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Objective: The objective of this study is to investigate the aggressive infiltration of glioblastoma into adjacent brain tissue, considering its challenging prognosis. Initially classified as an intergenic non-coding RNA, we aim to elucidate the functional implications of LINC01138 in glioblastoma. Method: Glioma grading was performed utilizing H&E staining, which unveiled distinct nuclear morphology in high-grade gliomas. The downregulation of LINC01138 in glioma tissues was corroborated through qRT-PCR and gel electrophoresis, concurrently identifying two previously unrecognized LINC01138 isoforms. Expression profiling of all four LINC01138 isoforms was executed in glioma cell lines (A172, SHG-44, U251, U87-MG). The impact of LINC01138 overexpression in U87-MG and U251 cells was evaluated for cell proliferation, migration, and invasion through cell counting, CCK-8 analysis, and Transwell assays. Furthermore, the suppression of LINC01138 in SHG-44 cells substantiated its involvement in fostering tumor malignancy. Transcriptome sequencing revealed the inhibitory influence of LINC01138 on IGF1 expression. These findings contribute to an enriched comprehension of glioma biology by exploring the engagement of LINC01138 through diverse methodologies, thereby elucidating its potential therapeutic significance. Results: Our investigation elucidates the intricate involvement of LINC01138 in gliomas. High-grade gliomas are characterized by elevated cell density and distinctive nuclear features. LINC01138 demonstrates a substantial downregulation in glioma tissues, with the identification of two novel isoforms. The expression of all four LINC01138 isoforms is notably diminished in both glioma tissues and cell lines. Elevated expression of LINC01138 demonstrates inhibitory effects on tumor cell proliferation, migration, and invasion, while its downregulation exacerbates malignancy. The regulatory function of LINC01138 as a repressor of IGF1 expression was elucidated through transcriptome sequencing. Conclusion: The LINC01138 isoforms display notable tumor-suppressive effects, suggesting a promising potential for impeding glioma progression.
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High mobility group box 1 (HMGB1) is a highly conserved and ubiquitous nuclear protein in eukaryotic cells. In response to stress, it transfers from the nucleus to the cytoplasm and finally, to the extracellular matrix, participating in inflammation and carcinogenesis. Increased HMGB1 protein levels are frequently associated with the reduced survival of patients with glioma. HMGB1 plays contextual roles depending on its subcellular localization. However, the mechanisms underlying its subcellular localization and secretion remain unclear. In the present study, the subcellular localization and secretion of HMGB1 in starved glioma cells were investigated using immunofluorescence microscopy, enzymelinked immunosorbent assay, subcellular fractionation, western blotting and immunoelectron microscopy. The results demonstrated that starvation induced HMGB1 translocation from the nucleus to the cytoplasm and finally, to the extracellular milieu in glioma cells. HMGB1 was localized in the mitochondria, endoplasmic reticulum (ER), peroxisomes, autophagosomes, lysosomes, endosomes and the cytoskeleton. Immunoelectron microscopy confirmed that HMGB1 was present within or around cytosolic compartments. Subcellular fractionation further demonstrated that HMGB1 transferred to membranebound compartments. In addition, HMGB1 was localized to specific contact areas between the ER and mitochondria, known as mitochondriaassociated membranes. On the whole, the results of the present study suggest that starvation induces HMGB1 secretion, which can be inhibited through the suppression of autophagy. Starvation insult induces HMGB1 translocation to the cytosolic compartments of glioma cells, and autophagy may be involved in the extracellular secretion of HMGB1 in starved glioma cells.
