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1.
Glia ; 68(12): 2725-2743, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32658363

RESUMO

Diabetic neuropathy has an incidence as high as 50% of diabetic patients and is characterized by damage to neurons, Schwann cells and blood vessels within the peripheral nervous system. The low-affinity neurotrophin receptor p75 (p75NTR ), particularly expressed by the Schwann cells in the peripheral nerve, has previously been reported to play a role in developmental myelination and cell survival/death. Increased levels of p75NTR , in the endoneurium and plasma from diabetic patients and rodent models of disease, have been observed, proposing that this receptor might be involved in the pathogenesis of diabetic neuropathy. Therefore, in this study, we addressed this hypothesis by utilizing a mouse model of selective nerve growth factor receptor (Ngfr) deletion in Schwann cells (SC-p75NTR -KO). Electron microscopy of sciatic nerves from mice with high fat diet induced obesity demonstrated how loss of Schwann cell-p75NTR aggravated axonal atrophy and loss of C-fibers. RNA sequencing disclosed several pre-clinical signaling alterations in the diabetic peripheral nerves, dependent on Schwann cell p75NTR signaling, specially related with lysosome, phagosome, and immune pathways. Morphological and biochemical analyses identified abundant lysosomes and autophagosomes in the C-fiber axoplasm of the diabetic SC-p75NTR -KO nerves, which together with increased Cathepsin B protein levels corroborates gene upregulation from the phagolysosomal pathways. Altogether, this study demonstrates that Schwann cell p75NTR deficiency amplifies diabetic neuropathy disease by triggering overactivation of immune-related pathways and increased lysosomal stress.


Assuntos
Neuropatias Diabéticas , Células de Schwann , Animais , Axônios , Humanos , Camundongos , Receptor de Fator de Crescimento Neural , Receptores de Fator de Crescimento Neural/genética , Nervo Isquiático
2.
Diabetologia ; 58(3): 625-32, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25523623

RESUMO

AIMS/HYPOTHESIS: We addressed the question of whether the autophagy pathway occurs in human peripheral nerves and whether this pathway is associated with peripheral neuropathy in diabetes mellitus. METHODS: By using electron microscopy, we evaluated the presence of autophagy-related structures and neuropathy in the posterior interosseous nerve of patients who had undergone carpal tunnel release and had type 1 or type 2 diabetes mellitus, and in patients with no diabetes (controls). RESULTS: Autophagy-related ultrastructures were observed in the samples taken from all patients of the three groups. The number of autophagy-associated structures was significantly higher (p < 0.05) in the nerves of patients with type 1 than type 2 diabetes. Qualitative and quantitative evaluations of fascicle area, diameter of myelinated and unmyelinated nerve fibres, the density of myelinated and unmyelinated fibres and the g-ratio of myelinated fibres were performed. We found degeneration and regeneration of a few myelinated axons in controls, and a well-developed neuropathy with the loss of large myelinated axons and the presence of many small ones in patients with diabetes. The pathology in type 1 diabetes was more extensive than in type 2 diabetes. CONCLUSIONS/INTERPRETATION: The results of this study show that the human peripheral nerves have access to the autophagy machinery, and this pathway may be regulated differently in type 1 and type 2 diabetes; insulin, presence of extensive neuropathy, and/or other factors such as duration of diabetes and HbA1c level may underlie this differential regulation.


Assuntos
Autofagia/fisiologia , Diabetes Mellitus Tipo 1/patologia , Diabetes Mellitus Tipo 2/patologia , Nervos Periféricos/patologia , Nervos Periféricos/ultraestrutura , Adulto , Idoso , Diabetes Mellitus Tipo 1/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Feminino , Humanos , Técnicas In Vitro , Masculino , Pessoa de Meia-Idade , Nervos Periféricos/metabolismo
3.
J Pept Sci ; 20(6): 429-37, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24723458

