ABSTRACT
Caerulomycin A (CaeA), isolated from actinomycetes, has a featured 2,2'-bipyridine core structure. Based on the results of in silico drug-protein docking analysis, CaeA shows potential ligands for interacting with both tubulin and DNA topoisomerase I (Topo-1). The result was confirmed by cell-free tubulin polymerization assay and Topo-1 activity assay. In vitro assays also demonstrated that CaeA increases the polymerization of tubulin and increases cell size. In addition, CaeA inhibits cell viability and growth of various cancer cells, yet exhibits low cytotoxicity. CaeA also affects paclitaxel-resistant cancer cells and synergizes the effect with paclitaxel in reducing cancer cell colony formation rate. In vivo experiments confirm the effect of CaeA on reducing tumor size and weight in nude mouse inoculated with tumor cells with no noticeable side effects. Taken together, our data demonstrate that CaeA is a potential potent agent for cancer treatment through tubulin and Topo-1 dual-targeting with little side effects.
Subject(s)
Antineoplastic Agents , Neoplasms , 2,2'-Dipyridyl/pharmacology , 2,2'-Dipyridyl/therapeutic use , Animals , Antineoplastic Agents/therapeutic use , Cell Line, Tumor , Cell Proliferation , Drug Screening Assays, Antitumor , Mice , Neoplasms/drug therapy , Paclitaxel/pharmacology , Pyridines , TubulinABSTRACT
Transforming growth factor ß-induced protein (TGFBIp) is an extracellular matrix protein; its expression by several cell types is greatly increased by TGF-ß. TGFBIp is released by primary human umbilical vein endothelial cells (HUVECs) and functions as a mediator of experimental sepsis. 2,2'-Bipyridine-containing natural products are generally accepted to have antimicrobial, cytotoxic and anti-inflammatory properties. We hypothesized that a 2,2'-bipyridine containing natural product, collismycin C, could reduce TGFBIp-mediated severe inflammatory responses in human endothelial cells and mice. Here we investigated the effects and underlying mechanisms of collismycin C against TGFBIp-mediated septic responses. Collismycin C effectively inhibited lipopolysaccharide-induced release of TGFBIp and suppressed TGFBIp-mediated septic responses. In addition, collismycin C suppressed TGFBIp-induced sepsis lethality and pulmonary injury. This suppression of TGFBIp-mediated and CLP-induced septic responses indicates that collismycin C is a potential therapeutic agent for various severe vascular inflammatory diseases, with inhibition of the TGFBIp signaling pathway as the mechanism of action.
Subject(s)
2,2'-Dipyridyl/therapeutic use , Anti-Inflammatory Agents/therapeutic use , Extracellular Matrix Proteins/therapeutic use , Inflammation/drug therapy , Sepsis/drug therapy , Transforming Growth Factor beta/therapeutic use , 2,2'-Dipyridyl/pharmacology , Animals , Extracellular Matrix Proteins/pharmacology , Humans , Mice , Mice, Inbred C57BL , Transforming Growth Factor beta/pharmacologyABSTRACT
BACKGROUND: The chronic phase of Spinal Cord (SC) injury is characterized by the presence of a hostile microenvironment that causes low activity and a progressive decline in neurological function; this phase is non-compatible with regeneration. Several treatment strategies have been investigated in chronic SC injury with no satisfactory results. OBJECTIVE- In this proof-of-concept study, we designed a combination therapy (Comb Tx) consisting of surgical glial scar removal plus scar inhibition, accompanied with implantation of mesenchymal stem cells (MSC), and immunization with neural-derived peptides (INDP). METHODS: This study was divided into three subsets, all in which Sprague Dawley rats were subjected to a complete SC transection. Sixty days after injury, animals were randomly allocated into two groups for therapeutic intervention: control group and animals receiving the Comb-Tx. Sixty-three days after treatment we carried out experiments analyzing motor recovery, presence of somatosensory evoked potentials, neural regeneration-related genes, and histological evaluation of serotoninergic fibers. RESULTS: Comb-Tx induced a significant locomotor and electrophysiological recovery. An increase in the expression of regeneration-associated genes and the percentage of 5-HT+ fibers was noted at the caudal stump of the SC of animals receiving the Comb-Tx. There was a significant correlation of locomotor recovery with positive electrophysiological activity, expression of GAP43, and percentage of 5-HT+ fibers. CONCLUSION: Comb-Tx promotes motor and electrophysiological recovery in the chronic phase of SC injury subsequent to a complete transection. Likewise, it is capable of inducing the permissive microenvironment to promote axonal regeneration.
