RESUMO
BACKGROUND AND PURPOSE: Liver fibrosis, a common yet often subclinical manifestation of chronic liver disease, may have an unrecognized role in cognitive impairment. We evaluated the association between a validated liver fibrosis index and cognitive measures among older adults. METHODS: We examined the association between liver fibrosis and cognitive performance among participants aged 60 years and older in the US National Health and Nutrition Examination Survey. Liver fibrosis was measured with the validated Fibrosis-4 (FIB-4) liver fibrosis score. The outcomes were performance on four standardized cognitive tests of immediate and delayed verbal learning, verbal fluency, and attention/concentration. We used linear regression to evaluate the association between FIB-4 score and performance on cognitive tests while adjusting for potential confounders. In sensitivity analyses, we examined this association in participants without known liver disease. RESULTS: Among 3217 adult participants, the mean age was 69 years, and 54% were women. Standard liver chemistries were largely in the normal range. However, 5.0% [95% confidence interval (CI) 4.0-6.0] had liver fibrosis based on a validated cut-off. In adjusted linear regression models, higher liver fibrosis scores were associated with worse immediate recall (ß -0.39; 95% CI -0.58, -0.21), language fluency (ß -0.46; 95% CI -0.72, -0.21), and attention/concentration (ß -1.34; 95% CI -2.25, -0.43), but not delayed recall (ß -0.10; 95% CI -0.20, 0.01). Results were similar when limiting the study population to participants without known clinical liver disease. CONCLUSION: Liver fibrosis, including subclinical liver fibrosis, may be an independent risk factor for cognitive impairment among older adults.
Assuntos
Cognição , Disfunção Cognitiva , Cirrose Hepática , Idoso , Disfunção Cognitiva/epidemiologia , Feminino , Humanos , Cirrose Hepática/complicações , Cirrose Hepática/epidemiologia , Masculino , Pessoa de Meia-Idade , Testes Neuropsicológicos , Inquéritos NutricionaisRESUMO
BACKGROUND AND PURPOSE: Calcification of the intracranial vasculature is an independent risk factor for stroke. The relationship between luminal stenosis and calcium burden in the intracranial circulation is incompletely understood. We evaluated the relationship between atherosclerotic calcification and luminal stenosis in the intracranial ICAs. MATERIALS AND METHODS: Using a prospective stroke registry, we identified patients who had both NCCT and CTA or MRA examinations as part of a diagnostic evaluation for ischemic stroke. We used NCCTs to qualitatively (modified Woodcock Visual Score) and quantitatively (Agatston-Janowitz Calcium Score) measure ICA calcium burden and used angiography to measure arterial stenosis. We calculated correlation coefficients between the degree of narrowing and calcium burden measures. RESULTS: In 470 unique carotid arteries (235 patients), 372 (79.1%) had atherosclerotic calcification detectable on CT compared with 160 (34%) with measurable arterial stenosis on CTA or MRA (P < .001). We found a weak linear correlation between qualitative (R = 0.48) and quantitative (R = 0.42) measures of calcium burden and the degree of luminal stenosis (P < .001 for both). Of 310 ICAs with 0% luminal stenosis, 216 (69.7%) had measurable calcium scores. CONCLUSIONS: There is a weak correlation between intracranial atherosclerotic calcium scores and luminal narrowing, which may be explained by the greater sensitivity of CT than angiography in detecting the presence of measurable atherosclerotic disease. Future studies are warranted to evaluate the relationship between stenosis and calcium burden in predicting stroke risk.
Assuntos
Calcinose/diagnóstico por imagem , Cálcio/metabolismo , Arteriosclerose Intracraniana/diagnóstico por imagem , Idoso , Idoso de 80 Anos ou mais , Isquemia Encefálica/diagnóstico por imagem , Calcinose/metabolismo , Artéria Carótida Interna/diagnóstico por imagem , Estenose das Carótidas/diagnóstico por imagem , Angiografia Cerebral , Feminino , Humanos , Arteriosclerose Intracraniana/metabolismo , Angiografia por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Reprodutibilidade dos Testes , Medição de Risco , Acidente Vascular Cerebral/diagnóstico por imagem , Tomografia Computadorizada por Raios XRESUMO
INTRODUCTION: Breast cancer is the most common cancer in women and clearly increases the risk of venous thromboembolism. However, its association with arterial thromboembolism is less well defined. AIM: To determine the short-term cumulative incidence and relative hazard of arterial thromboembolism in elderly patients with incident breast cancer. MATERIALS AND METHODS: Using the Surveillance Epidemiology and End Results-Medicare linked database, which includes approximately 28% of all patients diagnosed with cancer in the United States, we identified patients with a new primary diagnosis of breast cancer from 2002 through 2011. These patients were individually matched by age, sex, race, registry, and medical comorbidities to a group of Medicare enrollees without cancer, and each pair was followed through 2012. Validated diagnosis codes were used to identify a primary composite outcome of arterial thromboembolism defined as any ischemic stroke or myocardial infarction. Secondary outcomes included ischemic stroke alone and myocardial infarction alone. Cumulative incidence rates were calculated using competing risk survival statistics. The Gray test was used to compare rates between groups. The proportional hazard assumption was violated for the entirety of patient follow-up; therefore, Cox proportional hazard analysis was performed at discrete time points when the assumption was generally met. RESULTS: We identified 96,666 pairs of breast cancer patients and matched controls. Median age was 75 years and few cancers were advanced at diagnosis (12% stages 3/4). The 3-month cumulative incidence of arterial thromboembolism was 2.1% (95% confidence interval [CI] 2.0-2.2%) in cancer patients compared to 1.4% (95% CI 1.3-1.5%) in controls (hazard ratio [HR] 1.5, 95% CI 1.4-1.6, p<0.01). The short-term risk of each secondary outcome was heightened in the breast cancer group, although the relative hazard for myocardial infarction was higher than for ischemic stroke. The 3-month cumulative incidence of ischemic stroke was 1.3% (95% CI 1.2-1.4%) in cancer patients compared to 1.0% (95% CI 0.9-1.1%) in controls (HR 1.3, 95% CI 1.2-1.4, p<0.01), and the 3-month cumulative incidence of myocardial infarction was 0.9% (95% CI 0.8-0.9%) in cancer patients compared to 0.4% (0.4-0.5%) in controls (HR 2.0, 95% CI 1.8-2.3, p<0.01). Excess risks attenuated over time and were no longer present beyond 1 year. CONCLUSIONS: Patients with incident breast cancer face an increased short-term risk of ischemic stroke and myocardial infarction.