RESUMO

Human catestatin CgA352-372 (SL21) is an endogenous neuropeptide with multiple biological functions. The present study aimed to evaluate the antioxidant, antibacterial, cytotoxic, and DNA damage protective effects of SL21 neuropeptide. SL21 neuropeptide generated from the C-terminus of chromogranin A (CgA) was synthesized by solid-phase method. Synthetic peptide was subjected to various in vitro antioxidant assays including the scavenging of 1,1-diphenyl-2-pycryl-hydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(·+) ), and hydroxyl free radicals, metal ion chelation, inhibition of lipid peroxidation, and reducing power. Moreover, protective effect of SL21 on H2 O2 -induced DNA damage was analyzed using pTZ57/RT plasmid. Methylthiazoltetrazolium assay was also performed to study the cytotoxic effect of SL21 neuropeptide on human peripheral blood mononuclear cells. Furthermore, antibacterial and hemolysis assays were conducted. The results demonstrated high activities of SL21 in scavenging free radicals (DPPH, ABTS(·+) , and hydroxyl), chelating of Cu(2+) /Fe(2+) metal ions, reducing power, and inhibition of lipid peroxidation in a concentration-dependent manner. SL21 neuropeptide revealed a protective effect on DNA damage caused by hydroxyl radicals. Interestingly, the peptide exhibited no significant cytotoxicity towards peripheral blood mononuclear cells. Furthermore, SL21 peptide displayed antimicrobial activity against Staphylococcus aureus and Pseudomonas aeruginosa without any hemolytic activity on human red blood cells. Conclusively, the present study established SL21 (catestatin) as a novel antioxidative peptide that could further be investigated for its potential use as a pharmaceutical agent.


Assuntos
Antibacterianos/farmacologia , Antioxidantes/farmacologia , Cromogranina A/química , Dano ao DNA , Radicais Livres/metabolismo , Neuropeptídeos/química , Neuropeptídeos/farmacologia , Antibacterianos/síntese química , Antibacterianos/química , Antioxidantes/síntese química , Antioxidantes/química , Relação Dose-Resposta a Droga , Radicais Livres/química , Humanos , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/metabolismo , Leucócitos Mononucleares , Testes de Sensibilidade Microbiana , Neuropeptídeos/síntese química , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Relação Estrutura-Atividade
4.
Front Cell Neurosci ; 16: 856734, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35634462

RESUMO

Peripheral nerve regeneration relies on the ability of Schwann cells to support the regrowth of damaged axons. Schwann cells re-differentiate when reestablishing contact with the sprouting axons, with large fibers becoming remyelinated and small nociceptive fibers ensheathed and collected into Remak bundles. We have previously described how the receptor sortilin facilitates neurotrophin signaling in peripheral neurons via regulated trafficking of Trk receptors. This study aims to characterize the effects of sortilin deletion on nerve regeneration following sciatic crush injury. We found that Sort1 - / - mice displayed functional motor recovery like that of WT mice, with no detectable differences in relation to nerve conduction velocities and morphological aspects of myelinated fibers. In contrast, we found abnormal ensheathment of regenerated C-fibers in injured Sort1 - / - mice, demonstrating a role of sortilin for Remak bundle formation following injury. Further studies on Schwann cell signaling pathways showed a significant reduction of MAPK/ERK, RSK, and CREB phosphorylation in Sort1 - / - Schwann cells after stimulation with neurotrophin-3 (NT-3), while Schwann cell migration and myelination remained unaffected. In conclusion, our results demonstrate that loss of sortilin blunts NT-3 signaling in Schwann cells which might contribute to the impaired Remak bundle regeneration after sciatic nerve injury.

5.
Neurobiol Learn Mem ; 95(3): 270-6, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21144907

RESUMO

Alzheimer's disease (AD) is a debilitating neurodegenerative disorder characterized by increased ß-amyloid (Aß) deposition and neuronal dysfunction leading to impaired learning and recall. Ageing, heredity, and induced oxidative stress are among proposed risk factors. The increased frequency of the disease in women also suggests a role for estrogen in development of AD. In the present study, effects of the phytoestrogen genistein (10mg/kg) on learning and memory impairments was assessed in intrahippocampal Aß(1-40)-injected rats. The estrogen receptor antagonist fulvestrant was injected intracerebroventricularly in a group of Aß-lesioned rats. The Aß-injected animals exhibited the following: lower spontaneous alternation score in Y-maze tasks, impaired retention and recall capability in the passive avoidance test, and fewer correct choices and more errors in the RAM task. Genistein, but not genistein and fulvestrant, significantly improved most of these parameters. Measurements of oxidative stress markers in hippocampal tissue of Aß-injected rats showed an elevation of malondialdehyde (MDA) and nitrite content, and a reduction of superoxide dismutase (SOD) activity. Genistein significantly attenuated the increased MDA content but did not affect the nitrite content or SOD activity. These results indicate that genistein pretreatment ameliorates Aß-induced impairment of short-term spatial memory in rats through an estrogenic pathway and by inducing attenuation of oxidative stress.