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Glioma , Proteína HMGB1 , Humanos , Línea Celular , Ensayo de Inmunoadsorción Enzimática , Proteína HMGB1/metabolismo , Lisosomas/metabolismo , Macrófagos/metabolismo , Glioma/metabolismoRESUMEN
RATIONALE AND OBJECTIVES: The classification of patients based on pathology and molecular features is important for improving WHO grade II glioma patient prognosis, especially for the initially diagnosed patients. Less invasive and more convenient methods for the prediction of the pathological type and gene status are desired. MATERIALS AND METHODS: This study investigates the ability to use conventional magnetic resonance imaging (MRI) and computed tomography (CT) features for determining the Isocitrate Dehydrogenase (IDH)-mutant and 1p/19q-codeletion status, through a retrospective review of information obtained from 189 WHO grade II glioma patients. Diffuse astrocytoma (IDH-mutant), Diffuse astrocytoma (IDH- wildtype) and Oligodendroglioma (IDH-mutant and 1p/19q co-deletion) were included in this cohort. All patients were divided into IDH-mutant group and IDH-wildtype group according to the IDH R132H mutation status. Moreover, all patients were divided into 1p/19q co-deletion group and 1p/19q non-codeletion group according to the 1p and 19q chromosome status. Patients underwent pre-operative CT and MRI scans, followed by operation and histopathological analyses, including immunohistochemistry and polymerase chain reaction analysis for IDH mutants, and fluorescence capillary electrophoresis analysis for the 1p/19q co-deletion. The χ2 test, logistical regression and receiver operating characteristic curve analysis were conducted for statistical analysis. RESULTS: IDH-mutant group patients exhibited a higher calcification frequency (25.2% vs 2.4%, pâ¯=â¯0.006) and lower frequency of T1 enhancement (20.4% vs 38.1%, pâ¯=â¯0.028) comparing patients in IDH-wildtype group, while 1p/19q co-deletion group patients exhibited a higher calcification frequency (46.67% vs 2.6%, p < 0.001) and lower homogenous signal frequency in T2WI (12.0% vs 31.6%, pâ¯=â¯0.014), sharp lesion margins (14.7% vs 43.0%, pâ¯=â¯0.010), T2/fluid attenuated inversion recovery mismatch signs (22.7% vs 50.9%, pâ¯=â¯0.001), and subventricular zone involvement (64.0% vs 15.8%, pâ¯=â¯0.021) comparing patients in 1p/19q non-codeletion group. According to the results of receiver operating characteristic analysis, these features were observed to have certain diagnostic abilities, especially with regard to combination parameters, which had a high diagnostic capability, with an area under the curve of 0.848. CONCLUSION: Conventional MRI and CT features, which still represent the most convenient and widely used predictive method, might be a promising noninvasive predictor for differentiating between varied WHO grade II gliomas. Patients with calcification and T1 nonenhancement are more likely to be IDH-mutant. Moreover, patients with noncalcification, homogenous signal, sharp lesion margins, subventricular zone involvement on T2 and T2/fluid attenuated inversion recovery mismatch signs are more likely to be 1p/19q non-codeletion.
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Neoplasias Encefálicas , Glioma , Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/genética , Glioma/diagnóstico por imagen , Glioma/genética , Humanos , Isocitrato Deshidrogenasa/genética , Imagen por Resonancia Magnética , Mutación , Estudios Retrospectivos , Tomografía Computarizada por Rayos X , Organización Mundial de la SaludRESUMEN
Determining the rupture source is imperative in patient with aneurysmal subarachnoid hemorrhage (SAH). About one third of SAH cases with multiple intracranial aneurysms cannot be certain of the rupture source according to the hemorrhage pattern. This study aims to identify of the rupture source in patients with multiple intracranial aneurysms by fusing SAH data and computed tomography angiography (CTA) data. This retrospective study included 52 aneurysmal SAH patients with multiple intracranial aneurysms. In the 52 patients, 36 had definitive hemorrhage patterns on computed tomography imaging. And the other 16 patients had non-definitive hemorrhage patterns, which were bewildered for us to determine the ruptured aneurysms. Fusion of SAH data and CTA data was performed to demonstrate the spatial relationship between the SAH with each aneurysm by using the 3D Slicer software. For the patients with definitive bleed patterns, all of the suspected ruptured aneurysms were confirmed to be accurate according to the surgical records. Interestingly, the suspected rupture sources were correct in 14 of 16 patients with non-definitive hemorrhage patterns. For all 52 patients with multiple intracranial aneurysms, the ruptured aneurysms were identified in 50 cases (96.2%). In conclusion, fusion of SAH data and CTA data can precisely demonstrate the spatial relationship between the SAH with each aneurysm, which is helpful to determine the ruptured aneurysm in patients with multiple intracranial aneurysms.
