disease, Chronic Obstructive/diagnosis , Pulmonary Disease, Chronic Obstructive/physiopathology , Risk Factors , Sarcoglycans/genetics , Severity of Illness Index , Spirometry , Treatment Outcome , White People/geneticsABSTRACT
BACKGROUND: The purpose was to define the differences between centrilobular (CLE) and panlobular emphysema (PLE) phenotypes in cigarette smokers with COPD by a combined qualitative-quantitative computed tomography (CT) analysis. METHODS: Chest CT scans of 116 cigarette smokers were visually scored by 22 chest radiologists and 29 pulmonologists in a single setting for the predominant emphysema phenotype (e.g. CLE or PLE) and automatically quantified for emphysema{% low attenuation area (LAA) ≤ -950 HU - %LAAinsp-950, gas trapping extent and bronchial metrics{wall area % for segmental (%WAsegm) and subsegmental (%WAsubsegm) bronchi}. These quantitative CT indexes were compared and related to FEV1, FEV1/FVC, and smoking history as stratified for emphysema phenotype. RESULTS: Although more frequent than CLE in GOLD stages 3 and 4 (p = 0.01), PLE was also scored in 38.2% of combined GOLD stages 1 and 2. PLE was positively associated with %LAAinsp-950 (OR = 1.18, 95% CI: 1.12 to 1.27, ß coefficient = 0.17, p = <0.0001) and negatively associated with pack-years of smoking (OR = 0.97, 95% CI: 0.95 to 0.99, ß coefficient = -0.02, p = 0.03). Both %WAsegm and %WAsubsegm were more strongly associated with FEV1% (R2 = 0.6 for both measures, p< 0.001) in CLE as compared to PLE (R2= 0.15, p = 0.02; R2 = 0.26, p< 0.001). CONCLUSIONS: PLE likely represents a more advanced phase of emphysema, which may also occur in earlier COPD stages and show different interplay with airway disease as compared to CLE.
ABSTRACT
Due to radiation-induced immune depression and development of pathologies such as cancer, there is increasing urgency to identify radiomitigators that are effective when administered after radiation exposure. The main goal of this study was to determine the radiomitigation capacity of MnTE-2-PyP[Mn(III) tetrakis (N-ethylpyridinium-2-yl) porphyrin], a superoxide dismutase (SOD) mimetic, and evaluate leukocyte parameters in spleen and blood. C57BL/6 mice were total-body exposed to 2 Gy γ-rays (Co-60), i.e., well below a lethal dose, followed by subcutaneous implantation of 5 × 10(5) RM-9 prostate tumor cells and initiation of MnTE-2-PyP treatment (day 0); interval between each procedure was 1-2 h. The drug was administered daily (12 times). Tumor progression was monitored and immunological analyses were performed on a subset per group on day 12. Animals treated with MnTE-2-PyP alone had significantly slower tumor growth compared to mice that did not receive the drug (P < 0.05), while radiation alone had no effect. Treatment of tumor-bearing mice with MnTE-2-PyP alone significantly increased spleen mass relative to body mass; the numbers of splenic white blood cells (WBC) and lymphocytes (B and T), as well as circulating WBC, granulocytes, and platelets, were high compared to one of more of the other groups (P < 0.05). The results show that MnTE-2-PyP slowed RM-9 tumor progression and up-regulated immune parameters, but mitigation of the effects of 2 Gy total-body irradiation were minimal.