RESUMO
There are profound, yet incompletely understood, sex differences in the neurogenic regulation of blood pressure. Both corticotropin signaling and glutamate receptor plasticity, which differ between males and females, are known to play important roles in the neural regulation of blood pressure. However, the relationship between hypertension and glutamate plasticity in corticotropin-releasing factor (CRF)-receptive neurons in brain cardiovascular regulatory areas, including the rostral ventrolateral medulla (RVLM) and paraventricular nucleus of the hypothalamus (PVN), is not understood. In the present study, we used dual-label immuno-electron microscopy to analyze sex differences in slow-pressor angiotensin II (AngII) hypertension with respect to the subcellular distribution of the obligatory NMDA glutamate receptor subunit 1 (GluN1) subunit of the N-methyl-D-aspartate receptor (NMDAR) in the RVLM and PVN. Studies were conducted in mice expressing the enhanced green fluorescence protein (EGFP) under the control of the CRF type 1 receptor (CRF1) promoter (i.e., CRF1-EGFP reporter mice). By light microscopy, GluN1-immunoreactivity (ir) was found in CRF1-EGFP neurons of the RVLM and PVN. Moreover, in both regions tyrosine hydroxylase (TH) was found in CRF1-EGFP neurons. In response to AngII, male mice showed an elevation in blood pressure that was associated with an increase in the proportion of GluN1 on presumably functional areas of the plasma membrane (PM) in CRF1-EGFP dendritic profiles in the RVLM. In female mice, AngII was neither associated with an increase in blood pressure nor an increase in PM GluN1 in the RVLM. Unlike the RVLM, AngII-mediated hypertension had no effect on GluN1 localization in CRF1-EGFP dendrites in the PVN of either male or female mice. These studies provide an anatomical mechanism for sex-differences in the convergent modulation of RVLM catecholaminergic neurons by CRF and glutamate. Moreover, these results suggest that sexual dimorphism in AngII-induced hypertension is reflected by NMDA receptor trafficking in presumptive sympathoexcitatory neurons in the RVLM.
Assuntos
Hipertensão/patologia , Bulbo/citologia , Proteínas do Tecido Nervoso/metabolismo , Plasticidade Neuronal/genética , Neurônios/metabolismo , Receptores de Hormônio Liberador da Corticotropina/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Caracteres Sexuais , Angiotensina II/toxicidade , Animais , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/genética , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Hipertensão/induzido quimicamente , Hipertensão/genética , Masculino , Bulbo/efeitos dos fármacos , Camundongos , Camundongos Transgênicos , Microscopia Imunoeletrônica , Proteínas do Tecido Nervoso/genética , Plasticidade Neuronal/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurônios/ultraestrutura , RNA Mensageiro/metabolismo , Receptores de Hormônio Liberador da Corticotropina/genética , Receptores de N-Metil-D-Aspartato/genética , Estilbamidinas/metabolismo , Frações Subcelulares/metabolismo , Frações Subcelulares/ultraestrutura , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Alzheimer's dementia is a devastating and incurable disease afflicting over 35 million people worldwide. Amyloid-ß (Aß), a key pathogenic factor in this disease, has potent cerebrovascular effects that contribute to brain dysfunction underlying dementia by limiting the delivery of oxygen and glucose to the working brain. However, the downstream pathways responsible for the vascular alterations remain unclear. Here we report that the cerebrovascular dysfunction induced by Aß is mediated by DNA damage caused by vascular oxidative-nitrosative stress in cerebral endothelial cells, which, in turn, activates the DNA repair enzyme poly(ADP)-ribose polymerase. The resulting increase in ADP ribose opens transient receptor potential melastatin-2 (TRPM2) channels in endothelial cells leading to intracellular Ca(2+) overload and endothelial dysfunction. The findings provide evidence for a previously unrecognized mechanism by which Aß impairs neurovascular regulation and suggest that TRPM2 channels are a potential therapeutic target to counteract cerebrovascular dysfunction in Alzheimer's dementia and related pathologies.