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Aneurisma Roto/diagnóstico por imagen , Angiografía Cerebral/estadística & datos numéricos , Angiografía por Tomografía Computarizada/estadística & datos numéricos , Análisis de Datos , Aneurisma Intracraneal/diagnóstico por imagen , Hemorragia Subaracnoidea/diagnóstico por imagen , Adulto , Anciano , Aneurisma Roto/cirugía , Angiografía Cerebral/métodos , Angiografía por Tomografía Computarizada/métodos , Femenino , Humanos , Aneurisma Intracraneal/cirugía , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Hemorragia Subaracnoidea/cirugía , Tomografía Computarizada por Rayos X/métodos , Tomografía Computarizada por Rayos X/estadística & datos numéricosRESUMEN
Aim: In the present study, we studied the relationship between RELN and prognosis in glioma. Materials & methods: Expression profiles and methylation data of RELN were obtained from bioinformatic datasets. Correlations between RELN and clinicopathological features and overall survival were respectively assessed using chi-square test and Kaplan-Meier analysis. Results:RELN was downregulated in glioma, and its downregulation correlated well with glioma malignancy and overall survival. Meanwhile, hypermethylation of RELN was significantly correlated with low RELN expression. Additionally, gene set enrichment analysis demonstrated that low expression of RELN correlated with many key cancer pathways, possibly highlighting the importance of RELN in carcinogenesis of brain. Conclusion:RELN may serve as a potential prognostic marker and promising target molecule for new therapy of glioma.
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Neoplasias Encefálicas/genética , Moléculas de Adhesión Celular Neuronal/genética , Regulación hacia Abajo , Proteínas de la Matriz Extracelular/genética , Perfilación de la Expresión Génica/métodos , Regulación Neoplásica de la Expresión Génica , Glioma/genética , Proteínas del Tejido Nervioso/genética , Serina Endopeptidasas/genética , Adulto , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/terapia , Metilación de ADN , Femenino , Glioma/diagnóstico , Glioma/terapia , Humanos , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Pronóstico , Proteína ReelinaRESUMEN
High mobility group box 1 (HMGB1) is an abundant non-histone nuclear protein that functions as a structural protein of chromatin, regulating genome replication and recombination, mRNA transcription and DNA repair. HMGB1 has been implicated in the tumorigenesis of various cancer types, and the upregulation of HMGB1 has been demonstrated in glioma cells. However, the association between HMGB1 and the mitotic chromosomes in glioma remains uncharacterized. In the present study, the sub-cellular localization of HMGB1 in glioma tissues and cells was investigated. In addition, enhanced green fluorescent protein (EGFP)-tagging of the human HMGB1 protein and chromosome spreading were used to investigate the combination of HMGB1 with mitotic chromosomes. The results of the current study indicated that HMGB1 was localized to the nucleus and the cytoplasm, and it was determined to combine with the condensed chromosomes of proliferating cells in paraformaldehyde (PFA)-fixed glioma tissues. However, HMGB1 was also associated with interphase (but not mitotic chromosomes) when fixed with chilled methanol and 5% (v/v) acetic acid or PFA in vitro. Data from live cell imaging and chromosome spreading indicated the association of HMGB1 with mitotic chromosomes in glioma cells. The present results suggest that HMGB1 combines with mitotic chromosomes in glioma cells, and that the use of fixatives may result in the dissociation of the HMGB1-DNA interaction. Therefore, in live specimens and chromosome spreads, EGFP fusion proteins may represent an accurate indicator for the determination of the correct localization and interaction of HMGB1 in glioma cells.
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BACKGROUND: The suboccipital midline approach is common dealing with posterior fossa tumors but has a high risk of postoperative complications, such as pseudomeningocele, cerebrospinal fluid (CSF) leak, and meningitis. Neurosurgeons used various kinds of method to lower its rate. METHODS: A retrospective, single-center review of patients diagnosed with posterior fossa tumor underwent a suboccipital midline approach. Compare the rates of pseudomeningocele, CSF leak, and meningitis between 2 groups (artificial dura mater or cervical fascia autograft). We get the cervical fascia autograft from the superficial layer of deep cervical fascia just above the trapezius. RESULTS: Our retrospective review involved 123 patients matching the inclusion criteria between January 2009 and April 2019. The complication rate of pseudomeningocele, CSF leak and meningitis were 8.9%, 4.9%, and 17.9%, respectively. The presence of pseudomeningocele or CSF leak for group "artificial" was 11 of 75 (14.67%) and for group "autograft" it was 3 of 48 (6.25%). The rate of meningitis for group "artificial" (24.0%, 18 of 75) was significantly higher (P = 0.027) than the one for group "autograft" (8.33%, 4 of 48). Multivariate regression analysis suggested that the age was negatively correlated with postoperative pseudomeningocele or CSF leak (P = 0.006), with meningitis (P < 0.001). Using cervical fascia autograft decreased the rate of meningitis (P = 0.021) while showing no statistically significant clinical impact on pseudomeningocele or CSF leak. CONCLUSIONS: Applying the cervical fascia autograft to reconstruct the dura during posterior fossa surgery is a simple and effective method to reduce the rate of meningitis as compared with artificial dura mater.