Subject(s)
Cicatrix/surgery , Combined Modality Therapy/methods , Mesenchymal Stem Cell Transplantation , Recovery of Function/drug effects , Recovery of Function/immunology , Spinal Cord Injuries , 2,2'-Dipyridyl/therapeutic use , Animals , Evoked Potentials/physiology , Female , Freund's Adjuvant/therapeutic use , Gene Expression/drug effects , Motor Activity/drug effects , Nerve Regeneration/drug effects , Rats , Spinal Cord Injuries/drug therapy , Spinal Cord Injuries/physiopathology , Spinal Cord Injuries/surgery , Spinal Cord Injuries/therapy , Tryptophan/analogs & derivatives , Tryptophan/therapeutic useABSTRACT
Cancer is the second leading cause of death worldwide and there is epidemiological evidence that demonstrates this tendency is emerging. Naringenin (NGEN) is a trihydroxyflavanone that shows various biological effects such as antioxidant, anticancer, anti-inflammatory, and antiviral activities. It belongs to flavanone class, which represents flavonoids with a C6-C3-C6 skeleton. Flavonoids do not exhibit sufficient activity to be used for chemotherapy, however they can be chemically modified by complexation with metals such as copper (Cu) (II) for instance, in order to be applied for adjuvant therapy. This study investigated the effects of Cu(II) and 2,2'-bipyridine complexation with naringenin on MDA-MB-231 cells. We demonstrated that naringenin complexed with Cu(II) and 2,2'-bipyridine (NGENCuB) was more efficient inhibiting colony formation, proliferation and migration of MDA-MB-231 tumor cells, than naringenin (NGEN) itself. Furthermore, we verified that NGENCuB was more effective than NGEN inhibiting pro-MMP9 activity by zymography assays. Finally, through flow cytometry, we showed that NGENCuB is more efficient than NGEN inducing apoptosis in MDA-MB-231 cells. These results were confirmed by gene expression analysis in real time PCR. We observed that NGENCuB upregulated the expression of pro-apoptotic gene caspase-9, but did not change the expression of caspase-8 or anti-apoptotic gene Bcl-2. There are only few works investigating the effects of Cu(II) complexation with naringenin on tumor cells. To the best of our knowledge, this is the first work describing the effects of Cu(II) complexation of a flavonoid on MDA-MB-231 breast tumor cells.
Subject(s)
2,2'-Dipyridyl/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Breast Neoplasms/prevention & control , Coordination Complexes/pharmacology , Copper/pharmacology , Flavanones/pharmacology , 2,2'-Dipyridyl/chemistry , 2,2'-Dipyridyl/therapeutic use , Antineoplastic Agents, Phytogenic/therapeutic use , Breast Neoplasms/pathology , Cell Line, Tumor , Cell Proliferation/drug effects , Chemoprevention , Coordination Complexes/therapeutic use , Copper/chemistry , Copper/therapeutic use , Drug Evaluation, Preclinical , Drug Synergism , Female , Flavanones/therapeutic use , HumansSubject(s)
2,2'-Dipyridyl/analogs & derivatives , Immunosuppressive Agents/therapeutic use , 2,2'-Dipyridyl/chemistry , 2,2'-Dipyridyl/therapeutic use , Actinobacteria/chemistry , Animals , Cell Proliferation , Immune System , Immunosuppressive Agents/chemistry , Inflammation , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , T-Lymphocytes/immunology , Tacrolimus/therapeutic useABSTRACT
We investigated the efficacy of the ferrous iron (Fe(2+)) chelator 2,2'-dipyridyl (DP) to attenuate cerebral vasospasm after subarachnoid hemorrhage (SAH). Thirty-six New Zealand white rabbits were randomly assigned to four groups: untreated control, SAH, SAH + dimethyl sulfoxide (DMSO) vehicle, and SAH + DP. SAH was induced by injection of autologous blood into the cisterna magna and then DP or vehicle was infused into the cistern magna for 5 days (20 mg/kg/day or an equal volume of DMSO). Neurological deficit score (NDS) was used to assess neurological function and cerebral angiography to measure basilar artery (BA) diameter following SAH. TUNEL staining was used to detect BA endothelial cell apoptosis, and immunohistochemistry and Western blotting to assess changes in caspase-3 protein levels 5 days post-SAH. The SAH + DP group had a significantly larger mean BA diameter and lower mean NDS post-SAH compared to the SAH + DMSO and SAH groups (p < 0.05). TUNEL-positive cell numbers and caspase-3 levels were significantly reduced in BA endothelial cells of the SAH + DP group as compared to the SAH and SAH + DMSO groups (p < 0.05). The iron chelator DP reduced vasospasm and neurological sequelae in rabbits, likely by chelating the Fe(2+) in oxyhemoglobin and reducing oxidative stress-induced endothelial cell apoptosis.
