ABSTRACT
BACKGROUND AND OBJECTIVES: In a proportion of patients, dementia with Lewy bodies (DLB) is associated with Alzheimer disease (AD) copathology, which is linked to accelerated cognitive decline and more extensive cortical atrophy. The objective was to evaluate the relationship between a biomarker of AD copathology, plasma tau phosphorylated at residue 181 (ptau181), and the treatment effects of the p38α kinase inhibitor neflamapimod, which targets the cholinergic degenerative process in DLB. METHODS: The AscenD-LB study was a phase 2a, randomized (1:1), 16-week, placebo-controlled clinical trial of neflamapimod in DLB, the main results of which have been published. After the study was completed (i.e., post hoc), pretreatment plasma ptau181 levels were determined and participants were grouped based on a cutoff for AD pathology of 2.2 pg/mL (established in a separate cohort to identify AD from healthy controls). Clinical outcomes for the comparison of placebo with neflamapimod 40 mg three times daily (TID; the higher and more clinically active of 2 doses studied) were analyzed using mixed models for repeated measures within each subgroup (baseline plasma ptau181 < and ≥2.2 pg/mL). RESULTS: Pretreatment plasma ptau181 levels were determined in eighty-five participants with mild-to-moderate DLB receiving cholinesterase inhibitors, with 45 participants below and 40 above the 2.2 pg/mL cutoff at baseline. In the 16-week treatment period, in the comparison of placebo with neflamapimod 40 mg TID, for all end points evaluated, improvements with neflamapimod treatment were greater in participants below the cutoff, compared with those above the cutoff. In addition, participants below the ptau181 cutoff at baseline showed significant improvement over placebo in an attention composite measure (+0.42, 95% CI 0.07-0.78, p = 0.023, d = 0.78), the Clinical Dementia Rating Scale Sum of Boxes (-0.60, 95% CI -1.04 to -0.06, p = 0.031, d = 0.70), the Timed Up and Go test (-3.1 seconds, 95% CI -4.7 to -1.6, p < 0.001, d = 0.74), and International Shopping List Test-Recognition (+1.4, 95% CI 0.2-2.5, p = 0.024, d = 1.00). DISCUSSION: Exclusion of patients with elevated plasma ptau181, potentially through excluding patients with extensive cortical neurodegeneration, enriches for a patient with DLB population that is more responsive to neflamapimod. More generally, plasma biomarkers of AD copathology at study entry should be considered as stratification variables in DLB clinical trials. TRIAL REGISTRATION INFORMATION: NCT04001517 at ClinicalTrials.gov.
Subject(s)
Alzheimer Disease , Lewy Body Disease , Humans , Alzheimer Disease/pathology , Biomarkers , Cholinesterase Inhibitors/therapeutic use , Lewy Body Disease/drug therapy , Lewy Body Disease/complications , Postural Balance , Protein Kinase Inhibitors/therapeutic use , Time and Motion StudiesABSTRACT
Opa1 participates in inner mitochondrial membrane fusion and cristae morphogenesis. Here, we show that muscle-specific Opa1 ablation causes reduced muscle fiber size, dysfunctional mitochondria, enhanced Fgf21, and muscle inflammation characterized by NF-κB activation, and enhanced expression of pro-inflammatory genes. Chronic sodium salicylate treatment ameliorated muscle alterations and reduced the muscle expression of Fgf21. Muscle inflammation was an early event during the progression of the disease and occurred before macrophage infiltration, indicating that it is a primary response to Opa1 deficiency. Moreover, Opa1 repression in muscle cells also resulted in NF-κB activation and inflammation in the absence of necrosis and/or apoptosis, thereby revealing that the activation is a cell-autonomous process and independent of cell death. The effects of Opa1 deficiency on the expression NF-κB target genes and inflammation were absent upon mitochondrial DNA depletion. Under Opa1 deficiency, blockage or repression of TLR9 prevented NF-κB activation and inflammation. Taken together, our results reveal that Opa1 deficiency in muscle causes initial mitochondrial alterations that lead to TLR9 activation, and inflammation, which contributes to enhanced Fgf21 expression and to growth impairment.