Subject(s)
Antioxidants/administration & dosage , Metalloporphyrins/administration & dosage , Prostatic Neoplasms/radiotherapy , Radiation-Protective Agents/administration & dosage , Animals , Blood Cell Count , Male , Metalloporphyrins/chemistry , Mice , Mice, Inbred C57BL , Molecular Mimicry , Organ Size , Prostatic Neoplasms/immunology , Prostatic Neoplasms/pathology , Radiation Injuries, Experimental/prevention & control , Spleen/drug effects , Spleen/pathology , Spleen/radiation effects , Superoxide Dismutase/chemistry , Tumor Burden/drug effects , Tumor Burden/radiation effects , Whole-Body Irradiation , Xenograft Model Antitumor AssaysABSTRACT
This article investigates the suitability of local intensity distributions to analyze six emphysema classes in 342 CT scans obtained from 16 sites hosting scanners by 3 vendors and a total of 9 specific models in subjects with Chronic Obstructive Pulmonary Disease (COPD). We propose using kernel density estimation to deal with the inherent sparsity of local intensity histograms obtained from scarcely populated regions of interest. We validate our approach by leave-one-subject-out classification experiments and full-lung analyses. We compare our results with recently published LBP texture-based methodology. We demonstrate the efficacy of using intensity information alone in multi-scanner cohorts, which is a simpler, more intuitive approach.
ABSTRACT
Two primary chitinases have been identified in humans--acid mammalian chitinase (AMCase) and chitotriosidase (CHIT1). Mammalian chitinases have been observed to affect the host's immune response. The aim of this study was to test for association between genetic variation in the chitinases and phenotypes related to chronic obstructive pulmonary disease (COPD). Polymorphisms in the chitinase genes were selected based on previous associations with respiratory diseases. Polymorphisms that were associated with lung function level or rate of decline in the Lung Health Study (LHS) cohort were analyzed for association with COPD affection status in four other COPD case-control populations. Chitinase activity and protein levels were also related to genotypes. In the caucasian LHS population, the baseline forced expiratory volume in one second (FEV(1)) was significantly different between the AA and GG genotypic groups of the AMCase rs3818822 polymorphism. Subjects with the GG genotype had higher AMCase protein and chitinase activity compared with AA homozygotes. For CHIT1 rs2494303, a significant association was observed between rate of decline in FEV(1) and the different genotypes. In the African American LHS population, CHIT1 rs2494303 and AMCase G339T genotypes were associated with rate of decline in FEV(1). Although a significant effect of chitinase gene alleles was found on lung function level and decline in the LHS, we were unable to replicate the associations with COPD affection status in the other COPD study groups.
Subject(s)
Chitinases/genetics , Forced Expiratory Volume , Polymorphism, Single Nucleotide , Pulmonary Disease, Chronic Obstructive/genetics , Pulmonary Disease, Chronic Obstructive/physiopathology , Aged , Bronchoalveolar Lavage Fluid/chemistry , Case-Control Studies , Chitinases/metabolism , Female , Genetic Variation , Genotype , Humans , Lung/metabolism , Lung/physiopathology , Male , Middle Aged , Phenotype , Pulmonary Disease, Chronic Obstructive/enzymology , Respiratory Physiological Phenomena , SmokingABSTRACT
PURPOSE: The purpose of this study was to evaluate the efficacy of the antioxidant Mn (III) tetrakis (N-ethylpyridinium-2-yl) porphyrin (MnTE-2-PyP) in protecting ocular tissue and retinal microvasculature from radiation damage. MATERIALS AND METHODS: 75 rats were treated with Mn TE-2-PyP at 2.5 micro g/injection into one eye an hour before proton irradiation. The radiation was delivered in a single fraction to total doses of 8 Gray (Gy) or 28 Gy; Rats were sacrificed 3 days and 3, 6, 9, and 12 months thereafter for histology and quantification of photoreceptor cell populations and retinal capillary changes. RESULTS: By 6 months following radiation, there was significant loss of retinal outer and inner nuclear layers in eyes receiving radiation only (8 and 28 Gy) (p < 0.05) compared to their controls and to the eyes of rats treated with radiation plus metalloporphyrin. Retinal microvessel length density decreased significantly 6 months following 28 Gy (p < 0.05) compared to their controls and to MnTE-2-PyP treated rats. By 12 months following irradiation, irradiated eyes showed extensive damage to the photoreceptor layer, whereas the eyes of animals receiving radiation plus MnTE-2-PyP showed almost no morphological damage. MnTE-2-PyP treatment also suppressed radiation-induced apoptosis in our study. CONCLUSIONS: These results demonstrated that MnTE-2-PyP protected both photoreceptors and retinal capillaries from radiation damage, suggesting that this metalloporphyrin antioxidant is effective in regulating the damage induced by proton radiation.