Assuntos
Doença de Alzheimer/prevenção & controle , Genisteína/farmacologia , Hipocampo/efeitos dos fármacos , Aprendizagem em Labirinto/efeitos dos fármacos , Memória de Curto Prazo/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Doença de Alzheimer/induzido quimicamente , Peptídeos beta-Amiloides , Análise de Variância , Animais , Aprendizagem da Esquiva/efeitos dos fármacos , Modelos Animais de Doenças , Estradiol/administração & dosagem , Estradiol/análogos & derivados , Antagonistas de Estrogênios/administração & dosagem , Fulvestranto , Hipocampo/metabolismo , Hipocampo/fisiopatologia , Infusões Intraventriculares , Masculino , Malondialdeído/metabolismo , Memória de Curto Prazo/fisiologia , Rememoração Mental/efeitos dos fármacos , Microinjeções , Estresse Oxidativo/efeitos dos fármacos , Fragmentos de Peptídeos , Fitoestrógenos/farmacologia , Distribuição Aleatória , Ratos , Ratos Wistar , Retenção Psicológica/efeitos dos fármacos , Estatísticas não Paramétricas , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo
6.
Acta Neuropathol Commun ; 9(1): 31, 2021 02 25.
Artigo em Inglês | MEDLINE | ID: mdl-33632316

RESUMO

Pain is a common non-motor symptom of Parkinson's disease (PD), with current limited knowledge of its pathophysiology. Here, we show that peripheral inoculation of mouse alpha-synuclein (α-Syn) pre-formed fibrils, in a transgenic mouse model of PD, elicited retrograde trans-synaptic spreading of α-Syn pathology (pSer129) across sensory neurons and dorsal nerve roots, reaching central pain processing regions, including the spinal dorsal horn and the projections of the anterolateral system in the central nervous system (CNS). Pathological peripheral to CNS propagation of α-Syn aggregates along interconnected neuronal populations within sensory afferents, was concomitant with impaired nociceptive response, reflected by mechanical allodynia, reduced nerve conduction velocities (sensory and motor) and degeneration of small- and medium-sized myelinated fibers. Our findings show a link between the transneuronal propagation of α-Syn pathology with sensory neuron dysfunction and neuropathic impairment, suggesting promising avenues of investigation into the mechanisms underlying pain in PD.


Assuntos
Neuralgia/etiologia , Neuralgia/fisiopatologia , Degeneração Retrógrada/patologia , Degeneração Retrógrada/fisiopatologia , Células Receptoras Sensoriais/patologia , Sinucleinopatias/patologia , Sinucleinopatias/fisiopatologia , Animais , Modelos Animais de Doenças , Feminino , Camundongos , Camundongos Transgênicos , Microscopia Eletrônica de Transmissão , Doença de Parkinson/fisiopatologia , Células Receptoras Sensoriais/metabolismo , Células Receptoras Sensoriais/ultraestrutura , Transmissão Sináptica , alfa-Sinucleína/metabolismo
7.
Sci Rep ; 10(1): 7592, 2020 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-32371896

RESUMO

A deeper knowledge of the architecture of the peripheral nerve with three-dimensional (3D) imaging of the nerve tissue at the sub-cellular scale may contribute to unravel the pathophysiology of neuropathy. Here we demonstrate the feasibility of X-ray phase contrast holographic nanotomography to enable 3D imaging of nerves at high resolution, while covering a relatively large tissue volume. We show various subcomponents of human peripheral nerves in biopsies from patients with type 1 and 2 diabetes and in a healthy subject. Together with well-organized, parallel myelinated nerve fibres we show regenerative clusters with twisted nerve fibres, a sprouted axon from a node of Ranvier and other specific details. A novel 3D construction (with movie created) of a node of Ranvier with end segment of a degenerated axon and sprout of a regenerated one is captured. Many of these architectural elements are not described in the literature. Thus, X-ray phase contrast holographic nanotomography enables identifying specific morphological structures in 3D in peripheral nerve biopsies from a healthy subject and from patients with type 1 and 2 diabetes.


Assuntos
Neuropatias Diabéticas/diagnóstico por imagem , Neuropatias Diabéticas/patologia , Holografia , Nervos Periféricos/diagnóstico por imagem , Nervos Periféricos/patologia , Idoso , Estudos de Casos e Controles , Diabetes Mellitus Tipo 1/complicações , Diabetes Mellitus Tipo 2/complicações , Feminino , Holografia/métodos , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Microscopia , Pessoa de Meia-Idade , Nanotecnologia , Microtomografia por Raio-X/métodos
8.
Cell Biol Int ; 34(1): 61-5, 2009 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-19947934

RESUMO

In the present study, rat primary cultures were used to study the effect of lactate on the survival of hippocampal neurons in the presence or absence of glucose. Our results showed no extensive cell damage under glucose-free conditions compared with glucose-rich conditions. Addition of 10 and 50 mM lactate to glucose-free and glucose-rich media increased the cell damage significantly, as observed by morphology and lactate dehydrogenase activity. The results of the present study suggest that primary neurons in vitro are not sensitive to glucose deficiency and the presence of lactate damages the neurons in a concentration-dependent manner.