Subject(s)
DNA, Mitochondrial/genetics , GTP Phosphohydrolases/physiology , Inflammation/etiology , Muscle, Skeletal/pathology , Muscular Diseases/etiology , Toll-Like Receptor 9/metabolism , Animals , Apoptosis , Cells, Cultured , Cytokines/metabolism , Female , Inflammation/metabolism , Inflammation/pathology , Male , Mice, Knockout , Muscle, Skeletal/immunology , Muscular Diseases/metabolism , Muscular Diseases/pathology , Necrosis , Regeneration , Toll-Like Receptor 9/geneticsABSTRACT
PURPOSE: To investigate the outcome of suppression of the renin angiotensin system using captopril combined with an antioxidant (Eukarion [EUK]-207) for mitigation of radiation-induced lung damage in rats. METHODS AND MATERIALS: The thoracic cavity of female Sprague-Dawley rats was irradiated with a single dose of 11 Gy. Treatment with captopril at a dose of 40 mg/kg/d in drinking water and EUK-207 given by subcutaneous injection (8 mg/kg daily) was started 1 week after irradiation (PI) and continuing until 14 weeks PI. Breathing rate was monitored until the rats were killed at 32 weeks PI, when lung fibrosis was assessed by lung hydroxyproline content. Lung levels of the cytokine transforming growth factor-ß1 and macrophage activation were analyzed by immunohistochemistry. Oxidative DNA damage was assessed by 8-hydroxy-2-deoxyguanosine levels, and lipid peroxidation was measured by a T-BARS assay. RESULTS: The increase in breathing rate in the irradiated rats was significantly reduced by the drug treatments. The drug treatment also significantly decreased the hydroxyproline content, 8-hydroxy-2-deoxyguanosine and malondialdehyde levels, and levels of activated macrophages and the cytokine transforming growth factor-ß1 at 32 weeks. Almost complete mitigation of these radiation effects was observed by combining captopril and EUK-207. CONCLUSION: Captopril and EUK-207 can provide mitigation of radiation-induced lung damage out to at least 32 weeks PI after treatment given 1-14 weeks PI. Overall the combination of captopril and EUK-207 was more effective than the individual drugs used alone.
Subject(s)
Captopril/pharmacology , Lung/radiation effects , Organometallic Compounds/pharmacology , Radiation Injuries, Experimental/drug therapy , Radiation Pneumonitis/drug therapy , Radiation-Protective Agents/pharmacology , Renin-Angiotensin System/drug effects , 8-Hydroxy-2'-Deoxyguanosine , Animals , Captopril/administration & dosage , DNA Damage , Deoxyguanosine/analogs & derivatives , Deoxyguanosine/analysis , Drug Therapy, Combination/methods , Female , Lipid Peroxidation , Lung/chemistry , Organometallic Compounds/administration & dosage , Radiation-Protective Agents/administration & dosage , Rats , Rats, Sprague-Dawley , Respiratory Rate/drug effects , Thiobarbituric Acid Reactive Substances/analysis , Transforming Growth Factor beta1/analysisABSTRACT
The 1918 influenza pandemic caused over 40 million deaths worldwide, with 675,000 deaths in the United States alone. Studies in several experimental animal models showed that 1918 influenza virus infection resulted in severe lung pathology associated with dysregulated immune and cell death responses. To determine if reactive oxygen species produced by host inflammatory responses play a central role in promoting severity of lung pathology, we treated 1918 influenza virus-infected mice with the catalytic catalase/superoxide dismutase mimetic, salen-manganese complex EUK-207 beginning 3 days postinfection. Postexposure treatment of mice infected with a lethal dose of the 1918 influenza virus with EUK-207 resulted in significantly increased survival and reduced lung pathology without a reduction in viral titers. In vitro studies also showed that EUK-207 treatment did not affect 1918 influenza viral replication. Immunohistochemical analysis showed a reduction in the detection of the apoptosis marker cleaved caspase-3 and the oxidative stress marker 8-oxo-2'-deoxyguanosine in lungs of EUK-207-treated animals compared to vehicle controls. High-throughput sequencing and RNA expression microarray analysis revealed that treatment resulted in decreased expression of inflammatory response genes and increased lung metabolic and repair responses. These results directly demonstrate that 1918 influenza virus infection leads to an immunopathogenic immune response with excessive inflammatory and cell death responses that can be limited by treatment with the catalytic antioxidant EUK-207.