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Materiales Biocompatibles/uso terapéutico , Duramadre/cirugía , Fascia/trasplante , Neoplasias Infratentoriales/cirugía , Procedimientos Neuroquirúrgicos/métodos , Procedimientos de Cirugía Plástica/métodos , Complicaciones Posoperatorias/epidemiología , Adolescente , Adulto , Anciano , Astrocitoma/cirugía , Neoplasias del Tronco Encefálico/cirugía , Neoplasias Cerebelosas/cirugía , Neoplasias del Ventrículo Cerebral/cirugía , Pérdida de Líquido Cefalorraquídeo/epidemiología , Niño , Preescolar , Ependimoma/cirugía , Femenino , Cuarto Ventrículo , Hemangioblastoma/cirugía , Hemangioma Cavernoso del Sistema Nervioso Central/cirugía , Humanos , Lactante , Masculino , Meduloblastoma/cirugía , Meningioma/cirugía , Meningitis/epidemiología , Meningocele/epidemiología , Persona de Mediana Edad , Cuello , Estudios Retrospectivos , Trasplante Autólogo , Adulto JovenRESUMEN
RATIONALE AND OBJECTIVES: Different histology and gene status of gliomas results in different natural history, treatment, and prognosis in different subgroups. Low-grade gliomas (LGGs) with isocitrate dehydrogenase (IDH) mutant and 1p/19q-codeleted are kind of gliomas with the most favorable outcome, reflecting operational strategy. Less invasive method for prediction of pathological type-even gene status-is desired. MATERIALS AND METHODS: This study investigates the potential ability of methionine-positron emission tomography (MET-PET) to determine LGGs with IDH-mutant and 1p/19q-codeleted through a retrospective review of information of 70 glioma patients. Patients underwent preoperative MET-PET, followed by operation and histopathological analysis including Immunohistochemistry and polymerase chain reaction analysis for IDH-mutant and fluorescence capillary electrophoresis analysis for 1p/19q codeletion. Texture analysis was performed for further data mining. The t-test and receiver operating characteristic curve analysis were conducted for statistical analysis. RESULTS: In the whole cohort analysis, SUVmax, SUVmean and texture features (SD and median) of oligodendroglioma, IDH-mutant and 1p/19q-codeleted patients were lower than these values of other patients. In WHO grade II subgroup analysis, no statistical difference of conventional features was observed between groups. Texture analysis displayed higher diffEntropy, diffVariance, and entropy in oligodendroglioma, IDH-mutant and 1p/19q-codeleted patients. Receiver operating characteristic analysis suggested AUCs of some conventional features and texture features ranged from 0.722 to 0.892 that are effective for diagnosis, determining LGGs with IDH-mutant and 1p/19q-codeleted in this cohort and WHO II grade glioma subgroup analysis respectively. CONCLUSION: 11C-Methionine integrated PET/MRI based texture analysis and conventional features may be a promising noninvasive predictor for differentiating the varied gliomas.
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Neoplasias Encefálicas , Glioma , Oligodendroglioma , Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/genética , Glioma/diagnóstico por imagen , Glioma/genética , Humanos , Isocitrato Deshidrogenasa/genética , Imagen por Resonancia Magnética , Metionina , Mutación , Oligodendroglioma/diagnóstico por imagen , Oligodendroglioma/genética , Tomografía de Emisión de Positrones , Estudios RetrospectivosRESUMEN
Aim: We aimed to characterize the role of HMGB1 overexpression in glioma and to evaluate its use as a biomarker. Materials & methods: We used the gene expression datasets and tissue microarray to assess the expression levels of HMGB1 among gliomas of all grades; We then assessed its correlation with the malignancy and outcome of glioma. Results: The increase in HMGB1 mRNA and protein levels was found in glioma, but there was no correlation between HMGB1 expression and glioma malignancy, and overall survival and vital status of glioma patients. Conclusion: Overexpression of HMGB1 is not associated with the malignancy and outcome in glioma. And it is not the valuable biomarker for the early diagnosis of glioma.