Assuntos
Sistema Nervoso/irrigação sanguínea , Sistema Nervoso/fisiopatologia , Canais de Cátion TRPM/metabolismo , Peptídeos beta-Amiloides , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/patologia , Encéfalo/fisiopatologia , Cálcio/metabolismo , Circulação Cerebrovascular/efeitos dos fármacos , Dano ao DNA , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Glicosídeo Hidrolases/antagonistas & inibidores , Glicosídeo Hidrolases/metabolismo , Ativação do Canal Iônico/efeitos dos fármacos , Camundongos Transgênicos , Microvasos/metabolismo , Microvasos/patologia , Sistema Nervoso/metabolismo , Nitrosação/efeitos dos fármacos , Ácido Peroxinitroso/metabolismo , Inibidores de Poli(ADP-Ribose) Polimerases , Poli(ADP-Ribose) Polimerases/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Sistema Vasomotor/metabolismo , Sistema Vasomotor/fisiopatologiaRESUMO
In the central nervous system, angiotensin II (AngII) binds to angiotensin type 1 receptors (AT(1)Rs) to affect autonomic and endocrine functions as well as learning and memory. However, understanding the function of cells containing AT(1)Rs has been restricted by limited availability of specific antisera, difficulties discriminating AT(1)R-immunoreactive cells in many brain regions and, the identification of AT(1)R-containing neurons for physiological and molecular studies. Here, we demonstrate that an Agtr1a bacterial artificial chromosome (BAC) transgenic mouse line that expresses type A AT(1)Rs (AT1aRs) identified by enhanced green fluorescent protein (EGFP) overcomes these shortcomings. Throughout the brain, AT1aR-EGFP was detected in the nuclei and cytoplasm of cells, most of which were neurons. EGFP often extended into dendritic processes and could be identified either natively or with immunolabeling of GFP. The distribution of AT1aR-EGFP cells in brain closely corresponded to that reported for AngII binding and AT1aR protein and mRNA. In particular, AT1aR-EGFP cells were in autonomic regions (e.g., hypothalamic paraventricular nucleus, central nucleus of the amygdala, parabrachial nucleus, nuclei of the solitary tract and rostral ventrolateral medulla) and in regions involved in electrolyte and fluid balance (i.e., subfornical organ) and learning and memory (i.e., cerebral cortex and hippocampus). Additionally, dual label electron microscopic studies in select brain areas demonstrate that cells containing AT1aR-EGFP colocalize with AT(1)R-immunoreactivity. Assessment of AngII-induced free radical production in isolated EGFP cells demonstrated feasibility of studies investigating AT1aR signaling ex vivo. These findings support the utility of Agtr1a BAC transgenic reporter mice for future studies understanding the role of AT(1)R-containing cells in brain function.
Assuntos
Química Encefálica/genética , Encéfalo/citologia , Cromossomos Artificiais Bacterianos/genética , Receptor Tipo 1 de Angiotensina/metabolismo , Animais , Arginina Vasopressina/imunologia , Arginina Vasopressina/metabolismo , Sistema Nervoso Autônomo/citologia , Sistema Nervoso Autônomo/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/imunologia , Humanos , Processamento de Imagem Assistida por Computador , Técnicas Imunoenzimáticas , Imuno-Histoquímica , Camundongos , Camundongos Transgênicos , Microscopia Eletrônica , Microscopia Imunoeletrônica , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Espécies Reativas de Oxigênio/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Equilíbrio Hidroeletrolítico/genética , Equilíbrio Hidroeletrolítico/fisiologiaRESUMO
The rostral ventrolateral medulla (RVLM), a region critical for the tonic and reflex control of arterial pressure, contains a group of adrenergic (C1) neurons that project to the spinal cord and directly modulate pre-ganglionic sympathetic neurons. Epidemiological data suggest that there are gender differences in the regulation of blood pressure. One factor that could be involved is angiotensin II signaling and the associated production of reactive oxygen species (ROS) by NADPH oxidase, which is emerging as an important molecular substrate for central autonomic regulation and dysregulation. In this study dual electron microscopic immunolabeling was used to examine the subcellular distribution of the angiotensin type 1 (AT(1)) receptor and two NADPH oxidase subunits (p47 and p22) in C1 dendritic processes, in tissue from male, proestrus (high estrogen) and diestrus (low estrogen) female rats. Female dendrites displayed significantly more AT(1) labeling and significantly less p47 labeling than males. While elevations in AT(1) labeling primarily resulted from higher levels of receptor on the plasma membrane, p47 labeling was reduced both on the plasma membrane and in the cytoplasm. Across the estrous cycle, proestrus females displayed significantly higher levels of AT(1) labeling than diestrus females, which resulted exclusively from plasma membrane density differences. In contrast, p47 labeling did not change across the estrous cycle, indicating that ROS production might reflect AT(1) receptor membrane density. No significant differences in p22 labeling were observed. These findings demonstrate that both sex and hormonal levels can selectively affect the expression and subcellular distribution of components of the angiotensin II signaling pathway within C1 RVLM neurons. Such effects could reflect differences in the capacity for ROS production, potentially influencing short term excitability and long term gene expression in a cell group which is critically involved in blood pressure regulation, potentially contributing to gender differences in the risk of cardiovascular disease.
Assuntos
Dendritos/metabolismo , Bulbo/metabolismo , NADPH Oxidases/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Formação Reticular/metabolismo , Caracteres Sexuais , Angiotensina II/metabolismo , Animais , Pressão Sanguínea/fisiologia , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Citoplasma/metabolismo , Citoplasma/ultraestrutura , Dendritos/ultraestrutura , Ciclo Estral/fisiologia , Feminino , Hormônios Esteroides Gonadais/metabolismo , Masculino , Bulbo/ultraestrutura , Microscopia Imunoeletrônica , NADPH Oxidases/química , Estresse Oxidativo/fisiologia , Subunidades Proteicas/metabolismo , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Reprodução/fisiologia , Formação Reticular/ultraestrutura , Regulação para Cima/fisiologiaRESUMO
The hypothalamic paraventricular nucleus (PVN) and angiotensin II (AngII) play critical roles in cardiovascular and neurohumoral regulation ascribed in part to vasopressin (VP) release. The AngII actions in the PVN are mediated largely through angiotensin II type 1 (AT1) receptors. However, there is indirect evidence that the functionally elusive central angiotensin II type 2 (AT2) receptors are also mediators of AngII signaling in the PVN. We used electron microscopic dual immunolabeling of antisera recognizing the AT2 receptor and VP to test the hypothesis that mouse PVN neurons expressing VP are among the cellular sites where this receptor has a subcellular distribution conducive to local activation. Immunoreactivity for the AT2 receptor was detected in somatodendritic profiles, of which approximately 60% of the somata and approximately 28% of the dendrites also contained VP. In comparison with somata and dendrites, axons, axon terminals, and glia less frequently contained the AT2 receptor. Somatic labeling for the AT2 receptor was often seen in the cytoplasm near the Golgi lamellae and other endomembrane structures implicated in receptor trafficking. AT2 receptor immunoreactivity in dendrites was commonly localized to cytoplasmic endomembranes, but was occasionally observed on extra- or peri-synaptic portions of the plasma membrane apposed by astrocytic processes or by unlabeled axon terminals. The labeled dendritic plasmalemmal segments containing AT2 receptors received asymmetric excitatory-type or more rarely symmetric inhibitory-type contacts from unlabeled axon terminals containing dense core vesicles, many of which are known to store neuropeptides. These results provide the first ultrastructural evidence that AT2 receptors in PVN neurons expressing VP and other neuromodulators are strategically positioned for surface activation by AngII and/or intracellular trafficking.