Subject(s)
Free Radical Scavengers/pharmacology , Influenza A Virus, H1N1 Subtype/physiology , Influenza Pandemic, 1918-1919 , Organometallic Compounds/pharmacology , Orthomyxoviridae Infections/drug therapy , Reactive Oxygen Species/antagonists & inhibitors , 8-Hydroxy-2'-Deoxyguanosine , Animals , Biomarkers/metabolism , Caspase 3/genetics , Caspase 3/metabolism , DNA Repair , Deoxyguanosine/analogs & derivatives , Deoxyguanosine/metabolism , Dogs , Female , Gene Expression , Humans , Inflammation/drug therapy , Inflammation/metabolism , Inflammation/mortality , Inflammation/virology , Influenza A Virus, H1N1 Subtype/pathogenicity , Lung/drug effects , Lung/metabolism , Lung/pathology , Madin Darby Canine Kidney Cells , Mice , Mice, Inbred BALB C , Orthomyxoviridae Infections/metabolism , Orthomyxoviridae Infections/mortality , Orthomyxoviridae Infections/virology , Reactive Oxygen Species/metabolism , Survival Analysis , Viral Load , Virus ReplicationABSTRACT
PURPOSE: Radioprotection and mitigation effects of the antioxidants, Eukarion (EUK)-207, curcumin, and the curcumin analogs D12 and D68, on radiation-induced DNA damage or lipid peroxidation in murine skin were investigated. These antioxidants were studied because they have been previously reported to protect or mitigate against radiation-induced skin reactions. METHODS: DNA damage was assessed using two different assays. A cytokinesis-blocked micronucleus (MN) assay was performed on primary skin fibroblasts harvested from the skin of C3H/HeJ male mice 1 day, 1 week and 4 weeks after 5 Gy or 10 Gy irradiation. Local skin or whole body irradiation (100 kVp X-rays or caesium (Cs)-137 γ-rays respectively) was performed. DNA damage was further quantified in keratinocytes by immunofluorescence staining of γ-histone 2AX (γ-H2AX) foci in formalin-fixed skin harvested 1 hour or 1 day post-whole body irradiation. Radiation-induced lipid peroxidation in the skin was investigated at the same time points as the MN assay by measuring malondialdehyde (MDA) with a Thiobarbituric acid reactive substances (TBARS) assay. RESULTS: None of the studied antioxidants showed significant mitigation of skin DNA damage induced by local irradiation. However, when EUK-207 or curcumin were delivered before irradiation they provided some protection against DNA damage. In contrast, all the studied antioxidants demonstrated significant mitigating and protecting effects on radiation-induced lipid peroxidation at one or more of the three time points after local skin irradiation. CONCLUSION: Our results show no evidence for mitigation of DNA damage by the antioxidants studied in contrast to mitigation of lipid peroxidation. Since these agents have been reported to mitigate skin reactions following irradiation, the data suggest that changes in lipid peroxidation levels in skin may reflect developing skin reactions better than residual post-irradiation DNA damage in skin cells. Further direct comparison studies are required to confirm this inference from the data.
Subject(s)
DNA Damage , Free Radical Scavengers/pharmacology , Lipid Peroxidation/drug effects , Lipid Peroxidation/radiation effects , Radiation-Protective Agents/pharmacology , Skin/drug effects , Skin/radiation effects , Animals , Curcumin/chemistry , Curcumin/pharmacology , Free Radical Scavengers/chemistry , Male , Malondialdehyde/metabolism , Mice , Micronucleus Tests , Radiation-Protective Agents/chemistry , Skin/metabolismABSTRACT
Cranial irradiation with (56)Fe, a form of space radiation, causes hippocampus-dependent cognitive changes. (56)Fe irradiation also increases reactive oxygen species (ROS) levels, which may contribute to these changes. Therefore, we investigated the effects of the antioxidant alpha lipoic acid (ALA) on cognition following sham-irradiation and irradiation. Male mice were irradiated (brain only) with (56)Fe (3 Gy) or sham-irradiated at 6-9 months of age. Half of the mice remained fed a regular chow and the other half of the mice were fed a caloric-matched diet containing ALA starting two-weeks prior to irradiation and throughout cognitive testing. Following cognitive testing, levels of 3-nitrotyrosine (3NT), a marker of oxidative protein stress, and levels of microtubule-associated protein (MAP-2), a dendritic protein important for cognition, were assessed using immunohistochemistry and confocal microscopy. ALA prevented radiation-induced impairments in spatial memory retention in the hippocampal and cortical dependent water maze probe trials following reversal learning. However, in sham-irradiated mice, ALA treatment impaired cortical-dependent novel object recognition and amygdala-dependent cued fear conditioning. There was a trend towards lower 3NT levels in irradiated mice receiving a diet containing ALA than irradiated mice receiving a regular diet. In the hippocampal dentate gyrus of mice on regular diet, irradiated mice had higher levels of MAP-2 immunoreactivity than sham-irradiated mice. Thus, ALA might have differential effects on the brain under normal physiological conditions and those involving environmental challenges such as cranial irradiation.