Subject(s)
2,2'-Dipyridyl/therapeutic use , Iron Chelating Agents/therapeutic use , Nervous System Diseases/drug therapy , Subarachnoid Hemorrhage/complications , Vasospasm, Intracranial/drug therapy , Vasospasm, Intracranial/etiology , Angiography, Digital Subtraction , Animals , Apoptosis/drug effects , Basilar Artery/drug effects , Basilar Artery/pathology , Blood Transfusion, Autologous/adverse effects , Caspase 3/metabolism , Cerebral Angiography , Disease Models, Animal , Endothelial Cells/drug effects , Female , In Situ Nick-End Labeling , Male , Nervous System Diseases/etiology , Neurologic Examination , RabbitsABSTRACT
We investigated the ability of compounds interfering with iron metabolism to inhibit the growth of Acinetobacter baumannii. Iron restriction with transferrin or 2,2-bipyridyl significantly inhibited A. baumannii growth in vitro. Gallium nitrate alone was moderately effective at reducing A. baumannii growth but became bacteriostatic in the presence of serum or transferrin. More importantly, gallium nitrate treatment reduced lung bacterial burdens in mice. The use of gallium-based therapies shows promise for the control of multidrug-resistant A. baumannii.
Subject(s)
2,2'-Dipyridyl/pharmacology , Acinetobacter baumannii/drug effects , Acinetobacter baumannii/pathogenicity , Gallium/pharmacology , Transferrin/pharmacology , 2,2'-Dipyridyl/therapeutic use , Acinetobacter Infections/drug therapy , Animals , Gallium/therapeutic use , Iron Chelating Agents/pharmacology , Iron Chelating Agents/therapeutic use , Mice , Transferrin/therapeutic useABSTRACT
Previous studies have indicated that 2,2'-dipyridyl, a lipid-soluble ferrous iron chelator, can reduce brain injury after cerebral ischemia and reduce cerebral vasospasm after subarachnoid hemorrhage. In this study, we examined the efficacy of 2,2'-dipyridyl after intracerebral hemorrhage (ICH) in 12-month-old mice. ICH was modeled by intrastriatal injection of collagenase or autologous whole blood. 2,2'-Dipyridyl or vehicle was administered intraperitoneally 2h before ICH (pretreatment) or 6h after ICH (post-treatment) and then once daily for up to 3 days. Mice in the pretreatment group were sacrificed 1 or 3 days after ICH and examined for iron deposition, neuronal death, oxidative stress, microglial/astrocyte activation, neutrophil infiltration, and white matter damage. Mice in the post-treatment group were examined for brain lesion volume and edema on day 3 and for neurologic deficits on days 1, 3, and 28 after ICH. Pretreatment with 2,2'-dipyridyl decreased iron accumulation and neuronal death, attenuated production of reactive oxygen species, reduced microglial activation without affecting astrocytes or neutrophil infiltration, and attenuated white matter damage. Post-treatment reduced brain lesion volume and edema and improved neurologic function. These results indicate that the lipid-soluble ferrous iron chelator 2,2'-dipyridyl can reduce brain injury and improve functional outcome after ICH.
Subject(s)
2,2'-Dipyridyl/therapeutic use , Chelating Agents/therapeutic use , Intracranial Hemorrhages/drug therapy , Iron/metabolism , Neurons/drug effects , 2,2'-Dipyridyl/pharmacology , Animals , Cell Death/drug effects , Chelating Agents/pharmacology , Intracranial Hemorrhages/metabolism , Intracranial Hemorrhages/pathology , Mice , Neuroglia/drug effects , Neuroglia/metabolism , Neuroglia/pathology , Neurons/metabolism , Neurons/pathology , Treatment OutcomeABSTRACT
Whereas iron chelators were shown to induce neuroprotection against brain injury, the effect of iron chelators on ischemia-induced damage of cerebral endothelium is largely unknown. Our objective was to explore the endothelioprotective effect of the lipophilic iron chelator dipyridyl (DP) (i) in vitro on the death of cerebral endothelial cells (CECs) subjected to intracellular iron loading and (ii) in vivo on the ischemia-induced blood-brain barrier (BBB) disruption. When given shortly after iron exposure or brain ischemia, DP prevented the death of CECs and diminished BBB disruption, respectively, whereas a delayed administration of DP was associated with a lower CECs protection. Interestingly, when given preventively, DP also abrogated the death of CECs and reduced BBB disruption. However, a long delay between DP treatment and iron exposure led to a higher protection, suggesting a preconditioning effect of DP. Accordingly, prevention of hydroxyl radical formation through iron chelation cannot explain alone the beneficial effect of preventive DP treatment. Our findings showing that DP failed to induce the potentially cytoprotective proteins, heme oxygenase-1 and manganese superoxide dismutase, suggest that other proteins participate to the preconditioning effect of DP. To conclude, the curative and preventive effects of DP evidenced in this study suggest that iron chelation therapy represents a favorable and effective approach to increase BBB resistance towards ischemic injury.