Assuntos
Hipocampo/citologia , Lactatos/toxicidade , Neurônios/metabolismo , Animais , Células Cultivadas , Embrião de Mamíferos/citologia , Glucose/farmacologia , Lactato Desidrogenases/metabolismo , Neurônios/citologia , Neurônios/efeitos dos fármacos , Ratos
9.
Front Cell Neurosci ; 13: 235, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31191256

RESUMO

Schwann cell reprogramming and differentiation are crucial prerequisites for neuronal regeneration and re-myelination to occur following injury to peripheral nerves. The neurotrophin receptor p75NTR has been identified as a positive modulator for Schwann cell myelination during development and implicated in promoting nerve regeneration after injury. However, most studies base this conclusion on results obtained from complete p75NTR knockout mouse models and cannot dissect the specific role of p75NTR expressed by Schwann cells. In this present study, a conditional knockout model selectively deleting p75NTR expression in Schwann cells was generated, where p75NTR expression is replaced with that of an mCherry reporter. Silencing of Schwann cell p75NTR expression was confirmed in the sciatic nerve in vivo and in vitro, without altering axonal expression of p75NTR. No difference in sciatic nerve myelination during development or following sciatic nerve crush injury was observed, as determined by quantification of both myelinated and unmyelinated nerve fiber densities, myelinated axonal diameter and myelin thickness. However, the absence of Schwann cell p75NTR reduced motor nerve conduction velocity after crush injury. Our data indicate that the absence of Schwann cell p75NTR expression in vivo is not critical for axonal regrowth or remyelination following sciatic nerve crush injury, but does play a key role in functional recovery. Overall, this represents the first step in redefining the role of p75NTR in the peripheral nervous system, suggesting that the Schwann cell-axon unit functions as a syncytium, with the previous published involvement of p75NTR in remyelination most likely depending on axonal/neuronal p75NTR and/or mutual glial-axonal interactions.

10.
Brain Behav ; 7(8): e00763, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28828222

RESUMO

OBJECTIVES: The progression and pathophysiology of neuropathy in impaired glucose tolerance (IGT) and type 2 diabetes (T2DM) is poorly understood, especially in relation to autophagy. This study was designed to assess whether the presence of autophagy-related structures was associated with sural nerve fiber pathology, and to investigate if endoneurial capillary pathology could predict the development of T2DM and neuropathy. PATIENTS AND METHODS: Sural nerve physiology and ultrastructural morphology were studied at baseline and 11 years later in subjects with normal glucose tolerance (NGT), IGT, and T2DM. RESULTS: Subjects with T2DM had significantly lower sural nerve amplitude compared to subjects with NGT and IGT at baseline. Myelinated and unmyelinated fiber, endoneurial capillary morphology, and the presence and distribution of autophagy structures were comparable between groups at baseline, except for a smaller myelinated axon diameter in subjects with T2DM and IGT compared to NGT. The baseline values of the subjects with NGT and IGT who converted to T2DM 11 years later demonstrated healthy smaller endoneurial capillary and higher g-ratio versus subjects who remained NGT. At follow-up, T2DM showed a reduction in nerve conduction, amplitude, myelinated fiber density, unmyelinated axon diameter, and autophagy structures in myelinated axons. Endothelial cell area and total diffusion barrier was increased versus baseline. CONCLUSIONS: We conclude that small healthy endoneurial capillary may presage the development of T2DM and neuropathy. Autophagy occurs in human sural nerves and can be affected by T2DM. Further studies are warranted to understand the role of autophagy in diabetic neuropathy.


Assuntos
Autofagia , Diabetes Mellitus Tipo 2 , Angiopatias Diabéticas , Neuropatias Diabéticas , Intolerância à Glucose/fisiopatologia , Nervo Sural/fisiopatologia , Adulto , Idoso , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/fisiopatologia , Angiopatias Diabéticas/diagnóstico , Angiopatias Diabéticas/etiologia , Angiopatias Diabéticas/fisiopatologia , Neuropatias Diabéticas/diagnóstico , Neuropatias Diabéticas/etiologia , Neuropatias Diabéticas/fisiopatologia , Feminino , Teste de Tolerância a Glucose , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Condução Nervosa
11.
J Neuropathol Exp Neurol ; 65(12): 1118-25, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17146285