Subject(s)
Antioxidants/therapeutic use , Metalloporphyrins/therapeutic use , Radiation Injuries, Experimental/prevention & control , Radiation-Protective Agents/therapeutic use , Retinal Diseases/prevention & control , Animals , Apoptosis , Caspase 3/metabolism , Cataract/classification , Cataract/etiology , Cataract/prevention & control , Fluorescent Antibody Technique, Indirect , Lens, Crystalline/radiation effects , Male , Photoreceptor Cells, Vertebrate/enzymology , Photoreceptor Cells, Vertebrate/pathology , Photoreceptor Cells, Vertebrate/radiation effects , Protons , Radiation Dosage , Radiation Injuries, Experimental/etiology , Radiation Injuries, Experimental/pathology , Rats , Rats, Sprague-Dawley , Retina/radiation effects , Retinal Diseases/etiology , Retinal Diseases/pathology , Retinal Vessels/pathology , Retinal Vessels/radiation effects , Treatment OutcomeABSTRACT
OBJECTIVE: Excess reactive oxygen species and oxidative damage have been associated with the pathogenesis of osteoarthritis (OA). Extracellular superoxide dismutase (EC-SOD or SOD3) scavenges superoxide is the major catalytic antioxidant in joint fluid and is decreased in OA cartilage. We studied human joint fluid samples to test whether there is an association between OA and EC-SOD or other low molecular antioxidants in the joint fluid. METHODS: Joint fluid samples were obtained from 28 subjects with severe OA undergoing arthrocentesis or knee joint replacement and compared to joint fluid from 12 subjects undergoing knee arthroscopy for chronic knee pain, meniscal tears or anterior cruciate ligament reconstruction. EC-SOD protein was assayed by enzyme-linked immunosorbent assay (ELISA). Ascorbate and urate were measured with high performance liquid chromatography (HPLC) and total nitrates by the Greiss reaction. Glutathione (GSH) and oxidized glutathione were measured using a colorimetric method. Interleukin-6 (IL-6) and transforming growth factor-beta (TGF-beta) were both measured with ELISA. RESULTS: Human joint fluid contains significant amounts of the extracellular, catalytic antioxidant EC-SOD. Joint fluid from OA subjects is characterized by significantly decreased EC-SOD levels and significant decreases in GSH, and ascorbate compared to the reference group of knee joints with pain or subacute injury but macroscopically intact cartilage. GSH and ascorbate show only an age effect with no effect from disease state on regression modeling. Urate is present in joint fluid but does not show a significant difference between groups. IL-6 and TGF-beta both show non-significant trends to increases in the arthritic subjects. There was no correlation of EC-SOD levels with IL-6 as a marker of inflammation in either the comparison group or the OA group. CONCLUSIONS: EC-SOD, the major scavenger of reactive oxygen species (ROS) in extracellular spaces and fluids, is decreased in late stage OA joint fluid compared to fluid from injured/painful joints with intact cartilage. Injured joints may be able to increase or maintain secretion of EC-SOD but it appears that late stage OA joints fail to do so in spite of increased oxidative stress seen in the disease. Associated age related declines in GSH and ascorbate might also contribute to the development of severe OA. The net effect of these changes in joint fluid antioxidants is likely to accelerate the damaging oxidant effects on extracellular matrix stability in cartilage tissue.