Assuntos
Angiotensinas/metabolismo , Neurônios/metabolismo , Núcleo Hipotalâmico Paraventricular/metabolismo , Receptor Tipo 2 de Angiotensina/metabolismo , Vasopressinas/metabolismo , Animais , Astrócitos/metabolismo , Astrócitos/ultraestrutura , Compartimento Celular/fisiologia , Forma Celular/fisiologia , Citoplasma/metabolismo , Citoplasma/ultraestrutura , Dendritos/metabolismo , Dendritos/ultraestrutura , Células HeLa , Humanos , Membranas Intracelulares/metabolismo , Membranas Intracelulares/ultraestrutura , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Imunoeletrônica , Neurônios/ultraestrutura , Núcleo Hipotalâmico Paraventricular/citologia , Terminações Pré-Sinápticas/metabolismo , Terminações Pré-Sinápticas/ultraestrutura , Transporte Proteico/fisiologia , Transdução de Sinais/fisiologia , Membranas Sinápticas/metabolismo , Membranas Sinápticas/ultraestrutura , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologiaRESUMO
Women are less susceptible to the cerebrovascular complications of hypertension, such as a stroke and vascular dementia. The mechanism of such protection may be related to a reduced vulnerability of women to the cerebrovascular actions of hypertension. To test this hypothesis, we used a model of hypertension based on infusion of angiotensin II (ANG II), an octapeptide that plays a key role in hypertension and produces cerebrovascular dysregulation. Cerebral blood flow (CBF) was monitored by laser-Doppler flowmetry in anesthetized (urethane-chloralose) C57BL/6J male and female mice equipped with a cranial window. ANG II administration (0.25 mug.kg(-1).min(-1) iv x 30-45 min) elevated arterial pressure equally in both sexes but attenuated the CBF increase induced by whisker stimulation or by the endothelium-dependent vasodilator acetylcholine (ACh) in male but not in female mice. The administration of ANG II for 7 days (2.74 mg.kg(-1).day(-1)), using osmotic minipumps, also attenuated these cerebrovascular responses in male, but not female, mice. The reduced susceptibility to the effect of ANG II in female mice was abolished by ovariectomy and reinstated by estrogen administration to ovariectomized mice. Administration of estrogen to male mice abolished the ANG II-induced attenuation of CBF responses. We conclude that female mice are less susceptible to the cerebrovascular dysregulation induced by ANG II, an effect related to estrogen. Such protection from the deleterious cerebrovascular effects of hypertension may play a role in the reduced vulnerability to the cerebrovascular complications of hypertension observed in women.
Assuntos
Angiotensina II/metabolismo , Circulação Cerebrovascular , Estradiol/metabolismo , Terapia de Reposição de Estrogênios , Hipertensão/complicações , Neocórtex/irrigação sanguínea , Acidente Vascular Cerebral/etiologia , Acetilcolina/farmacologia , Adenosina/farmacologia , Angiotensina II/administração & dosagem , Animais , Velocidade do Fluxo Sanguíneo , Pressão Sanguínea , Circulação Cerebrovascular/efeitos dos fármacos , Modelos Animais de Doenças , Suscetibilidade a Doenças , Estradiol/administração & dosagem , Feminino , Hipertensão/induzido quimicamente , Hipertensão/metabolismo , Hipertensão/fisiopatologia , Bombas de Infusão Implantáveis , Fluxometria por Laser-Doppler , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neocórtex/efeitos dos fármacos , Neocórtex/metabolismo , Neocórtex/fisiopatologia , Ovariectomia , Reflexo , Fatores Sexuais , Acidente Vascular Cerebral/metabolismo , Acidente Vascular Cerebral/fisiopatologia , Fatores de Tempo , Vasodilatadores/farmacologia , Vibrissas/inervaçãoRESUMO
Superoxide produced by the enzyme nicotinamide adenine dinucleotide phosphate (NADPH) oxidase mediates crucial intracellular signaling cascades in the medial nucleus of the solitary tract (mNTS), a brain region populated by catecholaminergic neurons, as well as astroglia that play an important role in autonomic function. The mechanisms mediating NADPH oxidase (phagocyte oxidase) activity in the neural regulation of cardiovascular processes are incompletely understood, however the subcellular localization of superoxide produced by the enzyme is likely to be an important regulatory factor. We used immunogold electron microscopy to determine the phenotypic and subcellular localization of the NADPH oxidase subunits p47(phox), gp91(phox,) and p22(phox) in the mNTS in rats. The mNTS contains a large population of neurons that synthesize catecholamines. Significantly, catecholaminergic signaling can be modulated by redox reactions. Therefore, the relationship of NADPH oxidase subunit labeled neurons or glia with respect to catecholaminergic neurons was also determined by dual labeling for the superoxide producing enzyme and tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis. In the mNTS, NADPH oxidase subunits were present primarily in somatodendritic processes and astrocytes, some of which also contained TH, or were contacted by TH-labeled axons, respectively. Immunogold quantification of NADPH oxidase subunit localization showed that p47(phox) and gp91(phox) were present on the surface membrane, as well as vesicular organelles characteristic of calcium storing smooth endoplasmic reticula in dendritic and astroglial processes. These results indicate that NADPH oxidase assembly and consequent superoxide formation are likely to occur near the plasmalemma, as well as on vesicular organelles associated with intracellular calcium storage within mNTS neurons and glia. Thus, NADPH oxidase-derived superoxide may participate in intracellular signaling pathways linked to calcium regulation in diverse mNTS cell types. Moreover, NADPH oxidase-derived superoxide in neurons and glia may directly or indirectly modulate catecholaminergic neuron activity in the mNTS.