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Neoplasias Encefálicas/patología , Glioma/patología , Proteína HMGB1/metabolismo , Adulto , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/mortalidad , Femenino , Glioma/metabolismo , Glioma/mortalidad , Proteína HMGB1/genética , Humanos , Isocitrato Deshidrogenasa/genética , Isocitrato Deshidrogenasa/metabolismo , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Análisis Multivariante , Mutación , Clasificación del Tumor , Pronóstico , ARN Mensajero/metabolismoRESUMEN
BACKGROUND: Idiopathic hypertrophic pachymeningitis is a rare inflammatory condition with diffuse thickening of the dura mater, which may cause a compressive effect or vascular compromise. CASE DESCRIPTION: A 40-year-old Chinese man presented with persistent headache for 6 months and a sudden epileptic seizure 2 days ago. Magnetic resonance imaging demonstrated a large (71 × 34 × 27 mm) extra-axial mass at the right frontal convexity with severe edema mimicking meningioma. The lesion and peripheral dura mater showed contrast enhancement. Additionally, the skull near the lesion was eroded. Meningioma was diagnosed, and the patient underwent surgery. During the operation, we found the lesion texture was very tough, and the superior sagittal sinus was occluded. Histopathologic findings revealed a large number of infiltrated lymphocytes with fibrosis and microabscess formation; intracranial idiopathic hypertrophic pachymeningitis was diagnosed. Follow-up magnetic resonance imaging performed 3 months after surgery demonstrated the enhancement was notably alleviated. CONCLUSIONS: Idiopathic hypertrophic pachymeningitis should be part of the differential diagnosis of some cases of meningioma.
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Duramadre/patología , Neoplasias Meníngeas/diagnóstico , Meningioma/diagnóstico , Meningitis/diagnóstico , Trombosis del Seno Sagital/diagnóstico , Adulto , Diagnóstico Diferencial , Trastornos de Cefalalgia/etiología , Humanos , Hipertrofia/complicaciones , Hipertrofia/diagnóstico , Masculino , Meningitis/complicaciones , Convulsiones/etiología , Seno Sagital SuperiorRESUMEN
Inherited retinopathies typically lead to photoreceptor loss and severe visual impairments in the subjects. Intranasal administration is an efficient approach to deliver therapeutic agents to the targeted tissue. The present study is designed to deliver the erythropoietin (EPO) into the N-methyl-N-nitrosourea (MNU) induced mice, a pharmacological retinopathy model via intranasal or intravenous route. The mice were then subjected to bioavailability assay and therapeutic effects evaluation. Our results showed that the intranasal delivery of EPO is effective to alleviate the morphological disruptions in the MNU induced mice. The intranasal delivery of EPO also ameliorated the visual impairments in the MNU induced mice. Immunostaining experiment showed that both the M-cone and S-cone populations in the degenerative retinas are rescued by the intranasal delivery of EPO. In particular, the M-cone photoreceptors in dorsal-temporal (DT) quadrant and the S-cone photoreceptors in ventral-nasal (VN) quadrant were preferentially preserved by the intranasal delivery of EPO. Mechanism studies showed that the intranasal delivery of EPO could the modulate apoptosis and restrict oxidation in the degenerative retina. Compared with intravenous delivery, the intranasal delivery led to the significantly higher EPO concentration in the retina. The intranasal delivery resulted in more potent protection and had less erythropoiesis-stimulating activity than the intravenous delivery. Our results suggest that the intranasal administration is a noninvasive and efficient approach to deliver EPO into the retinas. These findings lay the groundwork for further intranasal administration of EPO in ophthalmological practice.