RESUMO

We investigated the effects of hyperglycemia and hypoglycemia on development of peripheral neuropathy in somatic motor and sensory nerves in type 1 diabetic BB/Wor rats. The animals were maintained in a hyper- or hypoglycemic state by treatment with insulin for 3 months. Nondiabetic siblings served as controls. Qualitative analysis of the gastrocnemius and sural nerves by light and electron microscopy revealed signs of Wallerian-type axonal degeneration and regeneration of large myelinated fibers in the hypoglycemic but not the hyperglycemic animals. Degeneration was more common in the gastrocnemius nerve than in the sural nerve. In hypoglycemic rats, myelinated fibers in both the gastrocnemius and sural nerves had significantly shorter internodes and smaller diameters. The decreased fiber diameter was related (r = -0.9) to the duration of severe hypoglycemia (

Assuntos
Neuropatias Diabéticas/fisiopatologia , Hiperglicemia/fisiopatologia , Hipoglicemia/fisiopatologia , Nervos Periféricos/fisiopatologia , Degeneração Walleriana/fisiopatologia , Animais , Neuropatias Diabéticas/etiologia , Neuropatias Diabéticas/patologia , Modelos Animais de Doenças , Feminino , Hiperglicemia/complicações , Hiperglicemia/metabolismo , Hipoglicemia/complicações , Hipoglicemia/metabolismo , Microscopia Eletrônica de Transmissão , Neurônios Motores/metabolismo , Neurônios Motores/patologia , Neurônios Aferentes/metabolismo , Neurônios Aferentes/patologia , Nervos Periféricos/metabolismo , Nervos Periféricos/patologia , Ratos , Ratos Mutantes , Nervo Sural/metabolismo , Nervo Sural/patologia , Nervo Tibial/metabolismo , Nervo Tibial/patologia , Degeneração Walleriana/etiologia , Degeneração Walleriana/patologia
12.
PLoS One ; 11(9): e0163427, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27662650

RESUMO

Twenty-five percent of polyneuropathies are idiopathic. Microangiopathy has been suggested to be a possible pathogenic cause of chronic idiopathic axonal polyneuropathy (CIAP). Dysfunction of the autophagy pathway has been implicated as a marker of neurodegeneration in the central nervous system, but the autophagy process is not explored in the peripheral nervous system. In the current study, we examined the presence of microangiopathy and autophagy-related structures in sural nerve biopsies of 10 patients with CIAP, 11 controls with inflammatory neuropathy and 10 controls without sensory polyneuropathy. We did not find any significant difference in endoneurial microangiopathic markers in patients with CIAP compared to normal controls, though we did find a correlation between basal lamina area thickness and age. Unexpectedly, we found a significantly larger basal lamina area thickness in patients with vasculitic neuropathy. Furthermore, we found a significantly higher density of endoneurial autophagy-related structures, particularly in patients with CIAP but also in patients with inflammatory neuropathy, compared to normal controls. It is unclear if the alteration in the autophagy pathway is a consequence or a cause of the neuropathy. Our results do not support the hypothesis that CIAP is primarily caused by a microangiopathic process in endoneurial blood vessels in peripheral nerves. The significantly higher density of autophagy structures in sural nerves obtained from patients with CIAP and inflammatory neuropathy vs. controls indicates the involvement of this pathway in neuropathy, particularly in CIAP, since the increase in density of autophagy-related structures was more pronounced in patients with CIAP than those with inflammatory neuropathy. To our knowledge this is the first report investigating signs of autophagy process in peripheral nerves in patients with CIAP and inflammatory neuropathy.

13.
Diabetes Technol Ther ; 7(3): 509-15, 2005 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15929682

RESUMO

BACKGROUND: The aim of this study was to ascertain whether dysglycemic episodes occur in institutionalized elderly persons and, if that is the case, to determine whether such episodes are related to meal patterns. Another objective was to investigate the feasibility of subcutaneous (s.c.) glucose measurements in the elderly using a Medtronic MiniMed (Sylmar, CA) continuous glucose monitoring system (CGMS). METHODS: Nine nursing home residents (74-95 years old) without known diabetes or other metabolic disorders were included. The s.c. glucose level was measured for 3 days with the Medtronic MiniMed CGMS. Capillary blood glucose was measured four times daily with a Glucometer Elite device (Bayer, Leverkusen, Germany). Body mass index and basal metabolic rate were calculated, and food intake was recorded. RESULTS: The s.c. glucose level fluctuated noticeably over time; 22.5% of the values recorded during the 3-day period were > or = 8 mmol/L, and values < 3.5 mmol/L were rarely seen. A marked (> 5 mmol/L) and short-term (2-4 h) increase in s.c. glucose was seen after a meal. The mean capillary blood glucose concentration was 7.5 +/- 1.8 mmol/L. Capillary blood glucose > or = 8 mmol/L was recorded on 32.5% of the measurement occasions, and no values were < 3.5 mmol/L. The s.c. glucose values agreed with corresponding capillary blood glucose levels (mean r = 0.75; range 0.43-0.86). Five participants consumed less energy than recommended according to their age, weight, and physical activity level. CONCLUSIONS: Postprandial hyperglycemia frequently occurs in elderly people living in nursing homes. The CGMS is convenient to use to detect hyperglycemia in this age group.