Subject(s)
2,2'-Dipyridyl/therapeutic use , Blood-Brain Barrier/drug effects , Brain Ischemia/pathology , Brain Ischemia/prevention & control , Endothelial Cells/drug effects , Iron Chelating Agents/therapeutic use , Animals , Blood-Brain Barrier/physiopathology , Cell Death/drug effects , Cells, Cultured , Disease Models, Animal , Dose-Response Relationship, Drug , Gene Expression Regulation/drug effects , Heme Oxygenase-1/metabolism , Iron/metabolism , Male , Mice , Statistics, Nonparametric , Superoxide Dismutase/metabolism , Time Factors , TransfectionABSTRACT
Traumatic injury of the central nervous system results in formation of a collagenous basement membrane-rich fibrous scar in the lesion centre. Due to accumulation of numerous axon-growth inhibitory molecules the lesion scar is considered a major impediment for axon regeneration. Following transection of the dorsal corticospinal tract (CST) at thoracic level 8 in adult rats, transient suppression of collagenous scarring in the lesion zone by local application of a potent iron chelator and cyclic adenosine monophosphate resulted in the delay of fibrous scarring. Treated animals displayed long-distance growth of CST axons through the lesion area extending for up to 1.5-2 cm into the distal cord. In addition, the treatment showed a strong neuroprotective effect, rescuing cortical motoneurons projecting into the CST that normally die (30%) after thoracic axotomy. Further, anterogradely traced CST axons regenerated through both grey and white matter and developed terminal arborizations in grey matter regions. In contrast to controls, injured animals receiving treatment showed significant functional recovery in the open field, in the horizontal ladder and in CatWalk locomotor tasks. We conclude that the fibrous lesion scar plays a pivotal role as a growth barrier for regenerating axons in adult spinal cord and that a delay in fibrotic scarring by local inhibition of collagen biosynthesis and basement membrane deposition is a promising and unique therapeutic strategy for treating human spinal trauma.
Subject(s)
Cicatrix/prevention & control , Nerve Regeneration/physiology , Pyramidal Tracts/pathology , Recovery of Function , Somatosensory Cortex/physiopathology , Spinal Cord Injuries/pathology , 2',3'-Cyclic-Nucleotide Phosphodiesterases/metabolism , 2,2'-Dipyridyl/analogs & derivatives , 2,2'-Dipyridyl/therapeutic use , 8-Bromo Cyclic Adenosine Monophosphate/therapeutic use , Animals , Antigens/metabolism , Axons/pathology , Axons/physiology , Behavior, Animal , Biotin/analogs & derivatives , Biotin/metabolism , Cell Count/methods , Cicatrix/etiology , Collagen Type IV/metabolism , Dextrans/metabolism , Female , Ferrous Compounds/therapeutic use , Functional Laterality , Glial Fibrillary Acidic Protein/metabolism , Immunohistochemistry/methods , Motor Activity/drug effects , Motor Activity/physiology , Proteoglycans/metabolism , Rats , Rats, Wistar , Spinal Cord Injuries/therapy , Stilbamidines , Time FactorsABSTRACT
Two different forms of cell death have been distinguished morphologically following cerebral ischaemia: necrotic and apoptotic cell death. The aim of this study was to investigate the contribution of apoptosis to ischaemic damage by carefully depicting the temporal and spatial neuronal death following focal ischaemia. For this purpose, rats were subjected to chemical photothrombosis, and histological and biochemical analyses were performed over a period of 24 h after the onset of ischaemia. In addition, the effects of the lipophilic antioxidant iron chelator 2,2'-dipyridyl (DP) were evaluated 24 h after photothrombosis when the lesion volume was maximal. Our results showed two separate waves of neuronal death. In the first wave, shrunken dark neurons were massively present as early as 2 h after photothrombosis in the infarct core. From this initial neuronal abnormal population, progressive and time-dependent changes of both necrotic and apoptotic cell death were observed, leading to ghost neurons and apoptotic bodies after 24 h. The extension of the lesion coincided with a second wave of cell death. Massive and rapid neuronal loss occurred at the infarct border, which appeared as a sharply demarcated pale region. Procaspase and poly(ADP-ribose) polymerase-1 (PARP-1) cleavages were also detected in the infarct core and surrounding damaged tissue. DP treatment markedly blocked the enlargement of the lesion, the infarct border being rescued from infarction. Furthermore, a large decrease of apoptotic bodies was associated with a significant drop of caspase and PARP-1 cleavages, suggesting that the protective effect of DP closely correlates with limitation of apoptosis expansion.
