RESUMEN
Activation of microglia and astrocytes following germinal matrix hemorrhage and intraventricular hemorrhage (GMH-IVH) plays a detrimental role in posthemorrhagic hydrocephalus (PHH). It is still unclear whether or how an interaction occurs between microglia and astrocytes in PHH. Here, we investigated the role of the C3/C3aR pathway in microglia and astrocyte interactions and whether C3/C3aR-targeted inhibition could alleviate PHH following GMH-IVH. A total of 152 Sprague-Dawley rats at postnatal day seven (P7) were enrolled in the study, and collagenase VII was used to induce GMH-IVH. Minocycline (45 mg/kg) was administered to inhibit microglial activation. Complement C3a peptide and C3aR antagonist (SB 290157, 10 mg/kg) were used to regulate the C3/C3aR pathway. As a result, the data demonstrated that periventricular C3aR+/Iba-1+ microglia and C3+/GFAP+ astrocytes were significantly increased in GMH-IVH pups at 28 days after surgery. Intranasal C3a peptide upregulated C3aR expression in microglia. Inhibition of microglia by minocycline decreased both C3+/GFAP+ astrocytes and the colocalization volume of Iba-1 and GFAP. In addition, intraperitoneally injected C3aRA alleviated the periventricular colocalization volume of microglia and astrocytes. Compared with vehicle-treated pups, the protein level of IL-1ß, IL-6 and TNF-α in cerebral spinal fluid and brain tissue at 28 days following GMH-IVH were reduced in C3aRA-treated pups. Moreover, hydrocephalus was alleviated, and long-term cognitive ability were improved in the C3aRA-treated group. Our data presented simultaneous periventricular astrogliosis and microgliosis of pups following GMH-IVH and proved their potential interaction through the C3/C3aR pathway, indicating C3aRA as a potential pharmacological treatment of PHH in neonates.
Asunto(s)
Arginina/análogos & derivados , Astrocitos/efectos de los fármacos , Compuestos de Bencidrilo/farmacología , Hemorragia Cerebral/tratamiento farmacológico , Hemorragia Cerebral/metabolismo , Complemento C3a/farmacología , Hidrocefalia/tratamiento farmacológico , Microglía/efectos de los fármacos , Receptores Acoplados a Proteínas G/efectos de los fármacos , Animales , Animales Recién Nacidos , Arginina/administración & dosificación , Arginina/farmacología , Compuestos de Bencidrilo/administración & dosificación , Hemorragia Cerebral/complicaciones , Hemorragia Cerebral Intraventricular/complicaciones , Hemorragia Cerebral Intraventricular/tratamiento farmacológico , Hemorragia Cerebral Intraventricular/metabolismo , Complemento C3a/administración & dosificación , Modelos Animales de Enfermedad , Hidrocefalia/etiología , Hidrocefalia/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores Acoplados a Proteínas G/antagonistas & inhibidoresRESUMEN
Intraventricular hemorrhage (IVH) results in periventricular inflammation, hypomyelination of the white matter, and hydrocephalus in premature infants. No effective therapy exists to prevent these disorders. Peroxisome proliferator activated receptor-γ (PPAR-γ) agonists reduce inflammation, alleviate free radical generation, and enhance microglial phagocytosis, promoting clearance of debris and red blood cells. We hypothesized that activation of PPAR-γ would enhance myelination, reduce hydrocephalus, and promote neurological recovery in newborns with IVH. These hypotheses were tested in a preterm rabbit model of IVH; autopsy brain samples from premature infants with and without IVH were analyzed. We found that IVH augmented PPAR-γ expression in microglia of both preterm human infants and rabbit kits. The treatment with PPAR-γ agonist or PPAR-γ overexpression by adenovirus delivery further elevated PPAR-γ levels in microglia, reduced proinflammatory cytokines, increased microglial phagocytosis, and improved oligodendrocyte progenitor cell (OPC) maturation in kits with IVH. Transcriptomic analyses of OPCs identified previously unrecognized PPAR-γ-induced genes for purinergic signaling, cyclic adenosine monophosphate generation, and antioxidant production, which would reprogram these progenitors toward promoting myelination. RNA-sequencing analyses of microglia revealed PPAR-γ-triggered down-regulation of several proinflammatory genes and transcripts having roles in Parkinson's disease and amyotrophic lateral sclerosis, contributing to neurological recovery in kits with IVH. Accordingly, PPAR-γ activation enhanced myelination and neurological function in kits with IVH. This also enhanced microglial phagocytosis of red blood cells but did not reduce hydrocephalus. Treatment with PPAR-γ agonist might enhance myelination and neurological recovery in premature infants with IVH.