Subject(s)
Cartilage, Articular/metabolism , Free Radical Scavengers/metabolism , Osteoarthritis, Knee/metabolism , Superoxide Dismutase/metabolism , Synovial Fluid/metabolism , Age Factors , Cartilage, Articular/physiopathology , Enzyme-Linked Immunosorbent Assay , Extracellular Matrix/metabolism , Female , Free Radical Scavengers/pharmacology , Glutathione , Humans , Interleukin-6/metabolism , Knee Joint/metabolism , Knee Joint/physiopathology , Male , Middle Aged , Nitrates , Nitrites , Osteoarthritis, Knee/physiopathology , Pain , Reactive Oxygen Species/adverse effects , Reactive Oxygen Species/metabolism , Superoxide Dismutase/pharmacology , Transforming Growth Factor beta/metabolism , Uric Acid/analysisABSTRACT
Tumor necrosis factor (TNF)-alpha, a cytokine present in inflammed lungs, is known to mediate some of the adverse effects of ozone and inhaled particles. The authors evaluated transgenic mice with constitutive pulmonary expression of TNF-alpha under transcriptional regulation of the surfactant protein-C promoter as an animal model of biological susceptibility to air pollutants. To simulate a repeated, episodic exposure to air pollutants, wild-type and TNF mice inhaled air or a mixture of ozone (0.4 ppm) and urban particles (EHC-93, 4.8 mg/m3) for 4 h, once per week, for 12 consecutive weeks and were sacrificed 20 h after last exposure. TNF mice exhibited chronic lung inflammation with septal thickening, alveolar enlargement, and elevated protein and cellularity in bronchoalveolar lavage fluid (genotype main effect, p < .001). Repeated exposure to pollutants did not result in measurable inflammatory changes in wild-type mice and did not exacerbate the inflammation in TNF mice. The pollutants decreased recovery of alveolar macrophages in tavage fluid of both wild-type and TNF mice (exposure main effect, p < .001). Exacerbation of the rate of protein nitration reactions specifically in the lungs of TNF mice was revealed by the high ratio of 3-nitrotyrosine to L-DOPA after exposure to the air pollutants (Genotype x Exposure factor interaction, p = .014). Serum creatine kinase-MM isoform increased in TNF mice exposed to pollutants (Genotype X Exposure factor interaction, p = .043). The marked pollutant-related nitration in the lungs of the TNF mice reveals basic differences in free radical generation and scavenging in the inflamed lungs in response to pollutants. Furthermore, elevation of circulating creatine kinase-MM isoform specifically in TNF mice exposed to pollutants suggests systemic adverse impacts from lung inflammatory mediators, possibly on muscles and the cardiovascular system.
Subject(s)
Air Pollutants/toxicity , Inhalation Exposure/adverse effects , Lung/drug effects , Pneumonia/chemically induced , Tumor Necrosis Factor-alpha/metabolism , Administration, Inhalation , Animals , Bronchoalveolar Lavage Fluid/chemistry , Bronchoalveolar Lavage Fluid/cytology , Chronic Disease , Creatine Kinase/blood , Creatine Kinase, MM Form , Disease Models, Animal , Endothelins/genetics , Endothelins/metabolism , Female , Isoenzymes/blood , Lung/metabolism , Lung/pathology , Macrophages, Alveolar/drug effects , Macrophages, Alveolar/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Pneumonia/metabolism , Pneumonia/pathology , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Toxicity Tests , Tumor Necrosis Factor-alpha/geneticsABSTRACT
Oxidant stress is a key mechanism for smoking-induced chronic obstructive pulmonary disease (COPD). Smoking has been shown to upregulate several antioxidant enzymes, with potential effects on the prevention of the disease and/or its progression. Superoxide dismutases (SOD)s are the only enzymes capable of consuming superoxide radicals. The purpose of the present study was to investigate SODs in the lungs of nonsmokers, smokers and COPD patients. Manganese superoxide dismutase (MnSOD), copper zinc SOD (CuZnSOD), and extracellular SOD (ECSOD), were investigated by immunohistochemistry in the airways of 13 nonsmokers, 20 smokers and 22 COPD patients with mild-to-moderate disease. Lung tissue homogenates of three nonsmokers and four smokers were used for Western blot and enzyme activity analysis. The expression of MnSOD was higher in the central bronchial epithelium of smokers with COPD and in the alveolar epithelium of smokers without or with COPD than innonsmokers. Lung MnSOD immunoreactivity, evaluated by Western blotting and specific activity, were 33% and 51% higher, respectively, in smokers than in nonsmokers. No major changes could be observed in lung CuZnSOD or ECSOD immunoreactivities. Manganese superoxide dismutase is elevated in the alveolar epithelium of cigarette smokers, probably due to the increased oxidant burden in smokers' lungs.