Subject(s)
Cranial Irradiation/psychology , Iron/toxicity , Memory Disorders/diet therapy , Memory/drug effects , Memory/radiation effects , Radiation Injuries, Experimental/diet therapy , Thioctic Acid/pharmacology , Animals , Antioxidants/pharmacology , Antioxidants/therapeutic use , Brain/drug effects , Brain/metabolism , Brain/radiation effects , Conditioning, Psychological/drug effects , Conditioning, Psychological/radiation effects , Cranial Irradiation/methods , Dentate Gyrus/drug effects , Dentate Gyrus/metabolism , Dentate Gyrus/radiation effects , Male , Maze Learning/drug effects , Maze Learning/radiation effects , Memory Disorders/chemically induced , Mice , Mice, Inbred C57BL , Microtubule-Associated Proteins/metabolism , Radiation Injuries, Experimental/chemically induced , Thioctic Acid/therapeutic use , Tyrosine/analogs & derivatives , Tyrosine/metabolismABSTRACT
In the event of a radionuclear attack or nuclear accident, the skin would be the first barrier exposed to radiation, though skin injury can progress over days to years following exposure. Chronic oxidative stress has been implicated as being a potential contributor to the progression of delayed radiation-induced injury to skin and other organs. To examine the causative role of oxidative stress in delayed radiation-induced skin injury, including impaired wound healing, we tested a synthetic superoxide dismutase (SOD)/catalase mimetic, EUK-207, in a rat model of combined skin irradiation and wound injury. Administered systemically, beginning 48 hours after irradiation, EUK-207 mitigated radiation dermatitis, suppressed indicators of tissue oxidative stress, and enhanced wound healing. Evaluation of gene expression in irradiated skin at 30 days after exposure revealed a significant upregulation of several key genes involved in detoxication of reactive oxygen and nitrogen species. This gene expression pattern was primarily reversed by EUK-207 therapy. These results demonstrate that oxidative stress has a critical role in the progression of radiation-induced skin injury, and that the injury can be mitigated by appropriate antioxidant compounds administered 48 hours after exposure.
Subject(s)
Organometallic Compounds/pharmacology , Oxidative Stress/drug effects , Radiation Injuries, Experimental/drug therapy , Radiodermatitis/drug therapy , Wound Healing/drug effects , Animals , Antioxidants/pharmacology , Catalase/metabolism , Disease Models, Animal , Gene Expression/drug effects , Gene Expression/physiology , Male , Molecular Mimicry/physiology , Oxidative Stress/genetics , Radiation Injuries, Experimental/metabolism , Radiation Injuries, Experimental/pathology , Radiodermatitis/metabolism , Radiodermatitis/pathology , Rats , Rats, Inbred Strains , Skin/blood supply , Skin/pathology , Skin/radiation effects , Superoxide Dismutase/metabolism , Wound Healing/physiologyABSTRACT
In the event of a radiological accident or terrorist attack, whole- or partial-body exposure can injure the lungs. To simulate such an incident, we used a single fraction of total-body irradiation (TBI) or whole-thoracic irradiation to induce pneumonitis or pulmonary fibrosis, respectively, in a rat model. The superoxide dismutase and catalase mimetic EUK-207 was given by subcutaneous injection (20 mg/kg/day, 5 days per week, once daily) starting at 7 days after irradiation and stopping before pneumonitis developed. After TBI, morbidity and the increase in breathing rates associated with pneumonitis were significantly improved in rats treated with EUK-207 compared to rats receiving irradiation alone. At 42 days after TBI (the peak of pneumonitis) changes in vascular end points including pulmonary hemodynamics ex vivo and relative arterial density in lungs were also mitigated by EUK-207. At 7 months after whole-thoracic irradiation, EUK-207 reduced synthesis of collagen as assessed by the Sircol collagen assay and Masson's trichrome staining. Our results demonstrate promise for EUK-207 as a mitigator of radiation pneumonitis and fibrosis. We also demonstrate for the first time mitigation of multiple vascular injuries in the irradiated lung in vivo by EUK-207.
Subject(s)
Biomimetic Materials/administration & dosage , Organometallic Compounds/administration & dosage , Radiation Pneumonitis/drug therapy , Radiation-Protective Agents/administration & dosage , Animals , Catalase/chemistry , Lung/drug effects , Lung/pathology , Lung/radiation effects , Radiation Pneumonitis/pathology , Rats , Superoxide Dismutase/chemistry , Thorax/drug effects , Thorax/pathology , Thorax/radiation effects , Whole-Body IrradiationABSTRACT
BACKGROUND: Oxidative stress and mitochondrial dysfunction are central mediators of cardiac dysfunction after ischemia/reperfusion. ATP binding cassette mitochondrial erythroid (ABC-me; ABCB10; mABC2) is a mitochondrial transporter highly induced during erythroid differentiation and predominantly expressed in bone marrow, liver, and heart. Until now, ABC-me function in heart was unknown. Several lines of evidence demonstrate that the yeast ortholog of ABC-me protects against increased oxidative stress. Therefore, ABC-me is a potential modulator of the outcome of ischemia/reperfusion in the heart. METHODS AND RESULTS: Mice harboring 1 functional allele of ABC-me (ABC-me(+/-)) were generated by replacing ABC-me exons 2 and 3 with a neomycin resistance cassette. Cardiac function was assessed with Langendorff perfusion and echocardiography. Under basal conditions, ABC-me(+/-) mice had normal heart structure, hemodynamic function, mitochondrial respiration, and oxidative status. However, after ischemia/reperfusion, the recovery of hemodynamic function was reduced by 50% in ABC-me(+/-) hearts as a result of impairments in both systolic and diastolic function. This reduction was associated with impaired mitochondrial bioenergetic function and with oxidative damage to both mitochondrial lipids and sarcoplasmic reticulum calcium ATPase after reperfusion. Treatment of ABC-me(+/-) hearts with the superoxide dismutase/catalase mimetic EUK-207 prevented oxidative damage to mitochondria and sarcoplasmic reticulum calcium ATPase and restored mitochondrial and cardiac function to wild-type levels after reperfusion. CONCLUSIONS: Inactivation of 1 allele of ABC-me increases the susceptibility to oxidative stress induced by ischemia/reperfusion, leading to increased oxidative damage to mitochondria and sarcoplasmic reticulum calcium ATPase and to impaired functional recovery. Thus, ABC-me is a novel gene that determines the ability to tolerate cardiac ischemia/reperfusion.