Assuntos
Astrócitos/metabolismo , NADPH Oxidases/metabolismo , Neurônios/metabolismo , Núcleo Solitário/citologia , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Astrócitos/ultraestrutura , RNA Helicases DEAD-box , Imuno-Histoquímica/métodos , Espaço Intracelular/metabolismo , Espaço Intracelular/ultraestrutura , Masculino , Microscopia Eletrônica de Transmissão/métodos , Neurônios/ultraestrutura , Proteínas Nucleares/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
The angiotensin II AT-1A receptor (AT-1A) is the major mediator of the hypertensive actions of angiotensin II (ANG II) in the medial nucleus of the solitary tract (mNTS). The localization of the AT-1A receptor at surface or intracellular sites is an important determinant of its signaling properties, including intercellular or intracrine communication. However, the spatial localization of this protein, particularly within small distal or intermediate size dendrites of mNTS neurons, is unknown. Within the mNTS, ANG II and catecholamines interact in the regulation of autonomic function; however, it is unknown if AT-1A receptors are present at functional sites in catecholamine containing dendrites, or are contacted by catecholamine containing axon terminals. We compared surface and intracellular distributions of the AT-1A receptor in dendritic processes from the mNTS using immunogold electron microscopy in conjunction with immunoperoxidase labeling for tyrosine hydroxylase (TH) and morphometric analysis. Collapsed across all AT-1A-labeled dendritic profiles, immunogold labeling was more frequent in intracellular sites as compared with the plasma membrane. Small (<0.6 microm) dendritic profiles contained a higher ratio of particles associated with the surface membrane when compared with larger profiles. Approximately 27% of all AT-1A receptor-labeled dendritic profiles also contained labeling for TH. Approximately 12% of dendritic profiles single labeled for the AT-1A receptor were contacted by TH containing axons or axon terminals. The present results provide the first quantitative demonstration of select plasmalemmal and intracellular localizations of AT-1A receptors in dendritic processes of mNTS neurons, including those containing TH, or contacted by catecholaminergic axon terminals. These results suggest that AT-1A receptors are positioned for modulation of catecholamine signaling in the mNTS.
Assuntos
Catecolaminas/metabolismo , Neurônios/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Núcleo Solitário/metabolismo , Animais , Dendritos/metabolismo , Dendritos/ultraestrutura , Técnicas Imunoenzimáticas , Imuno-Histoquímica , Masculino , Proteínas de Membrana/metabolismo , Neurônios/ultraestrutura , Terminações Pré-Sinápticas/enzimologia , Ratos , Ratos Sprague-Dawley , Núcleo Solitário/citologia , Núcleo Solitário/ultraestrutura , Frações Subcelulares/metabolismo , Frações Subcelulares/ultraestrutura , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Angiotensin II (Ang II) type 1 (AT1) receptors are prevalent in the sensory vagal complex including the nucleus tractus solitarii (NTS) and area postrema, each of which has been implicated in the central cardiovascular effects produced by Ang II. In rodents, these actions prominently involve the AT1A receptor. Thus, we examined the electron microscopic dual immunolabeling of antisera recognizing the AT1A receptor and Ang II to determine interactive sites in the sensory vagal complex of rat brain. In both the area postrema and adjacent dorsomedial NTS, many somatodendritic profiles were dually labeled for the AT1A receptor and Ang II. In these profiles, AT1A receptor-immunoreactivity was often seen in the cytoplasm beneath labeled portions of the plasma membrane and in endosome-like granules as well as Golgi lamellae and outer nuclear membranes. In addition, AT1A receptor labeling was detected on the plasma membrane and in association with cytoplasmic membranes in many small axons and axon terminals. These terminals were morphologically heterogeneous containing multiple types of vesicles and forming either inhibitory- or excitatory-type synapses. In the area postrema, AT1A receptor labeling also was detected in many non-neuronal cells including glia, capillary endothelial cells and perivascular fibroblasts that were less prevalent in the NTS. We conclude that in the rat sensory vagal complex, AT1A receptors are strategically positioned for involvement in modulation of the postsynaptic excitability and intracrine hormone-like effects of Ang II. In addition, these receptors have distributions consistent with diverse roles in regulation of transmitter release, regional blood flow and/or vascular permeability.
