RESUMO
Ischemic stroke is the main reason of disability and mortality in many countries, and currently has limited treatments. The post-stroke inflammation characterized with microglia activation and polarization has been regarded as a promising therapeutic target for ischemic stroke. After ischemia, the activated microglia polarize to classical (M1) phenotype or alternative (M2) phenotype and exhibit biphasic function. Promoting microglia phenotype shift from deleterious M1 phenotype to neuroprotective M2 phenotype will be promising in stroke treatment. Increasing evidence indicates that the erythropoietin-producing human hepatocellular (Eph) receptor A4 (EphA4), a kind of abundant Eph receptor, distributes mainly in neuron and participates in multiple links of pathological changes after ischemia. This paper discussed the hypothesis that EphA4 receptor could affect ischemic brain injury through EphA4/ephrin bidirectional signaling between neuron and microglia, and then explored its underlying mechanisms. We manipulated EphA4/ephrin signaling with either EphA4 overexpression lentiviral vectors or the short hairpin RNA (shRNA) to upregulate or knock down neuronal EphA4 expression. NF-κB inhibitor pyrrolidine dithiocarbamate ammonium salt (PDTC) was applied to block NF-κB pathway. According to the experimental results, upregulated neuronal EphA4 induced by ischemia deteriorated neurological function as well as brain damage by shifting microglia M1-polarization via promoting NF-κB signaling.
Assuntos
Transdiferenciação Celular/efeitos dos fármacos , Dipiridamol/farmacologia , Fibroblastos/metabolismo , Miofibroblastos/metabolismo , Inibidores de Fosfodiesterase/farmacologia , Transdução de Sinais , Actinas/genética , Actinas/metabolismo , Animais , Derivados de Benzeno/farmacologia , Biomarcadores/metabolismo , Linhagem Celular , Transdiferenciação Celular/genética , AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Proteínas Quinases Dependentes de AMP Cíclico/genética , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica , Fatores de Troca do Nucleotídeo Guanina/antagonistas & inibidores , Fatores de Troca do Nucleotídeo Guanina/genética , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Isoquinolinas/farmacologia , Rim/citologia , Rim/metabolismo , Miofibroblastos/citologia , Miofibroblastos/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Ratos , Sulfonamidas/farmacologia , Sulfonas/farmacologia , Fator de Crescimento Transformador beta1/farmacologiaRESUMO
Accumulating evidence indicates that post-injury inflammation characterized by activated microglia contributes much to the neuropathology of ischemic injury. Several studies have demonstrated that microglia exhibit two entirely different functional activation states, referred to as classically activated (M1) and alternatively activated (M2) phenotype. Promoting microglial phenotype to switch from M1 dominant to M2 dominant might be a promising approach for handling ischemic injury. However, the comprehensive mechanism that underlines microglia polarization in ischemic brain remains unclear. Neuronal erythropoietin-producing human hepatocellular carcinoma cell receptor 4 (EphA4), the richest Eph receptor in the central nervous system (CNS), upregulate after ischemia and may have the potential to regulate microglia activation. We hypothesized that modulating EphA4/ephrin signaling could affect ischemic injury through controlling microglia polarization. We therefore knocked down neuronal EphA4 with short hairpin RNA (shRNA) and determined the role of EphA4/ephrin signaling in oxygen-glucose deprivation and reperfusion (OGD/R)-induced injury. We found that EphA4 shRNA treatment attenuated OGD/R-induced apoptosis and microglia proliferation. Neuronal EphA4 knockdown also promoted microglial M2 polarization, which reduced pro-inflammatory mediators and released anti-inflammatory cytokines as well as neurotrophic factors. We further revealed that EphA4 shRNA treatment functioned through RhoA/Rho-associated kinase 2 (ROCK2) signaling, a key mediator of microglia alternative activation. Together, these data suggested that blockage of EphA4/ephrin signaling between neuron and microglia decreased OGD/R-induced injury by promoting alternative activation of microglia via RhoA/ROCK2 signaling.
Assuntos
Apoptose , Microglia/metabolismo , Receptor EphA4/fisiologia , Traumatismo por Reperfusão/patologia , Células Cultivadas , Humanos , Fenótipo , Traumatismo por Reperfusão/etiologia , Transdução de Sinais , Quinases Associadas a rho/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismoRESUMO
Multiple cellular components are involved in the complex pathological process following central nervous system (CNS) injury, including neurons, glial cells and endothelial cells. Previous studies and neurotherapeutic clinical trials have assessed the molecular mechanisms that underlie neuronal cell death following CNS injury. However, this approach has largely failed to reduce CNS damage or improve the functional recovery of patients. Erythropoietin-producing human hepatocellular (Eph) receptors and ephrin ligands have attracted considerable attention since their discovery, due to their extensive distribution and unique bidirectional signaling between astrocytes and neurons. Previous studies have investigated the roles of Eph/ephrin bidirectional signaling in the developing central nervous system. It was determined that Eph/ephrin bidirectional signaling is expressed in various CNS regions and cell types, and that it serves diverse roles in the adult CNS. In the present review, the roles of Eph/ephrin bidirectional signaling in CNS injuries are assessed.