Asunto(s)
Hemorragia Cerebral Intraventricular/metabolismo , Proteínas de la Mielina/biosíntesis , PPAR gamma/metabolismo , Sistemas de Transporte de Aminoácidos Acídicos/deficiencia , Sistemas de Transporte de Aminoácidos Acídicos/metabolismo , Animales , Animales Recién Nacidos , Antiportadores/deficiencia , Antiportadores/metabolismo , Hemorragia Cerebral Intraventricular/patología , Modelos Animales de Enfermedad , Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias/metabolismo , Humanos , Recien Nacido Prematuro , Microglía/metabolismo , Enfermedades Mitocondriales/metabolismo , Oligodendroglía/patología , PPAR gamma/agonistas , Trastornos Psicomotores/metabolismo , Conejos , Rosiglitazona/farmacología , Análisis de Secuencia de ARN/métodosRESUMEN
Background CD47, a glycoprotein on red blood cell membranes, inhibits phagocytosis via interaction with signal regulatory protein α on phagocytes. Our previous research has demonstrated that blocking CD47 accelerates hematoma clearance and reduces brain injury after intracerebral hemorrhage. The current study investigated whether phagocytosis or erythrocyte CD47 impacts hematoma resolution and hydrocephalus development after intraventricular hemorrhage (IVH). Methods Adult (3-month-old) male Fischer 344 rats were intraventricularly injected with 200 µl autologous blood, mixed with either CD47 blocking antibody or isotype IgG, or 200 µl saline as control. In subgroups of CD47 blocking antibody treated rats, clodronate liposomes (to deplete microglia/monocyte-derived macrophages) or control liposomes were co-injected. Magnetic resonance imaging (MRI) was used to evaluate ventricular volume and intraventricular T2* lesion volume (estimating hematoma volume). The brains were harvested after 4 or 72 h for histology to evaluate phagocytosis. Results In adult male rats, CD47 blocking antibody alleviated hydrocephalus development by day 3. In addition, the CD47 blocking antibody reduced intraventricular T2* lesion and T2* non-hypointense lesion size after IVH through day 1 to day 3. Erythrophagocytosis was observed as soon as 4 h after IVH and was enhanced on day 3. Furthermore, intra-hematoma infiltration of CD68, heme oxygenase-1 and ferritin positive phagocytes were upregulated by CD47 blockade by day 3. Clodronate liposomes co-injection caused more severe hydrocephalus and weight loss. Conclusion Blocking CD47 in the hematoma accelerated hematoma clearance and alleviated hemolysis and hydrocephalus development after IVH, suggesting CD47 might be valuable in the future treatment for IVH.
Asunto(s)
Anticuerpos Monoclonales/administración & dosificación , Antígeno CD47/antagonistas & inhibidores , Antígeno CD47/metabolismo , Hemorragia Cerebral Intraventricular/metabolismo , Hematoma/metabolismo , Hidrocefalia/metabolismo , Animales , Hemorragia Cerebral Intraventricular/diagnóstico por imagen , Hemorragia Cerebral Intraventricular/tratamiento farmacológico , Hematoma/diagnóstico por imagen , Hematoma/tratamiento farmacológico , Hidrocefalia/diagnóstico por imagen , Hidrocefalia/tratamiento farmacológico , Imagen por Resonancia Magnética/métodos , Masculino , Ratas , Ratas Endogámicas F344RESUMEN
Intraventricular haemorrhage (IVH) continues to be a major complication of prematurity that can result in cerebral palsy and cognitive impairment in survivors. No optimal therapy exists to prevent IVH or to treat its consequences. IVH varies in severity and can present as a bleed confined to the germinal matrix, small-to-large IVH or periventricular haemorrhagic infarction. Moderate-to-severe haemorrhage dilates the ventricle and damages the periventricular white matter. This white matter injury results from a constellation of blood-induced pathological reactions, including oxidative stress, glutamate excitotoxicity, inflammation, perturbed signalling pathways and remodelling of the extracellular matrix. Potential therapies for IVH are currently undergoing investigation in preclinical models and evidence from clinical trials suggests that stem cell treatment and/or endoscopic removal of clots from the cerebral ventricles could transform the outcome of infants with IVH. This Review presents an integrated view of new insights into the mechanisms underlying white matter injury in premature infants with IVH and highlights the importance of early detection of disability and immediate intervention in optimizing the outcomes of IVH survivors.