Subject(s)
ATP-Binding Cassette Transporters/genetics , Mitochondria/physiology , Myocardial Reperfusion Injury/genetics , Myocardial Reperfusion Injury/metabolism , Oxidative Stress/genetics , Recovery of Function/genetics , ATP-Binding Cassette Transporters/metabolism , Animals , Cardiac Volume/physiology , Catalase/metabolism , Female , Genetic Predisposition to Disease/genetics , Male , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Mitochondria/drug effects , Mutagenesis, Insertional , Myocardial Contraction/drug effects , Myocardial Contraction/physiology , Myocardial Reperfusion Injury/drug therapy , Organometallic Compounds/pharmacology , Oxidative Stress/drug effects , Superoxide Dismutase/metabolism , Ventricular Pressure/physiologyABSTRACT
PURPOSE: We examined the effects of genistein and/or Eukarion (EUK)-207 on radiation-induced lung damage and investigated whether treatment for 0-14 weeks (wks) post-irradiation (PI) would mitigate late lung injury. MATERIALS AND METHODS: The lungs of female Sprague-Dawley (SD) rats were irradiated with 10 Gy. EUK-207 was delivered by infusion and genistein was delivered as a dietary supplement starting immediately after irradiation (post irradiation [PI]) and continuing until 14 wks PI. Rats were sacrificed at 0, 4, 8, 14 and 28 wks PI. Breathing rate was monitored and lung fibrosis assessed by lung hydroxyproline content at 28 wks. DNA damage was assessed by micronucleus (MN) assay and 8-hydroxy-2-deoxyguanosine (8-OHdG) levels. The expression of the cytokines Interleukin (IL)-1α, IL-1ß, IL-6, Tumor necrosis factor (TNF)-α and Transforming growth factor (TGF)-ß1, and macrophage activation were analyzed by immunohistochemistry. RESULTS: Increases in breathing rate observed in the irradiated rats were significantly reduced by both drug treatments during the pneumonitis phase and the later fibrosis phase. The drug treatments decreased micronuclei (MN) formation from 4-14 wks but by 28 wks the MN levels had increased again. The 8-OHdG levels were lower in the drug treated animals at all time points. Hydroxyproline content and levels of activated macrophages were decreased at 28 wks in all drug treated rats. The treatments had limited effects on the expression of the cytokines. CONCLUSION: Genistein and EUK-207 can provide partial mitigation of radiation-induced lung damage out to at least 28 wks PI even after cessation of treatment at 14 wks PI.
Subject(s)
Genistein/administration & dosage , Organometallic Compounds/administration & dosage , Radiation Pneumonitis/drug therapy , Radiation Pneumonitis/physiopathology , Radiation-Protective Agents/administration & dosage , Respiratory Mechanics/drug effects , Respiratory Mechanics/radiation effects , Animals , Female , Rats , Rats, Sprague-Dawley , Treatment OutcomeABSTRACT
Salen Mn complexes, including EUK-134, EUK-189 and a newer cyclized analog EUK-207, are synthetic SOD/catalase mimetics that have beneficial effects in many models of oxidative stress. As oxidative stress is implicated in some forms of delayed radiation injury, we are investigating whether these compounds can mitigate injury to normal tissues caused by ionizing radiation. This review describes some of this research, focusing on several tissues of therapeutic interest, namely kidney, lung, skin, and oral mucosa. These studies have demonstrated suppression of delayed radiation injury in animals treated with EUK-189 and/or EUK-207. While an antioxidant mechanism of action is postulated, it is likely that the mechanisms of radiation mitigation by these compounds in vivo are complex and may differ in the various target tissues. Indicators of oxidative stress are increased in lung and skin radiation injury models, and suppressed by salen Mn complexes. The role of oxidative stress in the renal injury model is unclear, though EUK-207 does mitigate. In certain experimental models, salen Mn complexes have shown "mito-protective" properties, that is, attenuating mitochondrial injury. Consistent with this, EUK-134 suppresses effects of ionizing radiation on mitochondrial function in rat astrocyte cultures. In summary, salen Mn complexes could be useful to mitigate delayed radiation injury to normal tissues following radiation therapy, accidental exposure, or radiological terrorism. Optimization of their mode of delivery and other key pharmaceutical properties, and increasing understanding of their mechanism(s) of action as radiation mitigators, are key issues for future study.
