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1.
Science ; 369(6507)2020 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-32855309

RESUMO

Neuronal synapses undergo structural and functional changes throughout life, which are essential for nervous system physiology. However, these changes may also perturb the excitatory-inhibitory neurotransmission balance and trigger neuropsychiatric and neurological disorders. Molecular tools to restore this balance are highly desirable. Here, we designed and characterized CPTX, a synthetic synaptic organizer combining structural elements from cerebellin-1 and neuronal pentraxin-1. CPTX can interact with presynaptic neurexins and postsynaptic AMPA-type ionotropic glutamate receptors and induced the formation of excitatory synapses both in vitro and in vivo. CPTX restored synaptic functions, motor coordination, spatial and contextual memories, and locomotion in mouse models for cerebellar ataxia, Alzheimer's disease, and spinal cord injury, respectively. Thus, CPTX represents a prototype for structure-guided biologics that can efficiently repair or remodel neuronal circuits.


Assuntos
Proteína C-Reativa/farmacologia , Proteínas do Tecido Nervoso/farmacologia , Vias Neurais/efeitos dos fármacos , Precursores de Proteínas/farmacologia , Receptores de AMPA/metabolismo , Proteínas Recombinantes/farmacologia , Sinapses/efeitos dos fármacos , Doença de Alzheimer/terapia , Animais , Proteína C-Reativa/química , Proteína C-Reativa/uso terapêutico , Ataxia Cerebelar/terapia , Modelos Animais de Doenças , Células HEK293 , Hipocampo , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/uso terapêutico , Domínios Proteicos , Precursores de Proteínas/química , Precursores de Proteínas/uso terapêutico , Receptores de Glutamato/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/uso terapêutico , Coluna Vertebral/efeitos dos fármacos , Coluna Vertebral/fisiologia
2.
ScientificWorldJournal ; 2020: 7910702, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32148468

RESUMO

Dye-Sensitized Solar Cell (DSSC) constitutes a solar cell using natural dyes from plants that are adsorbed in semiconductors to convert solar energy into electrical energy. DSSC has relatively inexpensive fabrication costs, is easy to produce, works in visible light, and is environmentally friendly. The disadvantage of DSSC is that its efficiency is still low compared to silicon solar cells. This low efficiency is due to obstacles in the flow of electric current on DSSC. In this study, DSSC has been successfully fabricated with the deposition of clathrin protein from cow brain. The zwitterions effect of protein on cow brain is able to reduce resistance and increase electric current on DSSC. The zwitterions effect of cow brain protein that fills gaps or empty spaces between TiO2 particles generates acidic reactions (capturing electrons) and bases (releasing electrons); hence, proteins in the cow brain are able to function as electron bridges between TiO2 molecules and generate an increase in electric current in DSSC. The method used in this research was to deposit clathrin protein from cow brain in a porous TiO2 semiconductor with a concentration of 0%, 25%, 50%, and 75%. Tests carried out on DSSC that have been performed were X-Ray Diffractometer (XRD) testing to determine the crystal structure formed, Fourier Transform Infrared Spectroscopy (FTIR) testing to determine the functional groups formed on DSSC, Scanning Electron Microscopy (SEM) testing to determine the surface morphological characteristics of the DSSC layer, and testing the efficiency using AM 1.5 G solar simulator (1000 W/m2) to determine the efficiency changes that occur in DSSC. From the XRD test results by increasing the concentration of cow brain protein in DSSC, the structure of amino acid crystals also increased and the crystal size increased with the largest crystal size of 42.25 nm at the addition of 75% of cow brain protein. FTIR test results show that the addition of cow brain protein will form functional protein-forming amino groups on DSSC. FTIR analysis shows the sharp absorption of energy by protein functional groups in the FTIR spectrum with increasing concentration of cow brain protein in DSSC. The SEM test results show that the concentration of additional molecules of protein deposited into TiO2 increases and the cavity or pore between the TiO2 molecules decreases. The reduction of cavities in the layers indicates that protein molecules fill cavities that exist between TiO2 molecules. From the results of testing using AM 1.5 G solar simulator (1000 W/m2), the highest efficiency value is 1.465% with the addition of 75% brain protein concentration.