Asunto(s)
Hemorragia Cerebral Intraventricular/complicaciones , Hemorragia Cerebral Intraventricular/terapia , Enfermedades del Prematuro , Leucoencefalopatías/etiología , Leucoencefalopatías/terapia , Animales , Hemorragia Cerebral Intraventricular/metabolismo , Hemorragia Cerebral Intraventricular/patología , Humanos , Recién Nacido , Recien Nacido Prematuro , Leucoencefalopatías/metabolismo , Leucoencefalopatías/patologíaRESUMEN
The germinal matrix-intraventricular hemorrhage (GM-IVH) is one of the most important complications of the preterm newborn. Since these children are born at a critical time in brain development, they can develop short and long term neurological, sensory, cognitive and motor disabilities depending on the severity of the GM-IVH. In addition, hemorrhage triggers a microglia-mediated inflammatory response that damages the tissue adjacent to the injury. Nevertheless, a neuroprotective and neuroreparative role of the microglia has also been described, suggesting that neonatal microglia may have unique functions. While the implication of the inflammatory process in GM-IVH is well established, the difficulty to access a very delicate population has lead to the development of animal models that resemble the pathological features of GM-IVH. Genetically modified models and lesions induced by local administration of glycerol, collagenase or blood have been used to study associated inflammatory mechanisms as well as therapeutic targets. In the present study we review the GM-IVH complications, with special interest in inflammatory response and the role of microglia, both in patients and animal models, and we analyze specific proteins and cytokines that are currently under study as feasible predictors of GM-IVH evolution and prognosis.
Asunto(s)
Hemorragia Cerebral Intraventricular/inmunología , Hemorragia Cerebral Intraventricular/metabolismo , Nacimiento Prematuro/fisiopatología , Animales , Encéfalo/metabolismo , Hemorragia Cerebral/metabolismo , Hemorragia Cerebral Intraventricular/fisiopatología , Circulación Cerebrovascular/fisiología , Modelos Animales de Enfermedad , Edad Gestacional , Humanos , Lactante , Recién Nacido , Enfermedades del Recién Nacido , Recien Nacido Prematuro , Enfermedades del Prematuro , Microglía/metabolismo , Microglía/fisiología , Nacimiento Prematuro/inmunologíaRESUMEN
Peri-intraventricular hemorrhage (PIVH) is a common and serious prematurity-related complication in neonates. Adrenocorticotropic hormone (ACTH) has neuroprotective actions and is a candidate to ameliorate brain damage following PIVH. Here, we tested the efficacy of ACTH1-24 on a collagenase-induced lesion of the germinal matrix (GM) in newborn male rats. Animals received microinjection of the vehicle (PBS, 2 µl) or collagenase type VII (0.3 IU) into the GM/periventricular tissue on postnatal day (PN) 2. Twelve hours later pups received microinjection of either the agonist ACTH1-24 (0.048 mg/kg), or the antagonist SHU9119 (antagonist of MCR3/MCR4 receptors, 0.01 mg/kg), or their combination. Morphological outcomes included striatal injury extension, neuronal and glial cells counting, and immunohistochemical expression of brain lesion biomarkers ipsilateral and contralateral to the hemorrhagic site. Data were evaluated on PN 8. Collagenase induced PIVH and severe ipsilateral striatal lesion. ACTH1-24 dampened the deleterious effects of collagenase-induced hemorrhage in significantly reducing the extension of the damaged area, the striatal neuronal and glial losses, and the immunoreactive expression of the GFAP, S100ß, and NG2-glia biomarkers in the affected periventricular area. SHU9119 blocked the glial density rescuing effect of ACTH1-24. ACTH1-24 could be further evaluated to determine its suitability for preclinical models of PVH in premature infants.
Asunto(s)
Hormona Adrenocorticotrópica/metabolismo , Encéfalo/patología , Hemorragia Cerebral Intraventricular/metabolismo , Neuroglía/fisiología , Neuronas/fisiología , Fármacos Neuroprotectores/metabolismo , Péptidos/metabolismo , Nacimiento Prematuro/metabolismo , Animales , Animales Recién Nacidos , Antígenos/metabolismo , Colagenasas/metabolismo , Proteína Ácida Fibrilar de la Glía/metabolismo , Humanos , Masculino , Proteoglicanos/metabolismo , Ratas , Ratas Wistar , Subunidad beta de la Proteína de Unión al Calcio S100/metabolismoRESUMEN
Background and Purpose- Our recent study demonstrated that release of Prx2 (peroxiredoxin 2) from red blood cells (RBCs) is involved in the inflammatory response and brain injury after intracerebral hemorrhage. The current study investigated the role of extracellular Prx2 in hydrocephalus development after experimental intraventricular hemorrhage. Methods- There were 4 parts in this study. First, Sprague-Dawley rats received an intraventricular injection of lysed RBC or saline and were euthanized at 1 hour for Prx2 measurements. Second, rats received an intraventricular injection of Prx2, deactivated Prx2, or saline. Third, lysed RBC was coinjected with conoidin A, a Prx2 inhibitor, or vehicle. Fourth, rats received Prx2 injection and were treated with minocycline or saline (i.p.). The effects of Prx2 and the inhibitors were examined using magnetic resonance imaging assessing ventriculomegaly, histology assessing ventricular wall damage, and immunohistochemistry to assess inflammation, particularly at the choroid plexus. Results- Intraventricular injection of lysed RBC resulted in increased brain Prx2 and hydrocephalus. Intraventricular injection of Prx2 alone caused hydrocephalus, ventricular wall damage, activation of choroid plexus epiplexus cells (macrophages), and an accumulation of neutrophils. Conoidin A attenuated lysed RBC-induced injury. Systemic minocycline treatment reduced the epiplexus cell activation and hydrocephalus induced by Prx2. Conclusions- Prx2 contributed to the intraventricular hemorrhage-induced hydrocephalus, probably by inducing inflammatory responses in choroid plexus and ventricular wall damage.