Subject(s)
Ethylenediamines/pharmacology , Ethylenediamines/therapeutic use , Organometallic Compounds/pharmacology , Organometallic Compounds/therapeutic use , Radiation Injuries/drug therapy , Radiation Injuries/prevention & control , Animals , Humans , Oxidative Stress/drug effects , Radiation Injuries/metabolism , Radiation Injuries, Experimental/drug therapy , Radiation Injuries, Experimental/metabolism , Radiation Injuries, Experimental/prevention & control , Superoxide Dismutase/pharmacology , Superoxide Dismutase/therapeutic useSubject(s)
Biomimetic Materials/chemistry , Superoxide Dismutase/chemistry , Superoxide Dismutase/metabolism , Animals , Biomimetic Materials/pharmacology , Humans , Manganese/metabolism , Organometallic Compounds/chemistry , Organometallic Compounds/pharmacology , Polyamines/chemistry , Polyamines/pharmacology , Porphyrins/chemistry , Porphyrins/pharmacologyABSTRACT
Numerous in vitro and in vivo studies have shown that the endothelial cells of the microvasculature of the lung and kidney are damaged by exposure to ionizing radiation, and this sustained endothelial cell injury is involved in the early and late radiation effects observed in these tissues. It is well accepted that ionizing radiation causes the generation of reactive oxygen species during exposure that results in damage to DNA and cellular organelles. It is more controversial, however, whether additional biochemical events or long-lived radicals occur and persist postirradiation that amplify and initiate new forms of cellular damage. Two families of Eukarion (EUK) compounds have been synthesized that possess superoxide dismutase (SOD), catalase and peroxidase activities. The Mn porphyrins are available orally whereas the salen Mn complexes are administered by injection. In the present study we have examined the ability of these SOD/catalase mimetics to prevent apoptosis of endothelial cells when administered 1 h postirradiation (mitigation). A range of salen Mn complex (EUK-189 and EUK-207) and Mn porphyrins (EUK-418, -423, -425, -450, -451, -452, -453) were used to treat endothelial cells 1 h after the cells received 2-20 Gy ionizing radiation in vitro. Two lead compounds, EUK-207 at a dose of 30 microM and EUK-451 at a dose of 10 microM, exhibited low toxicity and mitigated radiation-induced apoptosis. Future animal studies will test whether these compounds protect when administered after radiation exposure as would be done after a radiological accident or a terrorism event.
Subject(s)
Apoptosis/drug effects , Apoptosis/radiation effects , Biomimetic Materials/pharmacology , Catalase/metabolism , Endothelial Cells/drug effects , Endothelial Cells/radiation effects , Superoxide Dismutase/metabolism , Animals , Biomimetic Materials/administration & dosage , Biomimetic Materials/chemical synthesis , Biomimetic Materials/toxicity , Caspases/biosynthesis , Caspases/metabolism , Cell Line , Cell Nucleus/drug effects , Cell Nucleus/metabolism , Cell Nucleus/radiation effects , Dose-Response Relationship, Radiation , Endothelial Cells/cytology , Endothelial Cells/metabolism , L-Lactate Dehydrogenase/metabolism , Necrosis/chemically induced , Organometallic Compounds/administration & dosage , Organometallic Compounds/chemical synthesis , Organometallic Compounds/pharmacology , Organometallic Compounds/toxicity , Toxicity TestsABSTRACT
Superoxide dismutase/catalase mimetics, such as salen Mn complexes and certain metalloporphyrins, catalytically neutralize reactive oxygen and nitrogen species, which have been implicated in the pathogenesis of many serious diseases. Both classes of mimetic are protective in animal models of oxidative stress. However, only AEOL11207 and EUK-418, two uncharged Mn porphyrins, have been shown to be orally bioavailable. In this study, EUK-418 and several new analogs (the EUK-400 series) were synthesized and shown to exhibit superoxide dismutase, catalase, and peroxidase activities in vitro. Some also protected PC12 cells against staurosporine-induced cell death. All EUK-400 compounds were stable in simulated gastric fluid, and most were substantially more lipophilic than the salen Mn complexes EUK-189 and EUK-207, which lack oral activity. Pharmacokinetics studies demonstrate the presence of all EUK-400 series compounds in the plasma of rats after oral administration. These EUK-400 series compounds are potential oral therapeutic agents for cellular damage caused by oxidative stress.