Subject(s)
Lung/enzymology , Smoking/metabolism , Superoxide Dismutase/analysis , Aged , Aldehydes/analysis , Blotting, Western , Bronchi/enzymology , Epithelium/enzymology , Female , Humans , Immunohistochemistry , Male , Pulmonary Alveoli/enzymology , Pulmonary Disease, Chronic Obstructive/enzymology , Smoking/adverse effectsABSTRACT
This article characterised the role of oxidative stress in mediating pathological reactions in the lung and the unique antioxidant defences that the lung possesses. The impact of redox balance in regulating inflammatory and immune reactions were discussed, and the impact of enhancing lung antioxidant capacity in animal models of asthma and chronic obstructive pulmonary disease were characterised.
Subject(s)
Cell Death/physiology , Cytokines/biosynthesis , Oxidation-Reduction , Oxidative Stress , Pneumonia/pathology , Animals , Cells, Cultured , Cytokines/analysis , Humans , Inflammation Mediators/metabolism , Lipid Peroxidation , Lung/cytology , Lung/pathology , Pneumonia/enzymology , Pneumonia/physiopathology , Prognosis , Respiratory System/pathology , Respiratory System/physiopathology , Sensitivity and Specificity , Superoxide Dismutase/metabolismABSTRACT
Oxidative stress plays an important role in the development of fibrotic responses in the lung. However, it is not clear whether inhibiting oxidative stress with antioxidants can attenuate fibrotic processes in the lung. The objective of these studies was to test whether the catalytic antioxidant porphyrin manganese (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) could protect mice against bleomycin-induced lung fibrosis. A 10 mg/kg intraperitoneal dose of MnTBAP was established as safe and had a serum and lung half-life of 9.5 h in mice. Based on this data, four groups of mice were given one dose of bleomycin (3.2 U/kg, intratracheal) or saline and MnTBAP (5 mg/kg, intraperitoneal) or saline twice daily for 14 d. Lung fibrosis was assessed by measuring (1) lung hydroxyproline content as an index of collagen accumulation, (2) airway dysfunction by whole body plethysmography, and (3) histopathology. Bleomycin produced a 20% loss in body weight that was only 10% in the bleomycin/MnTBAP group. Bleomycin produced a twofold increase in hydroxyproline content that was decreased 23% by MnTBAP. Bleomycin produced a twofold increase in airway dysfunction that was also attenuated 30% by MnTBAP. Histopathologic analysis of the lungs of mice treated with bleomycin demonstrated a severe fibrotic response that was attenuated 28% by MnTBAP. Future studies on the oxidant mechanisms that MnTBAP is affecting in this bleomycin model of lung fibrosis may shed light on potential new therapeutic approaches for treating interstitial lung diseases.