Asunto(s)
Hemorragia Cerebral Intraventricular/metabolismo , Plexo Coroideo/metabolismo , Hidrocefalia/metabolismo , Inflamación/metabolismo , Macrófagos/metabolismo , Peroxirredoxinas/metabolismo , Animales , Antiinflamatorios/farmacología , Hemorragia Cerebral Intraventricular/complicaciones , Plexo Coroideo/efectos de los fármacos , Plexo Coroideo/patología , Modelos Animales de Enfermedad , Epéndimo/efectos de los fármacos , Epéndimo/patología , Femenino , Hidrocefalia/etiología , Hylobatidae , Inflamación/patología , Inyecciones Intraventriculares , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/patología , Masculino , Minociclina/farmacología , Neutrófilos/efectos de los fármacos , Neutrófilos/patología , Peroxirredoxinas/antagonistas & inhibidores , Peroxirredoxinas/farmacología , Quinoxalinas/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
Intraventricular hemorrhage (IVH) is a frequent complication of prematurity that is associated with high neonatal mortality and morbidity. IVH is accompanied by red blood cell (RBC) lysis, hemoglobin (Hb) oxidation, and sterile inflammation. Here we investigated whether extracellular Hb, metHb, ferrylHb, and heme contribute to the inflammatory response after IVH. We collected cerebrospinal fluid (CSF) (n = 20) from premature infants with grade III IVH at different time points after the onset of IVH. Levels of Hb, metHb, total heme, and free heme were the highest in CSF samples obtained between days 0 and 20 after the onset of IVH and were mostly non-detectable in CSF collected between days 41 and 60 of post-IVH. Besides Hb monomers, we detected cross-linked Hb dimers and tetramers in post-IVH CSF samples obtained in days 0-20 and 21-40, but only Hb tetramers were present in CSF samples obtained after 41-60 days. Vascular cell adhesion molecule-1 (VCAM-1) and interleukin-8 (IL-8) levels were higher in CSF samples obtained between days 0 and 20 than in CSF collected between days 41 and 60 of post-IVH. Concentrations of VCAM-1, intercellular adhesion molecule-1 (ICAM-1), and IL-8 strongly correlated with total heme levels in CSF. Applying the identified heme sources on human brain microvascular endothelial cells revealed that Hb oxidation products and free heme contribute to the inflammatory response. We concluded that RBC lysis, Hb oxidation, and heme release are important components of the inflammatory response in IVH. Pharmacological interventions targeting cell-free Hb, Hb oxidation products, and free heme could have potential to limit the neuroinflammatory response following IVH.