Assuntos
Glicemia/metabolismo , Hiperglicemia/epidemiologia , Monitorização Ambulatorial/métodos , Período Pós-Prandial , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , Registros de Dieta , Feminino , Instituição de Longa Permanência para Idosos , Humanos , Hiperglicemia/diagnóstico , Consentimento Livre e Esclarecido , Masculino , Casas de Saúde , Reprodutibilidade dos Testes
14.
J Vis Exp ; (106): e53113, 2015 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-26709729

RESUMO

As glial cells in the brain, astrocytes have diverse functional roles in the central nervous system. In the presence of harmful stimuli, astrocytes modify their functional and structural properties, a condition called reactive astrogliosis. Here, a protocol for assessment of the morphological properties of astrocytes is presented. This protocol includes quantification of 12 different parameters i.e. the surface area and volume of the tissue covered by an astrocyte (astrocyte territory), the entire astrocyte including branches, cell body, and nucleus, as well as total length and number of branches, the intensity of fluorescence immunoreactivity of antibodies used for astrocyte detection, and astrocyte density (number/1,000 µm(2)). For this purpose three-dimensional (3D) confocal microscopic images were created, and 3D image analysis software such as Volocity 6.3 was used for measurements. Rat brain tissue exposed to amyloid beta1-40 (Aß1-40) with or without a therapeutic intervention was used to present the method. This protocol can also be used for 3D morphometric analysis of other cells from either in vivo or in vitro conditions.


Assuntos
Astrócitos/citologia , Microscopia Confocal/métodos , Peptídeos beta-Amiloides/farmacologia , Animais , Encéfalo/citologia , Encéfalo/efeitos dos fármacos , Imageamento Tridimensional/métodos , Fragmentos de Peptídeos/farmacologia , Ratos
15.
Eur J Pharmacol ; 764: 195-201, 2015 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-26148826

RESUMO

Alzheimer's disease (AD) is one of the prevalent neurological disorders of the central nervous system hallmarked by increased beta-amyloid (Aß) deposition and ensuing learning and memory deficit. In the present study, the beneficial effect of naringenin on improvement of learning and memory was evaluated in an Alzheimer's disease rat model. The Aß-injected rats showed a lower alternation score in Y-maze task, impairment of retention and recall capability in passive avoidance test, and lower correct choices and higher errors in radial arm maze (RAM) task as compared to sham group in addition to enhanced oxidative stress and apoptosis. Naringenin, but not a combination of naringenin and fulvestrant (an estrogenic receptor antagonist) significantly improved the performance of Aß-injected rats in passive avoidance and RAM tasks. Naringenin pretreatment of Aß-injected rats also lowered hippocampal malondialdehyde (MDA) with no significant effect on nitrite and superoxide dismutase (SOD) activity in addition to lowering apoptosis. These results suggest naringenin pretreatment attenuates Aß-induced impairment of learning and memory through mitigation of lipid peroxidation and apoptosis and its beneficial effect is somewhat mediated via estrogenic pathway.


Assuntos
Doença de Alzheimer/tratamento farmacológico , Flavanonas/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Doença de Alzheimer/metabolismo , Animais , Apoptose/efeitos dos fármacos , Aprendizagem da Esquiva/efeitos dos fármacos , Fragmentação do DNA/efeitos dos fármacos , Modelos Animais de Doenças , Flavanonas/farmacologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , Malondialdeído/metabolismo , Aprendizagem em Labirinto/efeitos dos fármacos , Memória/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Nitritos/metabolismo , Ratos Wistar , Superóxido Dismutase/metabolismo
16.
Diabetes Technol Ther ; 4(3): 305-12, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12165169