Assuntos
Angiotensina II/análise , Área Postrema/química , Gânglios Sensitivos/química , Receptor Tipo 1 de Angiotensina/análise , Núcleo Solitário/química , Nervo Vago/química , Animais , Capilares/química , Dendritos/química , Imuno-Histoquímica , Masculino , Microscopia Eletrônica , Neurônios/química , Neurônios/ultraestrutura , Terminações Pré-Sinápticas/química , Ratos , Ratos Sprague-Dawley , TransfecçãoRESUMO
Cyclooxygenase-1 (COX-1), a rate-limiting enzyme in the synthesis of prostanoids, is involved in selected vasodilatatory responses of the cerebral circulation. Cyclooxygenase-1-null mice were used to determine whether COX-1 influences cerebral ischemic damage. The middle cerebral artery was occluded in COX-1 -/- and +/+ mice (n = 9/group), and lesion volume was determined in thionin-stained sections 24 or 96 hours later. Middle cerebral artery occlusion produced larger infarcts in COX-1 -/- mice, both at 24 (35 +/- 17%; P < 0.05) and 96 hours (41 +/- 16%; P < 0.05) after ischemia. The enlargement was not due to increased susceptibility to glutamate excitotoxicity, because microinjection of N-methyl-D-aspartate or kainate in the parietal cortex produced comparable lesions in COX-1 +/+ and -/- mice ( P > 0.05; n = 8/group). To examine the contribution of hemodynamic factors to the enlargement of the infarct, cerebral blood flow was monitored by laser-Doppler flowmetry in the ischemic territory (n = 6/group). Although the reduction in cerebral blood flow was comparable in the ischemic core ( P > 0.05), at the periphery of the ischemic territory the reduction was greater in COX-1 -/- mice (-58 +/- 4%) than in COX-1 +/+ mice (-34 +/- 5%; P < 0.05). It is concluded that mice lacking COX-1 are more susceptible to focal cerebral ischemia, an effect that can be attributed to a more severe cerebral blood flow reduction in vulnerable regions at the periphery of the ischemic territory. Thus, the vascular effects of COX-1 may contribute to maintain cerebral blood flow in the postischemic brain and, as such, play a protective role in ischemic brain injury.
Assuntos
Isquemia Encefálica/genética , Isquemia Encefálica/metabolismo , Encéfalo/enzimologia , Isoenzimas/genética , Prostaglandina-Endoperóxido Sintases/genética , Animais , Encéfalo/patologia , Isquemia Encefálica/patologia , Circulação Cerebrovascular , Ciclo-Oxigenase 1 , Suscetibilidade a Doenças , Agonistas de Aminoácidos Excitatórios , Feminino , Infarto da Artéria Cerebral Média/genética , Infarto da Artéria Cerebral Média/metabolismo , Infarto da Artéria Cerebral Média/patologia , Ácido Caínico , Masculino , Proteínas de Membrana , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , N-Metilaspartato , Neurotoxinas , Lobo Parietal/patologia , Prostaglandinas/metabolismo , Acidente Vascular Cerebral/genética , Acidente Vascular Cerebral/metabolismo , Acidente Vascular Cerebral/patologiaRESUMO
Amyloid-beta (A beta)-peptides are involved in the pathophysiology of Alzheimer's dementia. We studied the effects of A beta on selected constrictor responses of cerebral circulation. Mice were anesthetized (by using urethane-chloralose) and equipped with a cranial window. Arterial pressure and blood gases were monitored and controlled. Cerebral blood flow (CBF) was monitored by a laser Doppler probe. Topical superfusion with A beta 1-40 (0.1-10 microM), but not with the reverse peptide A beta 40-1, reduced resting CBF (-29 +/- 4% at 5 microM; P < 0.05) and augmented the reduction in CBF produced by the thromboxane analog U-46619 (+45 +/- 3% at 5 microM; P < 0.05). A beta 1-40 or A beta 1-42 did not affect the reduction in CBF produced by hypocapnia. The reduction in resting CBF and the enhancement of vasoconstriction were reversed by treatment with the free radical scavengers superoxide dismutase or manganic(I-II)meso-tetrakis(4-benzoic acid)porphyrin. Substitution of the methionine residue in position 35 with norleucine, a mutation that abolishes the ability of A beta to produce free radicals, abolished its vascular effects. Nanomolar concentrations of A beta 1-40 constricted isolated pressurized middle cerebral artery segments with intrinsic tone (-16 +/- 3% at 100 nM; P < 0.05). We conclude that A beta acts directly on cerebral arteries to produce vasoconstriction and to enhance selected constrictor responses. The evidence supports the idea that A beta-induced production of reactive oxygen species plays a role in this effect. The vascular actions of A beta may contribute to the deleterious effects resulting from accumulation of this peptide in Alzheimer's dementia.
Assuntos
Peptídeos beta-Amiloides/farmacologia , Circulação Cerebrovascular/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Vasoconstrição/efeitos dos fármacos , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacologia , Doença de Alzheimer/fisiopatologia , Animais , Circulação Cerebrovascular/fisiologia , Sequestradores de Radicais Livres/farmacologia , Hipocapnia/fisiopatologia , Fluxometria por Laser-Doppler , Masculino , Metaloporfirinas/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Vasoconstrição/fisiologia , Vasoconstritores/farmacologiaRESUMO
BACKGROUND AND PURPOSE: The prostanoid-synthesizing enzyme cyclooxygenase (COX)-2 is markedly upregulated after cerebral ischemia and may participate in the mechanisms by which postischemic inflammation contributes to the late stages of ischemic brain injury. In the present study, we sought to provide additional evidence for a role of COX-2 in the mechanisms of neurotoxicity associated with inflammation. METHODS: Nine-day-old neuronal-glial cultures, prepared from the cerebral cortex of newborn C57BL/6J mice, were exposed to lipopolysaccharide (LPS), a potent proinflammatory agent. The contribution of COX-2 was investigated by using the COX-2 inhibitor NS-398. RESULTS: LPS produced a dose-dependent (0.001 to 10 microg/mL) and selective neuronal death that was well developed 72 hours after treatment. The effect was associated with a marked increase in the concentration of the COX reaction product prostaglandin E(2) (PGE(2)) and of the cytokine tumor necrosis factor-alpha (TNF-alpha). NS-398 (10 micromol/L) blocked the PGE(2) increase, attenuated the TNF-alpha increase, and prevented the neuronal death produced by LPS. TNF-alpha-blocking antibodies attenuated LPS-induced neuronal death, but the protection was less pronounced than that afforded by NS-398. LPS failed to elevate PGE(2) or to produce cell death in neuron-enriched cultures, suggesting that glial cells are required for these effects. CONCLUSIONS: COX-2, in part through TNF-alpha-related mechanisms, contributes to LPS-induced neuronal death. The data support the hypothesis that COX-2, in addition to its role in glutamate excitotoxicity, participates in the cytotoxicity associated with inflammation.