Asunto(s)
Encéfalo/patología , Hemorragia Cerebral Intraventricular/metabolismo , Células Endoteliales/metabolismo , Eritrocitos/patología , Hemo/líquido cefalorraquídeo , Hemoglobinas/líquido cefalorraquídeo , Inflamación/metabolismo , Nacimiento Prematuro/metabolismo , Femenino , Humanos , Recién Nacido , Recien Nacido Prematuro , Molécula 1 de Adhesión Intercelular/líquido cefalorraquídeo , Interleucina-8/líquido cefalorraquídeo , Masculino , Inflamación Neurogénica , Oxidación-Reducción , Nacimiento Prematuro/inmunología , Molécula 1 de Adhesión Celular Vascular/líquido cefalorraquídeoRESUMEN
The lysis of red blood cells was shown to occur in human ruptured atherosclerotic lesions and intraventricular hemorrhage (IVH) of the brain. Liberated cell-free hemoglobin was found to undergo oxidation in both pathologies. We hypothesize that hemoglobin-derived peptides are generated during hemoglobin oxidation both in complicated atherosclerotic lesions and IVH of the brain, triggering endothelial cell dysfunction. Oxidized hemoglobin and its products were followed with spectrophotometry, LC-MS/MS analysis and detection of the cross-linking of globin chains in complicated atherosclerotic lesions of the human carotid artery and the hemorrhaged cerebrospinal liquid of preterm infants. The vascular pathophysiologic role of oxidized hemoglobin and the resultant peptides was assessed by measuring endothelial integrity, the activation of endothelial cells and the induction of proinflammatory genes. Peptide fragments of hemoglobin (VNVDEVGGEALGRLLVVYPWTQR, LLVVYPWTQR, MFLSFPTTK, VGAHAGEYGAELERMFLSFPTTK, and FLASVSTVLTSKYR) were identified in ruptured atherosclerotic lesions and in IVH of the human brain. Fragments resulting from the oxidation of hemoglobin were accompanied by the accumulation of ferryl hemoglobin. Similar to complicated atherosclerotic lesions of the human carotid artery, a high level of oxidized and cross-linked hemoglobin was observed in the cerebrospinal fluid after IVH. Haptoglobin inhibited hemoglobin fragmentation provoked by peroxide. The resultant peptides failed to bind haptoglobin or albumin. Peptides derived from hemoglobin oxidation and ferryl hemoglobin induced intercellular gap formation, decreased junctional resistance in the endothelium, and enhanced monocyte adhesion to endothelial cells. Enhanced expression of TNF and the activation of NLRP3 and CASP1 followed by the increased generation of IL-1ß and nuclear translocation of the NF-κß transcription factor occurred in response to hemoglobin-derived peptides, and ferryl hemoglobin in endothelium was upregulated in both pathologies. We conclude that the oxidation of hemoglobin in complicated atherosclerotic lesions and intraventricular hemorrhage of the brain generates peptide fragments and ferryl hemoglobin with the potential to trigger endothelial cell dysfunction.
Asunto(s)
Enfermedades de las Arterias Carótidas/metabolismo , Hemorragia Cerebral Intraventricular/metabolismo , Endotelio Vascular/fisiopatología , Hemoglobinas , Encéfalo/metabolismo , Encéfalo/patología , Enfermedades de las Arterias Carótidas/patología , Células Cultivadas , Hemorragia Cerebral Intraventricular/patología , Cromatografía Liquida , Hemoglobinas/química , Hemoglobinas/metabolismo , Células Endoteliales de la Vena Umbilical Humana , Humanos , Oxidación-Reducción , Fragmentos de Péptidos/química , Fragmentos de Péptidos/metabolismo , Espectrometría de Masas en TándemRESUMEN
Background: The discovery of intracranial lymphatic system provides a new theory about cerebrospinal fluid circulation. In order to investigate the role of lymphatic drainage in hydrocephalus after intraventricular hemorrhage (IVH), three parts of experiments were performed.Methods: IVH model was produced by 200ul autologous blood injection into lateral ventricle of SD male rats. Perls Prussian blue reaction was used to confirm the ability of iron drainage through deep cervical lymph nodes (DCLNs) in part I. The part II measured the volume of ventricles by MR scanner , and the part III compared the ferritin level of area surrounding ventricles by Western blotting after IVH.Results: The ability of iron drainage through DCLNs was confirmed. DCLNs-excised rats got a 19.6% higher incidence of hydrocephalus than DCLNs-preserved rats at day 28 after IVH. And DCLNs-excised group showed higher ferritin level than DCLNs-preserved group at day 3 after IVH.Conclusion: The whole experiments revealed the role of intracranial lymphatic system in discharging iron out of central nervous system at acute phase of IVH, and proved the function in alleviating and avoiding hydrocephalus at chronic phase after IVH.
Asunto(s)
Hemorragia Cerebral Intraventricular/metabolismo , Ventrículos Cerebrales/metabolismo , Ferritinas/metabolismo , Hidrocefalia/metabolismo , Hierro/metabolismo , Sistema Linfático/metabolismo , Animales , Western Blotting , Hemorragia Cerebral Intraventricular/complicaciones , Modelos Animales de Enfermedad , Hidrocefalia/etiología , Linfa/metabolismo , Imagen por Resonancia Magnética , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Severe intraventricular hemorrhage (IVH) in premature infants triggers reactive gliosis, causing acute neuronal death and glial scar formation. Transplantation of mesenchymal stem cells (MSCs) has often showed improved CNS recovery in an IVH model, but whether this response is related to reactive glial cells is still unclear. Herein, we suggest that MSCs impede the response of reactive microglia rather than astrocytes, thereby blocking neuronal damage. Astrocytes alone showed mild reactiveness under hemorrhagic conditions mimicked by thrombin treatment, and this was not blocked by MSC-conditioned medium (MSC-CM) in vitro. In contrast, thrombin-induced microglial activation and release of proinflammatory cytokines were inhibited by MSC-CM. Interestingly, astrocytes showed greater reactive response when co-cultured with microglia, and this was abolished in the presence of MSC-CM. Gene expression profiles in microglia revealed that transcript levels of genes for immune response and proinflammatory cytokines were altered by thrombin treatment. This result coincided with the robust phosphorylation of STAT1 and p38 MAPK, which might be responsible for the production and release of proinflammatory cytokines. Furthermore, application of MSC-CM diminished thrombin-mediated phosphorylation of STAT1 and p38 MAPK, supporting the acute anti-inflammatory role of MSCs under hemorrhagic conditions. In line with this, activation of microglia and consequent cytokine release were impaired in Stat1-null mice. However, reactive response in Stat1-deficient astrocytes was maintained. Taken together, our results demonstrate that MSCs mainly block the activation of microglia involving STAT1-mediated cytokine release and subsequent reduction of reactive astrocytes.