Subject(s)
Antioxidants/pharmacology , Bleomycin/antagonists & inhibitors , Bleomycin/toxicity , Metalloporphyrins/pharmacology , Pulmonary Fibrosis/chemically induced , Pulmonary Fibrosis/prevention & control , Animals , Antioxidants/pharmacokinetics , Antioxidants/toxicity , Collagen/metabolism , Hydroxyproline/metabolism , Male , Metalloporphyrins/pharmacokinetics , Metalloporphyrins/toxicity , Mice , Mice, Inbred BALB C , Oxidative Stress , Pulmonary Fibrosis/metabolism , Pulmonary Fibrosis/pathologyABSTRACT
Reperfusion of the lung after hemorrhage generates free radicals such as superoxide (O(2)(.)) that may injure the lung; however, the relative importance of intracellular versus extracellular free radicals is unclear. The superoxide dismutases (SOD) are the primary enzymatic method to reduce superoxide. We examined whether lung-specific overexpression of extracellular superoxide dismutase (EC-SOD) would attenuate hemorrhage-induced lung injury. Wild-type mice and mice overexpressing the human EC-SOD gene with a lung-specific promoter were hemorrhaged by removing 30% of blood volume. After hemorrhage, the lung wet to dry weight ratios increased from 5.4 +/- 0.11 in unmanipulated control mice to 6.3 +/- 0.16 in wild-type mice, but to only 5.60 +/- 0.17 in the EC-SOD transgenic mice (p < 0.05 compared with hemorrhaged wild-type). Hemorrhage-induced lipid peroxidation, as assessed by lung F(2) isoprostanes, was lower in the EC-SOD transgenic mice (3.4 +/- 0.3 microg/lung) compared with wild-type mice (1.9 +/- 0.2 microg/lung; p < 0.05). Compared with wild-type, EC-SOD transgenic mice had attenuated the hemorrhage-induced increase in both pulmonary nuclear factor kappa B (NK-kappaB) activation (relative absorbance 1.1 +/- 0.2 for EC-SOD transgenic versus 2.5 +/- 0.1 for wild-type; p < 0.05) and myeloperoxidase activity (5.1 +/- 0.87 units/g for EC-SOD transgenic versus 11.3 +/- 1.8 units/g for wild-type; p < 0.01). Thus, overexpression of pulmonary EC-SOD in the mouse lung attenuates lung injury after hemorrhage.
Subject(s)
Free Radical Scavengers/therapeutic use , Hemorrhage/complications , Lung Diseases/drug therapy , Superoxide Dismutase/therapeutic use , Animals , Lung Diseases/etiology , MiceABSTRACT
Reactive oxygen species contribute to ischemic brain injury. This study examined whether the porphyrin catalytic antioxidant manganese (III) meso-tetrakis (N-ethylpyridinium-2-yl)porphyrin (MnTE-2-PyP(5+)) reduces oxidative stress and improves outcome from experimental cerebral ischemia. Rats that were subjected to 90 min focal ischemia and 7 d recovery were given MnTE-2-PyP(5+) (or vehicle) intracerebroventricularly 60 min before ischemia, or 5 or 90 min or 6 or 12 hr after reperfusion. Biomarkers of brain oxidative stress were measured at 4 hr after postischemic treatment (5 min or 6 hr). MnTE-2-PyP(5+), given 60 min before ischemia, improved neurologic scores and reduced total infarct size by 70%. MnTE-2-PyP(5+), given 5 or 90 min after reperfusion, reduced infarct size by 70-77% and had no effect on temperature. MnTE-2-PyP(5+) treatment 6 hr after ischemia reduced total infarct volume by 54% (vehicle, 131 +/- 60 mm(3); MnTE-2-PyP(5+), 300 ng, 60 +/- 68 mm(3)). Protection was observed in both cortex and caudoputamen, and neurologic scores were improved. No MnTE-2-PyP(5+) effect was observed if it was given 12 hr after ischemia. MnTE-2-PyP(5+) prevented mitochondrial aconitase inactivation and reduced 8-hydroxy-2'-deoxyguanosine formation when it was given 5 min or 6 hr after ischemia. In mice, MnTE-2-PyP(5+) reduced infarct size and improved neurologic scores when it was given intravenously 5 min after ischemia. There was no effect of 150 or 300 ng of MnTE-2-PyP(5+) pretreatment on selective neuronal necrosis resulting from 10 min forebrain ischemia and 5 d recovery in rats. Administration of a metalloporphyrin catalytic antioxidant had marked neuroprotective effects against focal ischemic insults when it was given up to 6 hr after ischemia. This was associated with decreased postischemic superoxide-mediated oxidative stress.