Assuntos
Inibidores de Ciclo-Oxigenase/farmacologia , Isoenzimas/antagonistas & inibidores , Lipopolissacarídeos/toxicidade , Neurônios/efeitos dos fármacos , Nitrobenzenos/farmacologia , Sulfonamidas/farmacologia , Animais , Animais Recém-Nascidos , Antígenos de Diferenciação/biossíntese , Morte Celular/efeitos dos fármacos , Células Cultivadas , Técnicas de Cocultura , Ciclo-Oxigenase 2 , Inibidores de Ciclo-Oxigenase 2 , Dinoprostona/metabolismo , Relação Dose-Resposta a Droga , Imuno-Histoquímica , Isoenzimas/metabolismo , L-Lactato Desidrogenase/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Neuroglia/citologia , Neuroglia/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Prostaglandina-Endoperóxido Sintases/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Cyclooxygenase (COX) is a prostanoid-synthesizing enzyme present in 2 isoforms: COX-1 and COX-2. Although it has long been hypothesized that prostanoids participate in cerebrovascular regulation, the lack of adequate pharmacological tools has led to conflicting results and has not permitted investigators to define the relative contribution of COX-1 and COX-2. We used the COX-1 inhibitor SC-560 and COX-1-null (COX-1(-/-)) mice to investigate whether COX-1 plays a role in cerebrovascular regulation. Mice were anesthetized (urethane and chloralose) and equipped with a cranial window. Cerebral blood flow (CBF) was measured by laser Doppler flowmetry or by the (14)C-iodoantipyrine technique with quantitative autoradiography. In wild-type mice, SC-560 (25 micromol/L) reduced resting CBF by 21+/-4% and attenuated the CBF increase produced by topical application of bradykinin (-59%) or calcium ionophore A23187 (-49%) and by systemic hypercapnia (-58%) (P<0.05 to 0.01). However, SC-560 did not reduce responses to acetylcholine or the increase in somatosensory cortex blood flow produced by vibrissal stimulation. In COX-1(-/-) mice, resting CBF assessed by (14)C-iodoantipyrine was reduced (-13% to -20%) in cerebral cortex and other telencephalic regions (P<0.05). The CBF increase produced by bradykinin, A23187, and hypercapnia, but not acetylcholine or vibrissal stimulation, were attenuated (P<0.05 to 0.01). The free radical scavenger superoxide dismutase attenuated responses to bradykinin and A23187 in wild-type mice but not in COX-1(-/-) mice, suggesting that COX-1 is the source of the reactive oxygen species known to mediate these responses. The data provide evidence for a critical role of COX-1 in maintaining resting vascular tone and in selected vasodilator responses of the cerebral microcirculation.
Assuntos
Circulação Cerebrovascular/fisiologia , Isoenzimas/metabolismo , Prostaglandina-Endoperóxido Sintases/metabolismo , Vasodilatação/fisiologia , Acetilcolina/farmacologia , Análise de Variância , Animais , Velocidade do Fluxo Sanguíneo/efeitos dos fármacos , Bradicinina/farmacologia , Calcimicina/farmacologia , Circulação Cerebrovascular/efeitos dos fármacos , Ciclo-Oxigenase 1 , Inibidores de Ciclo-Oxigenase/farmacologia , Relação Dose-Resposta a Droga , Feminino , Genótipo , Glucose/metabolismo , Hipercapnia/fisiopatologia , Isoenzimas/antagonistas & inibidores , Isoenzimas/genética , Masculino , Proteínas de Membrana , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Neocórtex/irrigação sanguínea , Penicilamina/análogos & derivados , Penicilamina/farmacologia , Prostaglandina-Endoperóxido Sintases/genética , Pirazóis/farmacologia , Superóxido Dismutase/farmacologia , Vasodilatação/efeitos dos fármacosRESUMO
Measurement of cerebral arterial and venous blood volumes during increased cerebral blood flow can provide important information regarding hemodynamic regulation under normal, pathological, and neuronally active conditions. In particular, the change in venous blood volume induced by neural activity is one critical component of the blood oxygenation level-dependent (BOLD) signal because BOLD contrast is dependent only on venous blood, not arterial blood. Thus, relative venous and arterial blood volume (rCBV) and cerebral blood flow (rCBF) in alpha-chlorolase-anesthetized rats under hypercapnia were measured by novel diffusion-weighted (19)F NMR following an i.v. administration of intravascular tracer, perfluorocarbons, and continuous arterial spin labeling methods, respectively. The relationship between rCBF and total rCBV during hypercapnia was rCBV(total) = rCBF(0.40), which is consistent with previous PET measurement in monkeys. This relationship can be linearized in a CBF range of 50-130 ml/100 g/min as DeltarCBV(total)/ DeltarCBF = 0.31 where DeltarCBV and DeltarCBF represent rCBV and rCBF changes. The average arterial volume fraction was 0.25 at a basal condition with CBF of approximately 60 ml/100 g/min and increased up to 0.4 during hypercapnia. The change in venous rCBV was 2-fold smaller than that of total rCBV (DeltarCBV(vein)/DeltarCBF = 0.15), while the arterial rCBV change was 2.5 times larger than that of total rCBV (DeltarCBV(artery)/DeltarCBF = 0.79). These NMR results were confirmed by vessel diameter measurements with in vivo videomicroscopy. The absolute venous blood volume change contributes up to 36% of the total blood volume change during hypercapnia. Our findings provide a quantitative physiological model of BOLD contrast.