Asunto(s)
Astrocitos/metabolismo , Hemorragia Cerebral Intraventricular/metabolismo , Modelos Animales de Enfermedad , Células Madre Mesenquimatosas/metabolismo , Microglía/metabolismo , Animales , Animales Recién Nacidos , Astrocitos/patología , Células Cultivadas , Hemorragia Cerebral Intraventricular/terapia , Mediadores de Inflamación/antagonistas & inhibidores , Mediadores de Inflamación/metabolismo , Masculino , Trasplante de Células Madre Mesenquimatosas/métodos , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microglía/patología , Ratas , Ratas Sprague-DawleyRESUMEN
The brain is one of the main organs affected by hypoxia and reoxygenation in the neonatal period and one of the most vulnerable to oxidative stress. Hypoxia/ischemia and reoxygenation leads to impairment of neurogenesis, disruption of cortical migration, mitochondrial damage and neuroinflammation. The extent of the injury depends on the clinical manifestation in the affected regions. Preterm newborns are highly vulnerable, and they exhibit severe clinical manifestations such as intraventricular hemorrhage (IVH), retinopathy of prematurity (ROP) and diffuse white matter injury (DWMI) among others. In the neonatal period, the accumulation of high levels of reactive oxygen species exacerbated by the immature antioxidant defense systems in represents cellular threats that, if they exceed or bypass physiological counteracting mechanisms, are responsible of significant neuronal damage. Several experimental models in mice mimic the consequences of perinatal asphyxia and the use of oxygen in the reanimation process that produce brain injury. The aim of this review is to highlight brain damage associated with oxidative stress in different murine models of hypoxia/ischemia and reoxygenation.
Asunto(s)
Asfixia Neonatal/patología , Isquemia Encefálica/patología , Encéfalo/patología , Hemorragia Cerebral Intraventricular/patología , Hipoxia/patología , Retinopatía de la Prematuridad/patología , Animales , Asfixia Neonatal/metabolismo , Encéfalo/metabolismo , Isquemia Encefálica/metabolismo , Hemorragia Cerebral Intraventricular/metabolismo , Modelos Animales de Enfermedad , Humanos , Hipoxia/metabolismo , Recién Nacido , Ratones , Oxidación-Reducción , Estrés Oxidativo , Oxígeno/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Retinopatía de la Prematuridad/metabolismoRESUMEN
BACKGROUND: To investigate the clinical values of serum melatonin and αII spectrin cleavage products (SBDPs) in assessing the severity of brain injury in preterm infants. METHODS: Sixty-four premature infants in total were selected and classified into the brain injury group (BI, n = 30) and the non-brain injury group (CON, n = 34) according to cranial imaging examination. The serum melatonin and SBDPs were detected by ELISA. All the preterm infants were received NBNA testing at 40 weeks of corrected gestational age. RESULTS: The levels of melatonin and SBDPs in the BI group were significantly higher than the CON group (p < 0.05) and the levels in the infants with severe brain injury were significantly higher than those with mild brain injury (p < 0.05), as well as exhibiting a negative correlation with the NBNA score at 40 weeks of corrected gestational age (p < 0.05). CONCLUSIONS: Detecting melatonin and SBDPs has clinical value in diagnosing and assessing the severity of brain injury in preterm infants.