RESUMO
Stress-induced inflammatory responses in the portal system are characterized by elevations in serum concentrations of interleukin-6 (IL-6) and endotoxins such as lipopolysaccharides (LPS). LPS translocation from the intestinal to the capillary lumen occurs via LPS endocytosis by the capillary endothelium. Because the capillary endothelium of the small intestinal submucosa is fenestrated, we determined the role of pore modifications within the fenestrated endothelium in relaying inflammatory stress responses in the portal vein. We evaluated changes in the diameter and density of endothelial pores of the lamina propria of intestinal villi induced by continuous light (CL) exposure for 48 h and the correlation between these changes and serum IL-6 concentration in the portal vein in a rat model. We found significant increases in both the pore diameter and density, accompanied by a significant increase in portal IL-6 concentration; these changes were significantly attenuated by pretreatment with propranolol, a beta adrenergic receptor antagonist. In contrast, intravenous noradrenaline administration mimicked CL-induced modifications of the diameter and density of pores and the elevation of portal vein IL-6 concentration. These findings suggested that stress-induced inflammatory responses in the portal system may be a part of the modifications of the endothelial pores triggered by sympathetic activation.
Assuntos
Capilares/patologia , Capilares/ultraestrutura , Endotélio Vascular/patologia , Endotélio Vascular/ultraestrutura , Estresse Psicológico/patologia , Animais , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Mucosa Intestinal/irrigação sanguínea , Masculino , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Ratos , Ratos WistarRESUMO
Histidine decarboxylase (HDC) catalyzes the biosynthesis of histamine from L-histidine and is expressed throughout the mammalian nervous system by histaminergic neurons. Histaminergic neurons arise in the posterior mesencephalon during the early embryonic period and gradually develop into two histaminergic substreams around the lateral area of the posterior hypothalamus and the more anterior peri-cerebral aqueduct area before finally forming an adult-like pattern comprising five neuronal clusters, E1, E2, E3, E4, and E5, at the postnatal stage. This distribution of histaminergic neuronal clusters in the rat hypothalamus appears to be a consequence of neuronal development and reflects the functional differentiation within each neuronal cluster. However, the close linkage between the locations of histaminergic neuronal clusters and their physiological functions has yet to be fully elucidated because of the sparse information regarding the location and orientation of each histaminergic neuronal clusters in the hypothalamus of rats and mice. To clarify the distribution of the five-histaminergic neuronal clusters more clearly, we performed an immunohistochemical study using the anti-HDC antibody on serial sections of the rat hypothalamus according to the brain maps of rat and mouse. Our results confirmed that the HDC-immunoreactive (HDCi) neuronal clusters in the hypothalamus of rats and mice are observed in the ventrolateral part of the most posterior hypothalamus (E1), ventrolateral part of the posterior hypothalamus (E2), ventromedial part from the medial to the posterior hypothalamus (E3), periventricular part from the anterior to the medial hypothalamus (E4), and diffusely extended part of the more dorsal and almost entire hypothalamus (E5). The stereological estimation of the total number of HDCi neurons of each clusters revealed the larger amount of the rat than the mouse. The characterization of histaminergic neuronal clusters in the hypothalamus of rats and mice may provide useful information for further investigations.
Assuntos
Histamina/metabolismo , Hipotálamo/citologia , Neurônios/metabolismo , Algoritmos , Animais , Mapeamento Encefálico , Aqueduto do Mesencéfalo/metabolismo , Histidina Descarboxilase/metabolismo , Hipotálamo/crescimento & desenvolvimento , Hipotálamo/metabolismo , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/classificação , Ratos , Ratos Wistar , Terminologia como AssuntoRESUMO
BACKGROUND: Human thoracic brown adipose tissue (BAT), composed of several subdivisions, is a well-known target organ of many clinical studies; however, the functional contribution of each part of human thoracic BAT remains unknown. The present study analyzed the significance of each part of human thoracic BAT in the association between regional distribution, cellularity, and factors involved in the functional regulation of thoracic BAT. METHODS: We analyzed 1550 healthy adults who underwent medical check-ups by positron-emission tomography and computed tomography (PET-CT) imaging, 8 cadavers, and 78 autopsy cases in an observational study. We first characterized the difference between the mediastinum and the supraclavicular areas using counts of BAT detection and conditions based on PET-CT outcomes. The measurable important area was then subjected to systematic anatomical and immunohistochemical analyses using anti-uncoupling protein 1 (UCP1) antibody to characterize the cellularity in association with age and sex. RESULTS: In PET-CT scanning, the main site of thoracic BAT was the mediastinum rather than the supraclavicular area (P < 0.05). Systemic macroanatomy revealed that the thumb-sized BAT in the posterior mediastinal descending para-aortic area (paBAT) had feeding vessels from the posterior intercostal arteries and veins and sympathetic/parasympathetic innervation from trunks of the sympathetic and vagus nerves, respectively. Immunohistochemical analysis indicated that the paBAT exhibited immunoreactivity for tyrosine hydroxylase and vesicular acetylcholine transporter located in the pericellular nervous fibers and intracellular UCP1. The brown adipose cells of paBAT showed age-dependent decreases in UCP1 expression (P < 0.05), accompanied by a significant increase in vacuole formation, indicating fat accumulation (P < 0.05), from 10 to 37 years of age (P < 0.01). CONCLUSIONS: paBAT may be one of the essential sites for clinical application in BAT study because of its visible anatomy with feeding vessels and sympathetic/parasympathetic innervation functionally affected by outer condition and senescence.