Subject(s)
2,2'-Dipyridyl/therapeutic use , Apoptosis/physiology , Brain Ischemia/pathology , Chelating Agents/therapeutic use , Animals , Blotting, Western/methods , Brain Chemistry/physiology , Brain Chemistry/radiation effects , Brain Infarction/pathology , Brain Infarction/physiopathology , Brain Infarction/prevention & control , Brain Ischemia/physiopathology , Caspase 3 , Caspase 9 , Caspases/metabolism , DNA Fragmentation/drug effects , DNA Fragmentation/physiology , Disease Models, Animal , Fluorescent Antibody Technique/methods , Functional Laterality/drug effects , Functional Laterality/physiology , Intracranial Thrombosis/pathology , Intracranial Thrombosis/physiopathology , Male , Poly (ADP-Ribose) Polymerase-1 , Poly(ADP-ribose) Polymerases/metabolism , Rats , Rats, Wistar , Time FactorsABSTRACT
We examined the efficacy of the liposoluble iron chelator 2,2'-dipyridyl (DP) in reducing histological damage in rats submitted to cerebral ischemia and the mechanisms involved in the potential cytoprotection. For this purpose, DP (20 mg/kg, i.p.) was administered 15 min before and 1 h after induction of cortical photothrombotic vascular occlusion in rat. Histological studies were performed to assess infarct volume (at days 1 and 3 postischemia) and astromicroglial activation (at day 3 postischemia). Damage to endothelial and neuronal cells was evaluated at day 1 postischemia by quantitative measurements of Evans Blue extravasation and N-acetylaspartate levels, respectively. Cerebral blood flow was recorded in the ischemic core by laser-Doppler flowmetry within the 15 min to 2 h period after photothrombosis. At 4-h postischemia, radical oxygen species (ROS) production was evaluated by measuring brain glutathione concentrations. The cortical expression of the proteins heme oxygenase-1 (HO-1) and hypoxia-inducible factor-1alpha (HIF-1alpha) was analyzed by Western blotting at day 1 postischemia. Infarct volume and ischemic damage to endothelial and neuronal cells were significantly reduced by DP treatment. This cytoprotection was associated with a reduction in ROS production, perfusion deficits, and astrocytic activation. DP treatment also resulted in significant changes in HO-1 (+100%) and HIF-1alpha (-50%) protein expression at the level of the ischemic core. These results report the efficacy of the liposoluble iron chelator DP in reducing histological damage induced by permanent focal ischemia.
Subject(s)
2,2'-Dipyridyl/therapeutic use , Brain Ischemia/drug therapy , Iron Chelating Agents/therapeutic use , Stroke/drug therapy , 2,2'-Dipyridyl/chemistry , Animals , Astrocytes/drug effects , Astrocytes/pathology , Brain Ischemia/pathology , Cell Survival/drug effects , Cerebral Cortex/pathology , Cerebral Infarction/pathology , Cerebrovascular Circulation/drug effects , DNA-Binding Proteins/metabolism , Glutathione/metabolism , Heme Oxygenase (Decyclizing)/biosynthesis , Heme Oxygenase-1 , Hypoxia-Inducible Factor 1 , Hypoxia-Inducible Factor 1, alpha Subunit , Iron Chelating Agents/chemistry , Lasers , Male , Neuroglia/drug effects , Neuroglia/pathology , Neurons/pathology , Nuclear Proteins/metabolism , Rats , Rats, Wistar , Reactive Oxygen Species/metabolism , Stroke/pathology , Thrombosis/drug therapy , Thrombosis/pathology , Transcription Factors/metabolismABSTRACT
Identification of a vanadium compound with the highest efficacy and least toxicity is the main scientific problem in diabetes treatment. All vanadium complexes, both inorganic and organic, apart from improving physiological and biochemical diabetic parameters, show more or less toxic effects in living organisms. For this reason we decided to test a new vanadium compound: bis(2,2'-bipyridine)oxovanadium(IV), [VO(bpy)2], not used or described so far. This paper stressed morphological alterations of rat liver Golgi apparatus originated from control or streptozotocin (STZ)-diabetic rats treated with 1.8 mmol VO(bpy)2 solution in 0.5% NaCl as drinking liquid for 7 days and compared them with a parallel biochemical study. There was a correlation between the activity of Golgi marker enzyme i.e. galactosyl transferase and morphology of this organelle. In control rats treatment with VO(bpy)2 caused drastic changes, in many cases leading to a complete destruction of liver Golgi apparatus. In STZ-diabetic liver of rats treated with VO(bpy)2 the Golgi apparatus showed characteristic of untreated diabetes arching or even twisting of stack cisternae but improvement of the secretory activity (dilatation of cisternae edges, some secretory vacuoles and vesicles). In our opinion, the parallel action of two drugs: STZ combined with VO(bpy)2, relieves or even eliminate harmful effects of each compound alone.