Asunto(s)
Encéfalo/diagnóstico por imagen , Infarto Cerebral/sangre , Hemorragia Cerebral Intraventricular/sangre , Hemorragias Intracraneales/sangre , Leucomalacia Periventricular/sangre , Melatonina/sangre , Espectrina/sangre , Lesiones Encefálicas/sangre , Lesiones Encefálicas/diagnóstico por imagen , Lesiones Encefálicas/metabolismo , Lesiones Encefálicas/fisiopatología , Estudios de Casos y Controles , Infarto Cerebral/diagnóstico por imagen , Infarto Cerebral/metabolismo , Infarto Cerebral/fisiopatología , Hemorragia Cerebral Intraventricular/diagnóstico por imagen , Hemorragia Cerebral Intraventricular/metabolismo , Femenino , Humanos , Recién Nacido , Recien Nacido Prematuro , Enfermedades del Prematuro/sangre , Enfermedades del Prematuro/diagnóstico por imagen , Enfermedades del Prematuro/metabolismo , Enfermedades del Prematuro/fisiopatología , Hemorragias Intracraneales/diagnóstico por imagen , Hemorragias Intracraneales/metabolismo , Hemorragias Intracraneales/fisiopatología , Leucomalacia Periventricular/diagnóstico por imagen , Leucomalacia Periventricular/metabolismo , Leucomalacia Periventricular/fisiopatología , Imagen por Resonancia Magnética , Masculino , Índice de Severidad de la Enfermedad , Espectrina/metabolismoRESUMEN
Intraventricular hemorrhage (IVH) is a common complication of prematurity in infants born at 23-28 weeks of gestation. Survivors exhibit impaired growth of the cerebral cortex and neurodevelopmental sequeale, but the underlying mechanism(s) are obscure. Previously, we have shown that neocortical neurogenesis continues until at least 28 gestational weeks. This renders the prematurely born infants vulnerable to impaired neurogenesis. Here, we hypothesized that neurogenesis is impaired by IVH, and that signaling through GSK3ß, a critical intracellular kinase regulated by Wnt and other pathways, mediates this effect. These hypotheses were tested observationally in autopsy specimens from premature infants, and experimentally in a premature rabbit IVH model. Significantly, in premature infants with IVH, the number of neurogenic cortical progenitor cells was reduced compared with infants without IVH, indicating acutely decreased neurogenesis. This finding was corroborated in the rabbit IVH model, which further demonstrated reduction of upper layer cortical neurons after longer survival. Both the acute reduction of neurogenic progenitors, and the subsequent decrease of upper layer neurons, were rescued by treatment with AR-A014418, a specific inhibitor of GSK3ß. Together, these results indicate that IVH impairs late stages of cortical neurogenesis, and suggest that treatment with GSK3ß inhibitors may enhance neurodevelopment in premature infants with IVH.
Asunto(s)
Apoptosis/efectos de los fármacos , Hemorragia Cerebral Intraventricular/metabolismo , Glucógeno Sintasa Quinasa 3 beta/antagonistas & inhibidores , Células-Madre Neurales/efectos de los fármacos , Neurogénesis/efectos de los fármacos , Tiazoles/farmacología , Urea/análogos & derivados , Animales , Western Blotting , Estudios de Casos y Controles , Recuento de Células , Proliferación Celular , Corteza Cerebral , Hemorragia Cerebral Intraventricular/patología , Modelos Animales de Enfermedad , Células Ependimogliales/efectos de los fármacos , Células Ependimogliales/metabolismo , Humanos , Inmunohistoquímica , Recien Nacido Extremadamente Prematuro , Recién Nacido , Antígeno Ki-67/metabolismo , Ventrículos Laterales , Células-Madre Neurales/metabolismo , Células-Madre Neurales/patología , Factor de Transcripción PAX6/metabolismo , Fosforilación , Células Piramidales/efectos de los fármacos , Células Piramidales/metabolismo , Células Piramidales/patología , Conejos , Reacción en Cadena en Tiempo Real de la Polimerasa , Proteína de Retinoblastoma/metabolismo , Factores de Transcripción SOXB1/metabolismo , Proteínas de Dominio T Box/metabolismo , Urea/farmacología , Sustancia BlancaRESUMEN
Germinal matrix hemorrhage-intraventricular hemorrhage (GMH-IVH) remains a serious complication in the preterm newborn. The significant increase of survival rates in extremelye preterm newborns has also contributed to increase the absolute number of patients developing GMH-IVH. However, there are relatively few available animal models to understand the underlying mechanisms and peripheral markers or prognostic tools. In order to further characterize central complications and evolution of GMH-IVH, we injected collagenase intraventricularly to P7 CD1 mice and assessed them in the short (P14) and the long term (P70). Early complications at P14 included ventricle enlargement, increased bleeding, and inflammation. These alterations were maintained at P70, when increased tau phosphorylation and decreased neurogenesis were also observed, resulting in impaired learning and memory in these early adult mice. We additionally analyzed peripheral blood biomarkers in both our mouse model and preterm newborns with GMH-IVH. While MMP9 levels were not significantly altered in mice or newborns, reduced gelsolin levels and increased ubiquitin carboxy-terminal hydrolase L1 and tau levels were detected in GMH-IVH patients at birth. A similar profile was observed in our mouse model after hemorrhage. Interestingly, early changes in gelsolin and carboxy-terminal hydrolase L1 levels significantly correlated with the hemorrhage grade in newborns. Altogether, our data support the utility of this animal model to reproduce the central complications and peripheral changes observed in the clinic, and support the consideration of gelsolin, carboxy-terminal hydrolase L1, and tau as feasible biomarkers to predict the development of GMH-IVH.