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Sistemas de Liberación de Medicamentos/métodos , Eritropoyetina/administración & dosificación , Células Fotorreceptoras Retinianas Conos/efectos de los fármacos , Degeneración Retiniana/tratamiento farmacológico , Administración Intranasal , Animales , Eritropoyetina/metabolismo , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Distribución Aleatoria , Retina/efectos de los fármacos , Retina/metabolismo , Retina/patología , Células Fotorreceptoras Retinianas Conos/metabolismo , Células Fotorreceptoras Retinianas Conos/patología , Degeneración Retiniana/metabolismo , Degeneración Retiniana/patologíaRESUMEN
Melatonin is a circadian hormone with potent cytoprotective effects. Retinitis pigmentosa (RP) comprises a heterogeneous group of inherent retinopathies that characterized by the photoreceptor death in bilateral eyes. The N-methyl-N-nitrosourea (MNU) administered mouse is a type of chemically induced RP model with rapid progressive rate. We intend to study the melatonin mediated effects on the MNU administered mice. Melatonin was delivered into the vitreous body of the MNU administered mice. Subsequently, the melatonin treated mice were subjected to histological analysis, optokinetic behavior tests, spectral-domain optical coherence tomography (SD-OCT), and electroretinogram (ERG) examination. Multi-electrodes array (MEA) was used to analyze the status of visual signal transmission within retinal circuits. Biochemical analysis was performed to quantify the expression levels of antioxidative enzymes, oxidative stress markers, and apoptotic factors in the retinas. The intravitreal injection of melatonin ameliorated effectively the MNU induced photoreceptor degeneration. Melatonin therapy mitigated the spontaneous firing response, and preserved the basic configurations of visual signal pathway in MNU administered mice. MEA is effective to evaluate the pharmacological effects on retina. Of note, the cone photoreceptors in degenerative retinas were rescued efficiently by melatonin therapy. Melatonin afforded these protective effects by modulating the apoptotic cascades and alleviating the oxidative stress. These findings suggest that melatonin could act as an alternative treatment for degenerative retinopathy. Melatonin might be used in combination with other therapeutic approaches to alleviate the photoreceptor loss and preserve the visual function of RP patients.
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BACKGROUND/AIMS: αB -crystallin (αBC) belongs to the family of small heat shock proteins that are necessary for maintaining oxygen homeostasis. This study was designed to explore the possible effects of αBC on N-methyl- N-nitrosourea (MNU) induced retinal degeneration and the underlying mechanisms. METHODS: The αBC was injected into the vitreous bodies of MNU administered mice. The retinal morphology and visual function of experimental animals were analyzed by electroretinography (ERG), Spectral domain optical coherence tomography (SD-OCT), fundus photographs, optokinetic testing and immunohistochemistry assay. RESULTS: Optokinetic behavioural tests and ERG examination suggested that the visual impairments of the MNU administered mice were ameliorated effectively by αBC treatment. OCT analysis showed that the major retinal architecture of the MNU administered mice was efficiently rescued by αBC treatment. Fundus examination suggested that the lesion size of the MNU administered mice was decreased by αBC treatment. MNU induced photoreceptor loss was also mitigated by αBC treatment as shown by hematoxylin and eosin staining. In particular, the immunostaining study suggested that M-cone photoreceptors, rather than the S-cone photoreceptors, were preferentially rescued, indicating that the photoreceptor populations have different sensitivities to αBC. The mechanism study suggested that the anti-apoptotic, anti-oxidative and neurotrophic function of αBC collectively contributed to these therapeutic effects. CONCLUSION: Intravitreal delivery of αBC could alleviate MNU induced photoreceptor degeneration and visual impairment. Further refinement of the αBC induced protection would afford a novel therapeutic strategy for retinitis pigmentosa.