Subject(s)
Antioxidants/administration & dosage , Brain Ischemia/drug therapy , Cerebral Infarction/prevention & control , Metalloporphyrins/administration & dosage , Neuroprotective Agents/administration & dosage , Aconitate Hydratase/metabolism , Animals , Antioxidants/chemistry , Brain/blood supply , Brain/drug effects , Brain/enzymology , Brain/pathology , Brain Ischemia/etiology , Catalysis , Cerebral Infarction/etiology , DNA/metabolism , Disease Models, Animal , Dose-Response Relationship, Drug , Drug Administration Schedule , Fumarate Hydratase/metabolism , Infarction, Middle Cerebral Artery/complications , Injections, Intravenous , Injections, Intraventricular , Male , Metalloporphyrins/chemistry , Mice , Mice, Inbred C57BL , Mitochondria/enzymology , Necrosis , Neuroprotective Agents/chemistry , Oxidative Stress/drug effects , Rats , Rats, Wistar , Treatment OutcomeABSTRACT
Extracellular superoxide dismutase (EC-SOD) controls the availability of extracellular superoxide and appears to play a role in controlling intercellular signaling. In this role EC-SOD can have potent effects on neurobehavioral function. In previous studies, we have found that either over- or under-expression of EC-SOD in mice significantly impairs spatial learning on the radial-arm maze. In the current study, the neurobehavioral nature of the EC-SOD role in cognitive function was determined. EC-SOD overexpression altered the relationship between both learning and memory with motivational state. Mice were tested in the radial-arm maze under a high motivational state (22-24 hours of food restriction) or a low motivational state (4-6 hours of food restriction). Under a high motivational state, the EC-SOD overexpressing mice were able to learn in the radial-arm maze, albeit at a slightly lower rate than wild-type controls. This contrasts with the failure to learn by EC-SOD overexpressing mice in our previous study conducted with the low motivational state. The change in motivational state did not significantly alter the learning rate of controls. Similarly, during postacquisition memory phase of testing, the EC-SOD overexpressing mice were significantly worse than controls when tested in a low motivational state but not under a high motivation state. As with learning, motivational state did not significantly affect memory performance in controls. This study shows that mice with EC-SOD overexpression are not incapable of learning and memory in the radial-arm maze, but that the mechanisms which allow control animals to perform this task well under low motivational states are deficient in the mice with EC-SOD overexpression.
Subject(s)
Appetitive Behavior/physiology , Maze Learning/physiology , Mental Recall/physiology , Motivation , Superoxide Dismutase/genetics , Animals , Extracellular Space/enzymology , Female , Food Deprivation , Gene Expression Regulation, Enzymologic/physiology , MiceABSTRACT
OBJECTIVE: The aim of this study was to determine placental localization and activity of extracellular superoxide dismutase, a nitric oxide modulator, during early gestation and to correlate these characteristics with fetal vascular development. STUDY DESIGN: First-trimester (n = 10) and second-trimester (n = 10) villi were obtained at elective pregnancy termination. Extracellular superoxide dismutase was localized by means of an immunoperoxidase method. Activity was measured by determining the inhibition of cytochrome c reduction at pH 10 and messenger ribonucleic acid expression by in situ hybridization. RESULTS: Extracellular superoxide dismutase was intracellular within villous trophoblasts until 17 weeks' gestation, when it relocated to the villous extracellular matrix. Activities were similar between first- and second-trimester villi. In situ hybridization confirmed extracellular superoxide dismutase messenger ribonucleic acid within trophoblasts throughout gestation. CONCLUSION: Extracellular superoxide dismutase is produced by trophoblasts early in pregnancy, but it remains intracellular until 17 weeks' gestation, which may be related to fetal vascular development.