Assuntos
Tecido Adiposo Marrom/diagnóstico por imagem , Glomos Para-Aórticos/diagnóstico por imagem , Radiografia Torácica , Tórax/metabolismo , Tecido Adiposo Marrom/citologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Autopsia , Cadáver , Criança , Feminino , Fluordesoxiglucose F18 , Humanos , Canais Iônicos/biossíntese , Masculino , Pessoa de Meia-Idade , Proteínas Mitocondriais/biossíntese , Glomos Para-Aórticos/citologia , Tomografia por Emissão de Pósitrons , Tórax/citologia , Proteína Desacopladora 1 , Vacúolos/metabolismoRESUMO
OBJECTIVE: To study the clinical characteristics and electrophysiological changes in patients with atypical myasthenia gravis (MG). METHODS: The characteristics of the clinical symptoms and electrophysiological changes were investigated in 32 patients with atypical MG diagnosed in our hospital from January 2004 to December 2008. RESULTS: The ages of the patients ranged from 7 to 70 years. Five patients were diagnosed to have ocular MG (OMG), among whom 2 patient only complained of eye discomfort and blurred vision. Twenty-seven patients had generalized MG, and 6 of them showed muscle weakness of the limbs with or without mild difficulty in swallowing or respiratory muscles, but free of muscle dysfunctions in muscles of eyes, head, neck or face. Another 2 patients manifested muscular atrophy. Twenty-three patients (71.9%) displayed both fluctuating symptoms and positive results of fatigue test. Twenty-nine patients (90.6%) have positive results in the neostigmine test. Two patients in the OMG group (40.0%) showed positive results in the low frequency repetitive nerve stimulation (LFRNS), as compared with the 21 patients in the generalized MG group (71.9%) showing positive results. The total positivity rate of LFRNS was 71.9% in the total patients, consistent with the published data. CONCLUSIONS: In MG patients with atypical clinical symptoms, negative results of neostigmine test and fatigue test, LFRNS test can be an indispensable method to increase detection rate of MG and reduce erroneous or missed diagnosis.
Assuntos
Fenômenos Eletrofisiológicos , Miastenia Gravis/fisiopatologia , Adolescente , Adulto , Idoso , Criança , Eletromiografia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Miastenia Gravis/classificação , Miastenia Gravis/diagnóstico , Neostigmina , Estimulação Elétrica Nervosa Transcutânea , Adulto JovemRESUMO
OBJECTIVE: To study the calculation of cerebral blood flow loss ratio after cerebral artery stenosis using the data obtained from transcranial Doppler sonography (TCD), by deducing regional cerebral blood flow loss index (rCBFLI). METHODS AND RESULTS: The data of TCD performed in 31 cases of unilateral stenosis in the middle cerebral artery (MCA) were collected and analyzed for deducing the formula for calculating the MCA stenosis index (STI): STI=1 (Vm(0)/Vm(1)) x (PI(1)/PI(0)), where Vm(0) is the mean normal velocity of MCA blood flow (derived from the measurements in 908 normal subjects at varied ages receiving TCD) and Vm(1) the mean velocity at the stenosed MCA, and PI(0) and PI(1) stand for the pulsatility index before and after stenosis. The ratio of PI(1) to PI(0) is likely to be equivalent to the ratio of Q(1) to Q(0) (Q(0) and Q(1) represent the volume of blood flow in the MCA in normal condition and after stenosis, respectively), thus the equivalence can be extended as rCBFLI=(1 Q(1)/Q(0)) x 100%=(1 PI(1)/PI(0)) x 100%. rCBFLI after MCA stenosis was calculated in 31 cases accordingly, and by means of correlation analysis, we found positive correlation between rCBFLI and Vm(1) (r=0.76, P < 0.001) and between Vm(1) and STI (r=0.85, P < 0.001). In the same way, rCBFLI of 55 sides with MCA stenosis in 43 cases were analyzed, and positive correlation between rCBFLI and Vm (r=0.76, P < 0.001) and between rCBFLI and STI (r=0.83, P < 0.001) was found. CONCLUSION: rCBFLI can be used to evaluate the degree of regional cerebral blood flow loss due to artery stenosis, and the combination of rCBFLI and STI may provide an insight into the changes of cerebral hemodynamics in this pathologic condition.