Subject(s)
Catalase/metabolism , Manganese/metabolism , Metalloporphyrins/administration & dosage , Metalloporphyrins/metabolism , Superoxide Dismutase/metabolism , Administration, Oral , Animals , Biocatalysis , Biological Availability , Biomimetic Materials/administration & dosage , Biomimetic Materials/metabolism , Biomimetic Materials/pharmacokinetics , Biomimetic Materials/pharmacology , Cell Death/drug effects , Cell Line, Tumor , Metalloporphyrins/pharmacokinetics , Metalloporphyrins/pharmacology , PC12 Cells , Rats , Staurosporine/pharmacologyABSTRACT
Animal models, and human postmortem studies, of prion disease have demonstrated the presence of heightened oxidative stress in the brain, with additional findings supporting the likelihood that the normal isoform of prion protein directly contributes to neuronal antioxidant defences. Although such data are consistent with the postulate that oxidative stress plays a salient pathogenic role in prion disease, it remains possible that oxidative damage represents a secondary or relatively less important phenomenon in neurons already rendered dysfunctional from other primary insults. To provide further insights into the relative pathogenic importance of oxidative stress, we employed a potent manganese-superoxide dismutase/catalase mimetic, EUK-189, as a therapeutic in our mouse model of human prion disease. A significant but relatively modest prolongation of survival in EUK-189-treated mice was observed, which correlated with reductions in oxidative, especially nitrative, damage to proteins when compared to untreated disease controls. Lesion profiling also revealed reductions in spongiform change in specific brain regions of terminally sick EUK-189-treated mice. Our results are consistent with heightened oxidative stress playing a pathogenic role in prion disease but underscore the need for more biologically potent and, most likely, broader spectrum antioxidant treatments if more successful amelioration is to be achieved.
Subject(s)
Antioxidants/therapeutic use , Brain/drug effects , Catalase , Organometallic Compounds/therapeutic use , Prion Diseases/drug therapy , Salicylates/therapeutic use , Superoxide Dismutase , Animals , Biomimetics , Blotting, Western , Brain/pathology , Disease Models, Animal , Humans , Immunohistochemistry , Mice , Prion Diseases/pathology , Prions/biosynthesis , Prions/drug effects , Protein Carbonylation/drug effectsABSTRACT
Extensive epidemiological data in humans and studies in animal models of Parkinson's disease (PD) suggest that sporadic forms of the disorder are not strictly genetic in nature but most likely because of combined environmental exposures over the period of the life-span coupled with increased genetic susceptibilities. Environmental paraquat and neonatal iron exposure have both been separately suggested as potential risk factors for sporadic forms of the disease. In this study, we demonstrate that combined environmental exposure to these two agents results in accelerated age-related degeneration of nigrostriatal dopaminergic neurons. Furthermore, pretreatment with the synthetic superoxide dismutase/catalase mimetic, EUK-189, significantly attenuated neuronal death mediated by combined paraquat and iron treatment. These findings support the notion that environmental PD risk factors may act synergistically to produce neurodegeneration associated with the disorder and that iron and paraquat may act via common oxidative stress-mediated mechanisms.
Subject(s)
Aging/metabolism , Environmental Exposure/adverse effects , Iron/toxicity , Nerve Degeneration/etiology , Paraquat/toxicity , Parkinson Disease/etiology , Aging/pathology , Animals , Cell Line , Cells, Cultured , Drug Synergism , Herbicides/toxicity , Male , Mice , Mice, Inbred C57BL , Nerve Degeneration/metabolism , Nerve Degeneration/physiopathology , Neurons/drug effects , Neurons/metabolism , Neurons/pathology , Neuroprotective Agents/pharmacology , Organometallic Compounds/pharmacology , Parkinson Disease/metabolism , Parkinson Disease/physiopathology , Rats , Salicylates/pharmacology , Substantia Nigra/drug effects , Substantia Nigra/pathology , Substantia Nigra/physiopathology , Superoxide Dismutase/metabolismABSTRACT
One postulated mechanism for the reduction in stress tolerance with aging is a decline in the regulation of stress-responsive genes, such as inducible heat shock protein 72 (HSP70). Increased levels of oxidative stress are also associated with aging, but it is unclear what impact a prooxidant environment might have on HSP70 gene expression. This study utilized a superoxide dismutase/catalase mimetic (Eukarion-189) to evaluate the impact of a change in redox environment on age-related HSP70 responses to a physiologically relevant heat challenge. Results demonstrate that liver HSP70 mRNA and protein levels are reduced in old compared with young rats at selected time points over a 48-h recovery period following a heat-stress protocol. While chronic systemic administration of Eukarion-189 suppressed hyperthermia-induced liver HSP70 mRNA expression in both age groups, HSP70 protein accumulation was blunted in old rats but not in their young counterparts. These data suggest that a decline in HSP70 mRNA levels may be responsible for the reduction in HSP70 protein observed in old animals after heat stress. Furthermore, improvements in redox status were associated with reduced HSP70 mRNA levels in both young and old rats, but differential effects were manifested on protein expression, suggesting that HSP70 induction is differentially regulated with aging. These findings highlight the integrated mechanisms of stress protein regulation in eukaryotic organisms responding to environmental stress, which likely involve interactions between a wide range of cellular signals.