Subject(s)
Placenta/enzymology , Superoxide Dismutase/analysis , Blotting, Western , Cytochrome c Group/metabolism , Extracellular Space/enzymology , Female , Gestational Age , Humans , Hydrogen-Ion Concentration , Immunoenzyme Techniques , In Situ Hybridization , Keratins/analysis , Pregnancy , RNA, Messenger/analysis , Trophoblasts/enzymologyABSTRACT
Extracellular superoxide dismutase (EC-SOD) is the major isozyme of SOD in arteries, but is also abundant in lungs. In particular, mouse lungs contain large amounts of EC-SOD compared to lungs in other mammals. This suggests that EC-SOD may have an amplified function in the mouse lung. This study describes the purification and characterization of mouse EC-SOD as well as its localization in mouse lung. Mouse EC-SOD exists primarily as a homotetramer composed of a pair of dimers linked through disulfide bonds present in the heparin-binding domains of each subunit. In addition, mouse EC-SOD can exist in active multimeric forms. We developed and utilized a polyclonal antibody to mouse EC-SOD to immunolocalize EC-SOD in mouse lung. EC-SOD labeling is strongest in the matrix of vessels, airways, and alveolar septa. This localization suggests that EC-SOD may have important functions in pulmonary biology, perhaps in the modulation of nitric oxide-dependent responses.
Subject(s)
Lung/enzymology , Superoxide Dismutase/isolation & purification , Amino Acid Sequence , Animals , Chromatography, Gel , Electrophoresis, Polyacrylamide Gel , Immunohistochemistry , Mice , Molecular Sequence Data , Superoxide Dismutase/chemistry , Superoxide Dismutase/metabolismABSTRACT
Transgenic mice, which exhibit a fivefold increase in brain parenchymal extracellular superoxide dismutase (EC-SOD) activity, were used to investigate the role of EC-SOD in global ischemic brain injury. Halothane-anesthetized normothermic wild-type (n = 22) and transgenic (n = 20) mice underwent 10 min of near-complete forebrain ischemia induced by bilateral carotid artery occlusion and systemic hypotension (mean arterial pressure = 30 mm Hg). After 3 days of recovery, the brains were histologically examined. Other mice underwent autoradiographic determination of regional CBF 10 min prior to, during, and 30 min after forebrain ischemia. Histologic injury in the cortex and caudoputamen was minimal in both groups. The percentage of dead hippocampal CA1 neurons was reduced in the EC-SOD transgenic group (wild type = 44 +/- 28%; EC-SOD transgenic = 23 +/- 21%, mean +/- SD, P = 0.015). CBF was similar between groups prior to ischemia. The intraischemic blood flow was severely reduced in forebrain structures and was similar between groups. Blood flow at 30 min postischemia had recovered to 50-60% of baseline values in both groups. These results indicate that EC-SOD can play an important role in defining the magnitude of selective neuronal necrosis resulting from near-complete forebrain ischemia. This implicates involvement of extracellular superoxide anions in the pathologic response to global cerebral ischemia.
Subject(s)
Brain Ischemia/metabolism , Prosencephalon/metabolism , Superoxide Dismutase/metabolism , Animals , Brain Ischemia/pathology , Male , Mice , Mice, Inbred C3H , Mice, Inbred C57BL , Mice, Transgenic , Prosencephalon/blood supply , Prosencephalon/pathologyABSTRACT
Emphysema is commonly defined as enlargement of airspaces distal to terminal bronchioles accompanied by destruction of alveolar walls, but without obvious fibrosis. Morphometric techniques were used to correlate changes in components of the alveolar septa surrounding enlarged airspaces in human emphysema with the mean linear intercept (Lm) of those airspaces. Alveolar and capillary surface density decreased with increased Lm, but the ratio of these surface densities to each other remained close to normal for mild to moderate increases in Lm. This suggests that the decreased gas exchange observed in emphysema is initiated by a total loss of septa and not by selective pathological changes of the microvasculature. Increases in septal wall thickness directly correlated with increases in Lm. For the mild to moderate emphysema lesions included in this study, an increase of 100% in Lm correlated with a 130% increase in the relative volume of the alveolar septal interstitium. Significant increases occurred in both elastin (0.14 to 0.56 microm(3)/microm(2) basement membrane [BM]) and collagen (0.49 to 1. 63 microm(3)/microm(2) BM). The increase in elastin and collagen raises the possibility of a remodeling process in the connective matrix in alveolar walls. Whether or not the new connective tissue represents a disordered, nonfunctional regional response needs to be determined.