Asunto(s)
Encéfalo/patología , Hemorragia Cerebral Intraventricular/psicología , Disfunción Cognitiva/psicología , Aprendizaje/fisiología , Memoria/fisiología , Destreza Motora/fisiología , Animales , Biomarcadores/sangre , Encéfalo/metabolismo , Hemorragia Cerebral Intraventricular/metabolismo , Hemorragia Cerebral Intraventricular/patología , Disfunción Cognitiva/metabolismo , Disfunción Cognitiva/patología , Modelos Animales de Enfermedad , Femenino , Inflamación/metabolismo , Inflamación/patología , Inflamación/psicología , Masculino , Ratones , Fosforilación , Proteínas tau/metabolismoRESUMEN
Mesenchymal stem cell (MSC) transplantation protects against neonatal severe intraventricular hemorrhage (IVH)-induced brain injury by a paracrine rather than regenerative mechanism; however, the paracrine factors involved and their roles have not yet been delineated. This study aimed to identify the paracrine mediator(s) and to determine their role in mediating the therapeutic effects of MSCs in severe IVH. We first identified significant upregulation of brain-derived neurotrophic factor (BDNF) in MSCs compared with fibroblasts, in both DNA and antibody microarrays, after thrombin exposure. We then knocked down BDNF in MSCs by transfection with small interfering (si)RNA specific for human BDNF. The therapeutic effects of MSCs with or without BDNF knockdown were evaluated in vitro in rat neuronal cells challenged with thrombin, and in vivo in newborn Sprague-Dawley rats by injecting 200 µl of blood on postnatal day 4 (P4), and transplanting MSCs (1 × 105 cells) intraventricularly on P6. siRNA-induced BDNF knockdown abolished the in vitro benefits of MSCs on thrombin-induced neuronal cell death. BDNF knockdown also abolished the in vivo protective effects against severe IVH-induced brain injuries such as the attenuation of posthemorrhagic hydrocephalus, impaired behavioral test performance, increased astrogliosis, increased number of TUNEL cells, ED-1+ cells, and inflammatory cytokines, and reduced myelin basic protein expression. Our data indicate that BDNF secreted by transplanted MSCs is one of the critical paracrine factors that play a seminal role in attenuating severe IVH-induced brain injuries in newborn rats.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/metabolismo , Hemorragia Cerebral Intraventricular/metabolismo , Hemorragia Cerebral Intraventricular/terapia , Animales , Factor Neurotrófico Derivado del Encéfalo/genética , Muerte Celular/efectos de los fármacos , Células Cultivadas , Humanos , Hidrocefalia/metabolismo , Etiquetado Corte-Fin in Situ , Recién Nacido , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/citología , Proteína Básica de Mielina/metabolismo , ARN Interferente Pequeño/genética , Ratas , Ratas Sprague-Dawley , Trombina/farmacologíaRESUMEN
Low-grade germinal matrix-intraventricular hemorrhage (GM-IVH) is the most common complication in extremely premature neonates. The occurrence of GM-IVH is highly associated with hemodynamic instability in the premature brain, yet the long-term impact of low-grade GM-IVH on cerebral blood flow and neuronal health have not been fully investigated. We used an innovative combination of frequency-domain near infrared spectroscopy and diffuse correlation spectroscopy (FDNIRS-DCS) to measure cerebral oxygen saturation (SO2) and an index of cerebral blood flow (CBFi) at the infant's bedside and compute an index of cerebral oxygen metabolism (CMRO2i). We enrolled twenty extremely low gestational age (ELGA) neonates (seven with low-grade GM-IVH) and monitored them weekly until they reached full-term equivalent age. During their hospital stay, we observed consistently lower CBFi and CMRO2i in ELGA neonates with low-grade GM-IVH compared to neonates without hemorrhages. Furthermore, lower CBFi and CMRO2i in the former group persists even after the resolution of the hemorrhage. In contrast, SO2 does not differ between groups. Thus, CBFi and CMRO2i may have better sensitivity than SO2 in detecting GM-IVH-related effects on infant brain development. FDNIRS-DCS methods may have clinical benefit for monitoring the evolution of GM-IVH, evaluating treatment response, and potentially predicting neurodevelopmental outcome.