Subject(s)
2,2'-Dipyridyl/pharmacology , Diabetes Mellitus, Experimental/pathology , Golgi Apparatus/ultrastructure , Hepatocytes/drug effects , Organometallic Compounds/pharmacology , Vanadium , 2,2'-Dipyridyl/therapeutic use , Animals , Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Experimental/enzymology , Female , Golgi Apparatus/drug effects , Golgi Apparatus/enzymology , Golgi Apparatus/metabolism , Hepatocytes/ultrastructure , Liver/drug effects , Liver/metabolism , Liver/pathology , Microscopy, Electron , Organometallic Compounds/therapeutic use , Rats , Rats, Wistar , Streptozocin , Transferases/metabolismABSTRACT
Following traumatic injury, the formation of a glial scar and deposition of extracellular matrix (ECM) contributes to the regeneration failure in the adult mammalian central nervous system (CNS). Using a postcommissural fornix transection as a brain lesion model in rat, we have previously shown that the collagenous basement membrane (BM) at the lesion site is a major impediment for axon regeneration. Deposition of BM in this lesion model can be delayed by administration of the iron chelator 2,2'-bipyridine (BPY), an inhibitor of prolyl 4-hydroxylase (PH), a key enzyme of collagen biosynthesis. To examine whether this potential therapeutic approach is transferable to other CNS regions, we have chosen the mechanically lesioned rat spinal cord to investigate the effects of BPY administration on BM formation. Due to the close proximity of the lesion zone to meningeal fibroblasts, a cell-type secreting large amounts of collagen IV, BM deposition was much more extensive in the spinal cord than in the brain lesion. Neither immediate injections nor continuous application of BPY resulted in a detectable reduction of BM formation in the spinal cord. Only a combination of anti-scarring treatments including (i) injection of the more potent PH inhibitor [2,2'-bipyridine]-5,5'-dicarboxylic acid (BPY-DCA), (ii) selective inhibition of fibroblast proliferation and ECM production by 8-Br-cAMP, and (iii) continuous application of BPY-DCA, reduced the lesion-induced BM significantly. The present results clearly demonstrate, that the exclusive application of BPY according to a protocol designed for treatment of brain lesions is not sufficient to reduce BM formation in the lesioned adult rat spinal cord.
Subject(s)
2,2'-Dipyridyl/pharmacology , Basement Membrane/drug effects , Cicatrix/prevention & control , Collagen Type IV/antagonists & inhibitors , Indicators and Reagents/pharmacology , Nerve Regeneration/drug effects , Spinal Cord Injuries/drug therapy , Spinal Cord/drug effects , 2,2'-Dipyridyl/therapeutic use , 8-Bromo Cyclic Adenosine Monophosphate/pharmacology , 8-Bromo Cyclic Adenosine Monophosphate/therapeutic use , Animals , Basement Membrane/metabolism , Basement Membrane/pathology , Chelating Agents/pharmacology , Chelating Agents/therapeutic use , Cicatrix/etiology , Cicatrix/physiopathology , Collagen Type IV/biosynthesis , Disease Models, Animal , Dose-Response Relationship, Drug , Drug Administration Schedule , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/therapeutic use , Extracellular Matrix/drug effects , Extracellular Matrix/metabolism , Female , Gliosis/etiology , Gliosis/physiopathology , Gliosis/prevention & control , Iron/metabolism , Male , Nerve Regeneration/physiology , Procollagen-Proline Dioxygenase/antagonists & inhibitors , Procollagen-Proline Dioxygenase/metabolism , Rats , Rats, Wistar , Spinal Cord/physiopathology , Spinal Cord/surgery , Spinal Cord Injuries/complications , Spinal Cord Injuries/physiopathology , Treatment OutcomeABSTRACT
To evaluate the contribution of cellular dysfunction and neuronal loss to brain N-acetylaspartate (NAA) depletion, NAA was measured in brain tissue by HPLC and UV detection in rats subjected to cerebral injury, associated or not with cell death. When lesion was induced by intracarotid injection of microspheres, the fall in NAA was related to the degree of embolization and to the severity of brain oedema. When striatal lesion was induced by local injection of malonate, the larger the lesion volume, the higher the NAA depletion. However, reduction of brain oedema and striatal lesion by treatment with the lipophilic iron chelator dipyridyl (20 mg/kg, 1 h before and every 8 h after embolization) and the inducible nitric oxide synthase inhibitor aminoguanidine (100 mg/kg given 1 h before malonate and then every 9 h), respectively, failed to ameliorate the fall in NAA. Moreover, after systemic administration of 3-nitropropionic acid, a marked reversible fall in NAA striatal content was observed despite the lack of tissue necrosis. Overall results show that cellular dysfunction can cause higher reductions in NAA level than neuronal loss, thus making of NAA quantification a potential tool for visualizing the penumbra area in stroke patients.