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Cristalinas/metabolismo , Metilnitrosourea/toxicidad , Degeneración Retiniana/patología , Animales , Caspasa 3/metabolismo , Cristalinas/genética , Modelos Animales de Enfermedad , Electrorretinografía , Inyecciones Intravítreas , Malondialdehído/metabolismo , Ratones , Ratones Endogámicos C57BL , Células Fotorreceptoras/metabolismo , Retina/diagnóstico por imagen , Retina/metabolismo , Degeneración Retiniana/inducido químicamente , Superóxido Dismutasa/metabolismo , Tomografía de Coherencia Óptica , Proteína X Asociada a bcl-2/metabolismoRESUMEN
The inflammatory response following spinal cord injury (SCI) involves the activation of resident microglia and the infiltration of macrophages. Macrophages and microglia can be polarized into the classically activated proinflammatory M1 phenotype or the alternatively activated anti-inflammatory M2 phenotype. Programmed cell death 1 (PD-1) is a critical immune inhibitory receptor involved in innate and adaptive immune responses. However, whether PD-1 is involved in the modulation of macrophage/microglial polarization is unknown. In this study, the mRNA levels of pd1 gradually increased after SCI, and PD-1 protein was found in macrophages/microglia in injured spinal cord sections. PD-1 knockout (KO) mice showed poor locomotor recovery after spinal cord crushing compared with wild-type mice. M1-type macrophages/microglia accumulated in greater numbers in the injured spinal cord of PD-1-KO mice. Under polarized stimulation, induced expression of PD-1 occurred in cultured macrophages and microglia. PD-1 suppressed M1 polarization by reducing the phosphorylation of signal transducer and activator of transcription 1 (STAT1) and promoted M2 polarization by increasing STAT6 phosphorylation. In PD-1-KO mice, the M1 response was enhanced via the activation of STAT1 and nuclear factor-kappa B. Furthermore, PD-1 played various roles in phagocytosis in macrophages and microglia. Therefore, our results suggest that PD-1 signaling plays an important role in the regulation of macrophage/microglial polarization. Thus, deregulated PD-1 signaling may induce the polarization of macrophages/microglia toward the M1 phenotype. Overall, our results provide new insights into the modulatory mechanisms of macrophage/microglial polarization, thereby possibly facilitating the development of new therapies for SCI via the regulation of macrophage/microglial polarization through PD-1 signaling.
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Macrófagos/citología , Macrófagos/metabolismo , Microglía/citología , Microglía/metabolismo , Receptor de Muerte Celular Programada 1/metabolismo , Traumatismos de la Médula Espinal/metabolismo , Animales , Polaridad Celular , Inflamación/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , FN-kappa B/metabolismo , Fenotipo , Cultivo Primario de Células , Receptor de Muerte Celular Programada 1/genética , ARN Mensajero/metabolismo , Factor de Transcripción STAT1/metabolismo , Factor de Transcripción STAT6/metabolismoRESUMEN
BACKGROUND: Hemorrhage is a direct consequence of traumatic injury to the central nervous system and may cause innate immune reactions including cerebral Toll-like receptor (TLR) 4 upregulation which usually leads to poor outcome in the traumatic brain injury. In spinal cord injury (SCI), however, how hemorrhage induces innate immune reaction in spinal parenchyma remains unknown. The present study aimed to see whether blood component and/or other factor(s) induce TLR4 and microglia/macrophages involved innate immune reactions in the rat spinal cord after traumatic injury. METHODS: Using the compressive SCI model of the rat, hemorrhage in the spinal cord was identified by hematoxylin-eosin staining. Microglia/macrophage activation, TLR4 expression, and cell apoptosis were investigated by immunohistochemistry. Nuclear factor (NF)-κB p50 level of the two segments of the cord was detected by western blotting assay. With carbon powder injection, blood origination of the hematoma was explored. The blood-spinal cord barrier (BSCB) states of the lesion site and the hematoma were compared with immunohistochemistry and tannic acid-ferric chloride staining. RESULTS: Histological observation found blood accumulated in the center of compression lesion site (epicenter) and in the hematoma approximately 1.5 cm away from the epicenter. TLR4 expression, microglia//macrophage activation, and subsequent apoptosis in the area of far-away hematoma were late and weak in comparison to that in epicenter. In addition, TLR4 positive microglia/macrophages appeared to be phagocytotic in the far-away hematoma more obviously than that in the epicenter. Injected carbon powder indicated that accumulated blood of the far-away hematoma originated from the bleeding of the lesion epicenter, and the BSCB around the hematoma was not compromised in the early phase. Accordingly, at 3 days post injury, NF-κB p50 was upregulated based on the similar levels of blood component hemoglobin, and cell apoptosis was obvious in the epicenter but not in the far-away hematoma. CONCLUSION: These data suggest that besides blood component, BSCB compromise and the extent of tissue injury contribute more to TLR4 and microglia/macrophage responses to the spinal cord hemorrhage. Therefore, the innate immune environment is a necessary consideration for the SCI therapy targeting TLR4 and microglia/macrophages.