RESUMO

Laboratory animals are extensively used in diabetic research. However, it is not known whether the glucose dynamics in laboratory animals are similar to the dynamics in humans. The aim of the present study is to see whether the Medtronic MiniMed continuous subcutaneous glucose monitoring system can be used to record fluctuations of the glucose level in freely moving normal and insulin-treated diabetic rats. The monitoring system was applied during 3 days to normal and diabetic hyperglycemic and hypoglycemic rats treated with insulin implants. Corresponding data from type 1 diabetic patients with poor glycemic control were selected retrospectively in order to note the similarities and differences. In normal rats the subcutaneous glucose level varied slightly (median = 111 mg/dL). In hyperglycemic rats the subcutaneous glucose values fluctuated markedly around a median of 226 mg/dL. The fluctuations formed a short-wave pattern with a low amplitude, superimposed on a long-wave pattern with a high amplitude. The subcutaneous glucose profile seen in type 1 diabetic patients (median = 180 mg/dL) was similar to that observed in hyperglycemic rats. In hypoglycemic rats, the subcutaneous glucose level fluctuated moderately around a median of 55 mg/dL. In these rats the fluctuations formed a short-wave pattern with low amplitude, without any obvious long-wave pattern. The subcutaneous glucose values conformed to corresponding blood glucose measurements. We conclude that the Medtronic MiniMed continuous glucose monitoring system can be used to record the subcutaneous glucose level over time in freely moving rats.


Assuntos
Glicemia/análise , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Tipo 1/sangue , Monitorização Ambulatorial/métodos , Adolescente , Animais , Automonitorização da Glicemia , Calibragem , Criança , Diabetes Mellitus Experimental/tratamento farmacológico , Feminino , Hemoglobinas Glicadas/análise , Humanos , Hipoglicemia/sangue , Insulina/uso terapêutico , Masculino , Miniaturização , Monitorização Ambulatorial/instrumentação , Monitorização Ambulatorial/normas , Ratos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Pele/irrigação sanguínea , Fatores de Tempo
17.
Handb Clin Neurol ; 126: 513-32, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25410242

RESUMO

Hypoglycemia occurs in diabetic patients as a consequence of treatment with hypoglycemic agents, in insulinoma patients as a result of excessive insulin production, and in infants as a result of abnormal regulation of metabolism. Profound hypoglycemia can cause structural and functional disturbances in both the central (CNS) and the peripheral nervous system (PNS). The brain is damaged by a short and severe episode of hypoglycemia, whereas PNS pathology appears after a mild and prolonged episode. In the CNS, damaged mitochondria, elevated intracellular Ca2(+) level, released cytochrome c to the cytosol, extensive production of superoxide, increased caspase-3 activity, release of aspartate and glutamate from presynaptic terminals, and altered biosynthetic machinery can lead to neuronal cell death in the brain. Considering the PNS, chronic hypoglycemia is associated with delayed motor and sensory conduction velocities in peripheral nerves. With respect to pathology, hypoglycemic neuropathy in the PNS is characterized by Wallerian-like axonal degeneration that starts at the nerve terminal and progresses to a more proximal part of the axon, and motor axons to the muscles may be more severely damaged than sensory axons. Since excitatory neurotransmitters primarily involve the neuron in the CNS, this "dying back" pattern of axonal damage in the PNS may involve mechanisms other than excitotoxicity.


Assuntos
Diabetes Mellitus/diagnóstico , Hipoglicemia/induzido quimicamente , Hipoglicemia/diagnóstico , Hipoglicemiantes/efeitos adversos , Doenças do Sistema Nervoso/induzido quimicamente , Doenças do Sistema Nervoso/diagnóstico , Animais , Diabetes Mellitus/tratamento farmacológico , Diabetes Mellitus/epidemiologia , Humanos , Hipoglicemia/epidemiologia , Doenças do Sistema Nervoso/epidemiologia
18.
Braz J Microbiol ; 45(4): 1393-9, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25763046

RESUMO

So far, the antibacterial activity of some organic and inorganic compounds has been studied. Barium zirconate titanate [Ba(ZrxTi1-x)O3] (x = 0.05) nanoparticle is an example of inorganic materials. In vitro studies have provided evidence for the antibacterial activity of this nanoparticle. In the current study, the nano-powder was synthesized by sol-gel method. X-ray diffraction showed that the powder was single-phase and had a perovskite structure at the calcination temperature of 1000 °C. Antibacterial activity of the desired nanoparticle was assessed on two gram-positive (Staphylococcus aureus PTCC1431 and Micrococcus luteus PTCC1625) and two gram-negative (Escherichia coli HP101BA 7601c and clinically isolated Klebsiella pneumoniae) bacteria according to Radial Diffusion Assay (RDA). The results showed that the antibacterial activity of BZT nano-powder on both gram-positive and gram-negative bacteria was acceptable. The minimum inhibitory concentration of this nano-powder was determined. The results showed that MIC values for E. coli, K. pneumoniae, M. luteus and S. aureus were about 2.3 µg/mL, 7.3 µg/mL, 3 µg/mL and 12 µg/mL, respectively. Minimum bactericidal concentration (MBC) was also evaluated and showed that the growth of E. coli, K. pneumoniae, M. luteus and S. aureus could be decreased at 2.3, 14, 3 and 18 µg/mL of BZT. Average log reduction in viable bacteria count in time-kill assay ranged between 6 Log10 cfu/mL to zero after 24 h of incubation with BZT nanoparticle.