Subject(s)
Aspartic Acid/analogs & derivatives , Aspartic Acid/analysis , Brain Chemistry , Brain Edema/metabolism , Brain Ischemia/metabolism , 2,2'-Dipyridyl/administration & dosage , 2,2'-Dipyridyl/pharmacology , 2,2'-Dipyridyl/therapeutic use , Acute Disease , Animals , Biomarkers , Brain Edema/etiology , Brain Edema/pathology , Brain Ischemia/etiology , Brain Ischemia/pathology , Carotid Arteries , Cell Death , Chromatography, High Pressure Liquid , Corpus Striatum/drug effects , Corpus Striatum/metabolism , Corpus Striatum/pathology , Disease Models, Animal , Enzyme Inhibitors/pharmacology , Guanidines/administration & dosage , Guanidines/pharmacology , Guanidines/therapeutic use , Injections, Intra-Arterial , Iron Chelating Agents/administration & dosage , Iron Chelating Agents/pharmacology , Iron Chelating Agents/therapeutic use , Male , Malonates/toxicity , Microspheres , Mitochondria/drug effects , Nerve Tissue Proteins/antagonists & inhibitors , Neuroprotective Agents/administration & dosage , Neuroprotective Agents/pharmacology , Neuroprotective Agents/therapeutic use , Nitric Oxide/metabolism , Nitric Oxide Synthase/antagonists & inhibitors , Nitro Compounds , Oxidative Stress , Premedication , Propionates/toxicity , Rats , Rats, Wistar , Spectrophotometry, Ultraviolet , Succinate Dehydrogenase/antagonists & inhibitorsABSTRACT
The production of specific extracellular matrix molecules is upregulated following injury to the adult CNS, and some of these molecules have been postulated to inhibit axonal regeneration. In particular, the deposition of collagen in conjunction with basal lamina formation has been correlated with the failure of CNS axons to extend beyond sites of injury. In the present experiment, the spatial and temporal distribution of fibrillar collagen type III and the main constituents of basal lamina (collagen type IV and laminin) were characterized after defined lesions of the adult spinal cord at cervical and thoracic levels. The deposition of collagen was then blocked in animals undergoing defined mid-thoracic spinal cord lesions by administration of the iron chelator 2,2'-bipyridine, and subsequent effects on corticospinal axonal growth were examined. At time points from 1 to 6 weeks postinjury, collagen and laminin were deposited at spinal cord lesion sites as a dense matrix at the host-lesion interface that extended for short distances into the surrounding spinal cord parenchyma. The failure of corticospinal axons to grow beyond the lesioned region correlated spatially and temporally with collagen III formation and basal lamina production. However, successful blockade of collagen and basal lamina formation with 2,2'-bipyridine injections failed to enhance corticospinal axon regeneration or sprouting. These results suggest either that collagen and basal lamina formation after CNS injury do not contribute to corticospinal axonal growth failure or, more likely, that molecules in addition to collagen and basal lamina contribute to axonal growth failure and must be collectively blocked to promote corticospinal regeneration.
Subject(s)
2,2'-Dipyridyl/therapeutic use , Axons/pathology , Basement Membrane/pathology , Cicatrix/prevention & control , Collagen/physiology , Extracellular Matrix/pathology , Iron Chelating Agents/therapeutic use , Nerve Regeneration , Pyramidal Tracts/physiology , Spinal Cord Injuries/pathology , 2,2'-Dipyridyl/administration & dosage , 2,2'-Dipyridyl/pharmacology , Animals , Basement Membrane/drug effects , Collagen/analysis , Collagen/biosynthesis , Extracellular Matrix/drug effects , Injections, Intralesional , Iron Chelating Agents/administration & dosage , Iron Chelating Agents/pharmacology , Laminin/biosynthesis , Meninges/pathology , Rats , Rats, Inbred F344 , Thoracic VertebraeABSTRACT
OBJECT: Oxyhemoglobin (HbO2) causes vasospasm after subarachnoid hemorrhage (SAH). The most likely spasmogenic component of HbO2 is iron. Various iron chelators, such as deferoxamine, have prevented vasospasm in vivo with limited success. However, only chelators of iron in the ferric state have been studied in animal models of vasospasm after SAH. Because free radical formation requires the ferrous (Fe++) moiety and Fe++ is a potent binder of the vasodilator nitric oxide, the authors hypothesized that iron in the ferrous state causes vasospasm and that chelators of Fe++, such as 2,2'-dipyridyl, may prevent vasospasm. This study was undertaken to investigate the influence of 2,2'-dipyridyl on vasospasm after induction of SAH in a primate model. METHODS: Twelve cynomolgus monkeys were randomly divided into two groups and then both groups underwent placement of an arterial autologous blood clot in the subarachnoid space around the right middle cerebral artery (MCA). The five animals in the control group received intravenously administered saline and the seven treated animals received intravenously administered chelator (2,2'-dipyridyl) for 14 days. Sequential arteriography for assessment of MCA diameter was performed before and on the 7th day after SAH. CONCLUSIONS: Prevention of cerebral vasospasm by means of treatment with continuous intravenous administration of 2,2'-dipyridyl is reported in a primate model of SAH. This result provides insight into the possible mechanism of delayed vasospasm after aneurysmal SAH and provides a potential preventive therapy for it.