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Activación de Macrófagos/inmunología , Macrófagos/metabolismo , Microglía/inmunología , Compresión de la Médula Espinal/inmunología , Compresión de la Médula Espinal/patología , Receptor Toll-Like 4/biosíntesis , Animales , Western Blotting , Modelos Animales de Enfermedad , Hemorragia/inmunología , Hemorragia/metabolismo , Hemorragia/patología , Macrófagos/inmunología , Masculino , Microglía/metabolismo , Ratas , Ratas Sprague-Dawley , Compresión de la Médula Espinal/metabolismoRESUMEN
To determine the role of toll-like receptors (TLRs) myeloid differentiation factor 88 (MyD88) dependent pathway in the spinal cord secondary injury, compression injury was made at T8 segment of the spinal cord in adult male Sprague-Dawley rats. Shown by RT-PCR, TLR4 mRNA in the spinal cord was quickly elevated after compression injury. Intramedullary injection of MyD88 inhibitory peptide (MIP) resulted in significant improvement in locomotor function recovery at various time points after surgery. Meanwhile, injury area, p38 phosphorylation, and proinflammation cytokines in the injured spinal cord were significantly reduced in MIP-treated animals, compared with control peptide (CP) group. These data suggest that TLRs MyD88-dependent pathway may play an important role in the development of secondary spinal cord injury, and inhibition of this pathway at early time after primary injury could effectively protect cells from inflammation and apoptosis and therefore improve the functional recovery.
RESUMEN
Inosine is a purine nucleoside and is considered protective to neural cells including neurons and astrocytes against hypoxic injury. However, whether oligodendrocytes (OLs) could also be protected from hypoxia by inosine is not known. Here we investigated the effects of inosine on primarily cultured rat OLs injured by rotenone-mediated chemical hypoxia, and the mechanisms of the effects using ATP assay, MTT assay, PI-Hoechst staining, TUNEL, and immunocytochemistry. Results showed that rotenone exposure for 24 h caused cell death and impaired viability in both immature and mature OLs, while pretreatment of 10 mM inosine 30 min before rotenone administration significantly reduced cell death and improved the viability of OLs. The same concentration of inosine given 120 min after rotenone exposure also improved viability of injured mature OLs. Immunocytochemistry for nitrotyrosine and cellular ATP content examination indicated that inosine may protect OLs by providing ATP and scavenging peroxynitrite for cells. In addition, immature OLs were more susceptible to hypoxia than mature OLs; and at the similar degree of injury, inosine protected immature and mature OLs differently. Quantitative real-time PCR revealed that expression of adenosine receptors was different between these two stages of OLs. These data suggest that inosine protect OLs from hypoxic injury as an antioxidant and ATP provider, and the protective effects of inosine on OLs vary with cell differentiation, possibly due to the adenosine receptors expression profile. As OLs form myelin in the central nervous system, inosine could be used as a promising drug to treat demyelination-involved disorders.
Asunto(s)
Hipoxia/inducido químicamente , Inosina/farmacología , Insecticidas/farmacología , Oligodendroglía/efectos de los fármacos , Rotenona/farmacología , Adenosina Trifosfato/metabolismo , Animales , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Etiquetado Corte-Fin in Situ , Oligodendroglía/citología , RatasRESUMEN
Shu-Xue-Tong (SXT) is a traditional Chinese drug widely used to ameliorate stagnation of blood flow, such as brain or myocardial infarction. Whether SXT may have therapeutic value for spinal cord injury (SCI), during which ischemia plays an important role in its pathology, remains to be elucidated. We hypothesized that SXT may promote SCI healing by improving spinal cord blood flow (SCBF), and a study was thus designed to explore this possibility. Twenty-five male Sprague-Dawley rats were used. SCI was induced by compression, and SXT was administrated 24 h postinjury for 14 successive days. The effects of SXT were assessed by means of laser-Doppler flowmetry, motor functional analysis (open-field walking and footprint analysis), and histological analysis (hematoxylin-eosin and thionin staining and NeuN immunohistochemistry). SXT significantly promoted SCBF of the contused spinal cord and enhanced the recovery of motor function. Histological analysis indicated that the lesion size was reduced, the pathological changes were ameliorated, and more neurons were preserved. Based on these results we conclude that SXT can effectively improve SCI.