Subject(s)
Aging/metabolism , Antioxidants/pharmacology , Biomimetic Materials/pharmacology , Fever/metabolism , HSP70 Heat-Shock Proteins/metabolism , Liver/drug effects , Organometallic Compounds/pharmacology , Salicylates/pharmacology , Animals , Gene Expression Regulation , HSP70 Heat-Shock Proteins/genetics , Liver/metabolism , Male , Oxidation-Reduction , RNA, Messenger/metabolism , Rats , Rats, Inbred F344 , Time FactorsABSTRACT
Exposure of mice to the herbicide paraquat has been demonstrated to result in the selective loss of dopaminergic neurons of the substantia nigra, pars compacta (SNpc) akin to what is observed in Parkinson disease (PD). In this study, we investigate the efficacy of two synthetic superoxide dismutase/catalase mimetics (EUK-134 and EUK-189) in protecting against paraquat-induced dopaminergic cell death in both the rat dopaminergic cell line 1RB3AN27 (N27) and primary mesencephalic cultures in vitro and in adult mice in vivo. Our data demonstrate that pretreatment with either EUK-134 or EUK-189 significantly attenuates paraquat-induced neurotoxicity in vitro in a concentration-dependent manner. Furthermore, systemic administration of EUK-189 decreases paraquat-mediated SNpc dopaminergic neuronal cell death in vivo. These findings support a role for oxidative stress in paraquat-induced neurotoxicity and suggest novel therapeutic approaches for neurodegenerative disorders associated with oxidative stress such as PD.
Subject(s)
Catalase/chemistry , Dopamine/metabolism , Neurons/drug effects , Neurons/pathology , Neuroprotective Agents/pharmacology , Paraquat/pharmacology , Parkinson Disease/pathology , Substantia Nigra/pathology , Superoxide Dismutase/chemistry , Animals , Antioxidants/pharmacology , Blotting, Western , Catalase/metabolism , Cells, Cultured , Immunohistochemistry , Male , Manganese/metabolism , Manganese Compounds/chemistry , Mice , Mice, Inbred C57BL , Microscopy, Fluorescence , Models, Chemical , Neurons/metabolism , Organometallic Compounds/pharmacology , Oxidative Stress , Rats , Salicylates/pharmacology , Superoxide Dismutase/metabolism , Time FactorsABSTRACT
Alzheimer disease is characterized by cerebral Abeta deposition, which we have recently discovered occurs also in the lens as cataracts in Alzheimer disease patients. Here we report the presence of significantly increased cataracts in the lenses of an Abeta-transgenic mouse model for Alzheimer disease and their amelioration upon treatment with EUK-189, a synthetic SOD/catalase mimetic. These data support an oxidative etiology for AD-associated lens cataracts and their potential to be treated preventatively with antioxidants.
Subject(s)
Alzheimer Disease/metabolism , Amyloid beta-Peptides/chemistry , Antioxidants/pharmacology , Catalase/pharmacology , Cataract/drug therapy , Cataract/metabolism , Organometallic Compounds/pharmacology , Salicylates/pharmacology , Animals , Free Radicals , Humans , Mice , Mice, Transgenic , Models, Chemical , Oxygen/chemistry , Superoxide Dismutase/metabolismABSTRACT
A reduction in stress tolerance is a hallmark of the aging process, and the lowered functional capacity observed in aged organisms is associated with an increased rate of oxidative stress and a greater susceptibility of aged tissues to oxidative injury. In this report, we show that chronic systemic administration of a superoxide dismutase (SOD)/catalase mimetic (EUK-189), delivered over a 1 month period via osmotic pump, prevents heat stress-induced liver injury by dramatically decreasing oxidative damage in aged animals. Widespread liver injury was present in old but not young vehicle-treated rats in response to a 2 day heating protocol. However, SOD/catalase mimetic treatment markedly decreased the hyperthermia-induced liver injury associated in old animals. The reversal of damage with EUK-189 was associated with an improvement in intracellular redox status and a striking reduction in hepatocellular lipid peroxidation. EUK-189 treatment also blocked the activation of activator protein-1 (AP-1), which is a redox-sensitive early response transcription factor involved in the regulation of cellular stress responses. These results demonstrate that oxidative stress plays a unique role in age-related hyperthermic injury and suggest that therapeutic strategies aimed at improving redox potential, such as chronic SOD/catalase mimetic treatment, can prevent the oxidative-mediated damage associated with environmental stress.