Assuntos
Volume Sanguíneo , Circulação Cerebrovascular/fisiologia , Hipercapnia/fisiopatologia , Imageamento por Ressonância Magnética/métodos , Animais , Fluorocarbonos/administração & dosagem , Hemodinâmica/fisiologia , Ratos , Ratos Sprague-Dawley , Marcadores de SpinRESUMO
Cerebral ischemia is accompanied by a marked inflammatory reaction that is initiated by ischemia-induced expression of cytokines, adhesion molecules, and other inflammatory mediators, including prostanoids and nitric oxide. Preclinical studies suggest that interventions that are aimed at attenuating such inflammation reduce the progression of brain damage that occurs during the late stages of cerebral ischemia. In particular, strategies that block the activity of inflammation-related enzymes, such as inducible nitric oxide synthase and cyclo-oxygenase-2, reduce ischemic damage with an extended therapeutic window. Although a clinical trial using murine antibodies against intercellular adhesion molecule-1 did not show benefit in patients with ischemic stroke, recent data indicate that immune activation induced by the heterologous protein may have played an important role in the failure of this trial. Therefore, there is a strong rationale for continuing to explore the efficacy of anti-inflammatory therapies in the treatment of the late stages of cerebral ischemia.
Assuntos
Isquemia Encefálica/complicações , Encefalite/etiologia , Animais , Isquemia Encefálica/fisiopatologia , Ciclo-Oxigenase 2 , Citocinas/fisiologia , Encefalite/enzimologia , Encefalite/fisiopatologia , Humanos , Isoenzimas/fisiologia , Proteínas de Membrana , Óxido Nítrico/fisiologia , Óxido Nítrico Sintase/fisiologia , Prostaglandina-Endoperóxido Sintases/fisiologiaRESUMO
Cyclooxygenase-2 (COX-2), a prostanoid-synthesizing enzyme that contributes to the toxicity associated with inflammation, has recently emerged as a promising therapeutic target for several illnesses, ranging from osteoarthritis to Alzheimer's disease. Although COX-2 has also been linked to ischemic stroke, its role in the mechanisms of ischemic brain injury remains controversial. We demonstrate that COX-2-deficient mice have a significant reduction in the brain injury produced by occlusion of the middle cerebral artery. The protection can be attributed to attenuation of glutamate neurotoxicity, a critical factor in the initiation of ischemic brain injury, and to abrogation of the deleterious effects of postischemic inflammation, a process contributing to the secondary progression of the damage. Thus, COX-2 is involved in pathogenic events occurring in both the early and late stages of cerebral ischemia and may be a valuable therapeutic target for treatment of human stroke.
Assuntos
Encéfalo/patologia , Circulação Cerebrovascular/fisiologia , Ataque Isquêmico Transitório/fisiopatologia , Isoenzimas/metabolismo , N-Metilaspartato/toxicidade , Prostaglandina-Endoperóxido Sintases/metabolismo , Animais , Encéfalo/efeitos dos fármacos , Circulação Cerebrovascular/efeitos dos fármacos , Cruzamentos Genéticos , Ciclo-Oxigenase 1 , Ciclo-Oxigenase 2 , Dinoprostona/metabolismo , Lateralidade Funcional , Ácido Glutâmico/toxicidade , Heterozigoto , Homozigoto , Ataque Isquêmico Transitório/genética , Ataque Isquêmico Transitório/patologia , Ataque Isquêmico Transitório/prevenção & controle , Isoenzimas/deficiência , Isoenzimas/genética , Rim/patologia , Rim/fisiologia , Rim/fisiopatologia , Masculino , Proteínas de Membrana , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Artéria Cerebral Média , Neurotoxinas/toxicidade , Prostaglandina-Endoperóxido Sintases/deficiência , Prostaglandina-Endoperóxido Sintases/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcrição GênicaRESUMO
The assessment of cerebral interstitial oxygen tension (piO(2)) can provide valuable information regarding cerebrovascular physiology and brain function. Compartment-specific cerebral piO(2) was measured by (19)F NMR following the infusion of an oxygen-sensitive perfluorocarbon directly into the interstitial and ventricular space of the in vivo rat brain. (19)F T(1) measurements were made and cerebral piO(2) were obtained through in vitro calibrations. The effects of graded hyperoxia, hypercapnia, and hypoxia on piO(2) and cerebral blood flow (CBF) were investigated. Under normoxia (arterial pO(2) approximately 120 mm Hg), piO(2) was approximately 30 mm Hg and jugular venous pO(2) was approximately 50 mm Hg. During hyperoxia (arterial pO(2) = 90-300 mm Hg), piO(2) increased linearly with the arterial pO(2). Following hypercapnia (arterial pCO(2) = 20-60 mm Hg), the piO(2) increased sigmoidally with increasing CBF. With hypoxia (arterial pO(2) = 30-40 mm Hg), CBF increased approximately 56% and piO(2) decreased to approximately 15 mm Hg. The hypoxia-induced CBF increase was effective to some extent in compensating for the reduced piO(2). This methodology may prove useful for investigating cerebral piO(2) under pathologically or functionally altered conditions. Magn Reson Med 45:61-70, 2001.