Assuntos
Antibacterianos/farmacologia , Compostos de Bário/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Nanopartículas/química , Titânio/farmacologia , Zircônio/farmacologia , Compostos de Bário/síntese química , Compostos de Bário/química , Testes de Sensibilidade Microbiana , Difração de Raios X
19.
PLoS One ; 8(10): e76526, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24130779

RESUMO

Astrocytes are highly involved in regulation and homeostasis of the extracellular environment in the healthy brain. In pathological conditions, these cells play a major role in the inflammatory response seen in CNS tissues, which is called reactive astrogliosis and includes hypertrophy and proliferation of astrocytes. Here, we performed 3D confocal microscopy to evaluate the morphological response of reactive astrocytes positive for glial fibrillary acidic protein (GFAP) in rats, to the presence of Aß(1-40) in the rat brain before and after treatment with genistein. In 50 astrocytes per animal, we measured the volume and surface area for the nucleus, cell body, the entire cell, the tissue covered by single astrocytes and quantified the number and length of branches, the density of the astrocytes and the intensity of GFAP immunoreactivity. Injecting Aß(1-40) into the brain of rats caused astrogliosis indicated by increased values for all measured parameters. Mass spectrometric analysis of hippocampal tissue in Aß(1-40)-injected brain showed decreased amounts of tubulins, enolases and myelin basic protein, and increased amounts of dihydropyrimidinase-related protein 2. In Aß(1-40)-injected rats pretreated with genistein, GFAP intensity was decreased to the sham-operated group level, and Aß(1-40)-induced astrogliosis was significantly ameliorated.


Assuntos
Peptídeos beta-Amiloides/farmacologia , Genisteína/farmacologia , Genisteína/uso terapêutico , Gliose/tratamento farmacológico , Gliose/patologia , Imageamento Tridimensional , Fragmentos de Peptídeos/farmacologia , Proteoma/efeitos dos fármacos , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Astrócitos/patologia , Contagem de Células , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Tamanho Celular/efeitos dos fármacos , Proteína Glial Fibrilar Ácida/metabolismo , Gliose/induzido quimicamente , Gliose/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , Microscopia Confocal , Proteoma/metabolismo , Proteômica , Ratos , Ratos Wistar
20.
Brain Res ; 1429: 145-54, 2012 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-22079317

RESUMO

We addressed the question of whether injection of Amyloid beta (Aß)(1-40) in the rat brain is associated with pathology in the hippocampus, and if genistein has any protective effect against the neuronal damage caused by Aß(1-40). Genistein is a plant-derived compound with a structure similar to that of the female sex hormone estrogen and it was recently shown that pretreatment with a single dose of genistein ameliorated learning and memory deficits in an (Aß)(1-40) rat model of Alzheimer's disease. Here, we report that injection of the amyloid peptide into the hippocampus of rats led to formation of Aß(1-40) positive aggregates close to the lateral blade of the dentate gyrus (DGlb). We also observed the following in the hippocampus: extensive cell death in the DGlb (P<0.0001), CA1 (P=0.03), and CA3 (P=0.002); an increased number of iNOS-expressing cells (P=0.01) and gliosis. Genistein given to rats by gavage 1h before injection of Aß(1-40) inhibited the formation of Aß(1-40) positive aggregates in the brain tissue and led to increased number of nNOS(+) (P=0.0001) cells in the hippocampus compared to sham-operated genistein-treated controls. Treatment with genistein also alleviated the extensive astrogliosis that occurred in Aß(1-40)-injected hippocampus to a level similar to that observed in sham-operated rats. We conclude that the neurons in the DGlb are most sensitive to Aß(1-40), and a single dose of genistein can ameliorate Aß(1-40) induced pathology.


Assuntos
Peptídeos beta-Amiloides/toxicidade , Genisteína/farmacologia , Hipocampo/efeitos dos fármacos , Hipocampo/patologia , Fragmentos de Peptídeos/toxicidade , Fitoestrógenos/farmacologia , Animais , Gliose/induzido quimicamente , Gliose/patologia , Imuno-Histoquímica , Masculino , Ratos , Ratos Wistar
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