Subject(s)
2,2'-Dipyridyl/therapeutic use , Ferrous Compounds/antagonists & inhibitors , Iron Chelating Agents/therapeutic use , Ischemic Attack, Transient/prevention & control , Subarachnoid Hemorrhage/drug therapy , 2,2'-Dipyridyl/adverse effects , Animals , Cerebral Angiography , Female , Injections, Intravenous , Iron Chelating Agents/adverse effects , Ischemic Attack, Transient/diagnostic imaging , Liver/drug effects , Liver/pathology , Macaca fascicularis , Male , Time Factors , Transferrin/analysisABSTRACT
1. 2,2'-Bipyridyl-6-carbothioamide (BPYTA) is a synthesized compound with chelating properties demonstrating in vitro and in vivo antitumor activity. 2. The BPYTA cytotoxic effect is mainly due to the inhibition of ribonucleotide reductase (RR), a key enzyme in proliferating cells. The active form of BPYTA is its iron chelate [BPYTA-Fe(II), molar ratio 2:1], which destroys the tyrosyl radical of RR small subunit (R2). 3. The copper chelate of BPYTA [BPYTA-Cu(II), molar ratio 2:1] also has antiproliferative activity, but RR is not the only intracellular target. 4. BPYTA potently synergizes in vitro with hydroxyurea, the most widely used R2 inhibitor.
Subject(s)
2,2'-Dipyridyl/analogs & derivatives , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Chelating Agents/pharmacology , Chelating Agents/therapeutic use , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/therapeutic use , Ribonucleotide Reductases/antagonists & inhibitors , 2,2'-Dipyridyl/chemistry , 2,2'-Dipyridyl/pharmacology , 2,2'-Dipyridyl/therapeutic use , Animals , Antineoplastic Agents/chemistry , Enzyme Inhibitors/chemistry , Humans , Neoplasms/drug therapySubject(s)
2,2'-Dipyridyl/therapeutic use , Iron Chelating Agents/therapeutic use , Subarachnoid Hemorrhage/drug therapy , Animals , Cerebral Arteries , Humans , Ischemic Attack, Transient/physiopathology , Ischemic Attack, Transient/prevention & control , Primates , Subarachnoid Hemorrhage/physiopathologyABSTRACT
Ternary Cu(II) complexes with bidentate malonato- and heterocyclic amine ligands were tested with regard to cytotoxicity and potentiation of x-ray induced cell killing in V79 cells. Two lead complexes were also tested in a tumor assay using the MTG-B murine adenocarcinoma model growing in the flanks of female C3H/HeJ mice. One complex, [2,2'-bipyridyl malonatoCu(II)] (RL-5077), produced sensitizer enhancement ratios (SER's) of 1.8 (hypoxic conditions) and 1.0 (oxic conditions) in vitro when irradiation followed 1 hr exposure to the drug at 100 microM. When RL-5077 was administered at doses of 1/2 (11.65 mg/kg) or 1/4 (5.25 mg/kg) the maximum tolerated dose (MTD), 15 min prior to a locally delivered dose of 20 Gy, enhancement ratios (ER's) of 1.6 and 2, respectively, resulted. The second lead complex, [1,10 phenanthroline (malonato)Cu(II)hydrate] (RL-5027), produced SER's of 1.8 and 1.2 under hypoxic and oxic conditions, respectively, at a concentration of 25 microM. Injection of RL-5027 (5 mg/kg) resulted in toxicity without enhancement in combination with radiation. Analogues of these two complexes have been synthesized in an effort to optimize the potentiation of radiation effects while minimizing toxicity to drug alone and increasing water solubility of the drug. Further studies of the structure-activity relationship of Cu(II) ternary complexes using in vitro radiosensitization as the endpoint have identified four classes of ligands with varying biological activity and have supplied information about the effects of group substitution on solubility, toxicity, and radiation potentiation. This group of complexes represents a new class of radiopotentiators that deserves further investigation into its potential for clinical use.