Assuntos
Encéfalo , Corantes , Proteínas do Tecido Nervoso/farmacologia , Compostos de Amônio Quaternário/farmacologia , Energia Solar , Algoritmos , Animais , Encéfalo/metabolismo , Bovinos , Corantes/metabolismo , Fontes de Energia Elétrica , Modelos Teóricos , Proteínas do Tecido Nervoso/química , Compostos de Amônio Quaternário/química , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
3.
F1000Res ; 82019.
Artigo em Inglês | MEDLINE | ID: mdl-31583078

RESUMO

Complex mechanisms control the signaling of neurotrophins through p75 NTR and Trk receptors, allowing cellular responses that are highly context dependent, particularly in the nervous system and particularly with regard to the neurotrophin brain-derived neurotrophic factor (BDNF). Recent reports describe a variety of sophisticated regulatory mechanisms that contribute to such functional flexibility. Mechanisms described include regulation of trafficking of alternative BDNF transcripts, regulation of post-translational processing and secretion of BDNF, engagement of co-receptors that influence localization and signaling of p75 NTR and Trk receptors, and control of trafficking of receptors in the endocytic pathway and during anterograde and retrograde axonal transport.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/fisiologia , Transdução de Sinais , Transporte Axonal , Endocitose , Humanos , Proteínas do Tecido Nervoso/farmacologia , Processamento de Proteína Pós-Traducional , Receptores de Fator de Crescimento Neural/fisiologia
4.
In Vitro Cell Dev Biol Anim ; 55(9): 756-765, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31420803

RESUMO

Vascular endothelial growth factor (VEGF) inhibition forms the basis for anti-angiogenic therapies. With the methods based on the monoclonal antibody-mediated typical VEGF blockade, pathological angiogenesis in the tumor microenvironment is inhibited and the limitation of tumor growth is provided; however, the existing tumor tissue cannot be intervened. In this study, the anti-angiogenic effects of Semaphorin (SEMA) 3F, which has frequently been reported to have tumor suppressive properties, on a chick chorioallantoic membrane model as well as in vitro cell-cell interactions were investigated and comparatively assessed using anti-VEGF antibody. Vascular endothelial cells and chick embryos were stimulated with 10-16 ng/mL VEGF165 prior to SEMA 3F administration in order to generate pathological vascularization conditions. Both in vitro and in ovo results revealed that SEMA 3F suppressed VEGF165-induced abnormal vascularization more effectively than anti-VEGF. Moreover, the required dose of SEMA 3F was significantly lower than that of anti-VEGF (103 times less under in ovo conditions). In light of these results, SEMA 3F is recommended as an important therapeutic agent for the prevention of pathological angiogenesis. SEMA 3F may offer an effective and efficient anti-angiogenic intervention that can be administered at a lower dose alternative to typical VEGF blocking agents.


Assuntos
Proteínas de Membrana/genética , Neoplasias/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Proteínas do Tecido Nervoso/genética , Fator A de Crescimento do Endotélio Vascular/genética , Animais , Anticorpos Monoclonais/farmacologia , Embrião de Galinha , Membrana Corioalantoide/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Proteínas de Membrana/farmacologia , Morfogênese/efeitos dos fármacos , Neoplasias/genética , Neoplasias/imunologia , Neoplasias/patologia , Neovascularização Patológica/genética , Neovascularização Patológica/imunologia , Neovascularização Patológica/patologia , Proteínas do Tecido Nervoso/farmacologia , Microambiente Tumoral/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/farmacologia
5.
Cells ; 8(8)2019 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-31362466

RESUMO

NK cells (natural killer cells) being a part of the innate immune system have been shown to be involved in immunoregulation of autoimmune diseases. Previously we have shown that HINT1/Hsp70 treatment induced regulatory NK cells ameliorating experimental autoimmune encephalomyelitis (EAE) course and CD4+ T cells proliferation. NK cells were isolated from mice treated with HINT1/Hsp70 and co-cultured with proteolipid protein (PLP)-stimulated CD4+ T cells isolated from EAE mice. Cell proliferation was assessed by thymidine uptake, cytotoxicity by lactate dehydrogenase (LDH) release assay and fluorescence activated cell sorting (FACS) analysis, protein expression by Western blot, mRNA by quantitative RT-PCR. Gene related to anergy in lymphocytes (GRAIL) expression was downregulated by specific siRNA and GRAIL overexpression was induced by pcDNA-GRAIL transfection. HINT1/Hsp70 pretreatment of EAE SJL/J mice ameliorated EAE course, suppressed PLP-induced T cell proliferation by enhancing T cell expression of GRAIL as GRAIL downregulation restored T cell proliferation. HINT1/Hsp70 treatment induced immunoregulatory NK cells which inhibited PLP-stimulated T cell proliferation not depending on T cell necrosis and apoptosis. This immunoregulatory NK cell function depended on NK cell expression of GRAIL as GRAIL downregulation diminished inhibition of NK cell suppression of T cell proliferation. Similarly GRAIL overexpression in NK cells induced their regulatory function. HINT1/Hsp70 treatment generated regulatory NK cells characterized by expression of GRAIL.


Assuntos
Linfócitos T CD4-Positivos/citologia , Encefalomielite Autoimune Experimental/imunologia , Proteínas de Choque Térmico HSP70/administração & dosagem , Células Matadoras Naturais/citologia , Proteínas do Tecido Nervoso/administração & dosagem , Ubiquitina-Proteína Ligases/genética , Animais , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Anergia Clonal , Técnicas de Cocultura , Encefalomielite Autoimune Experimental/etiologia , Feminino , Proteínas de Choque Térmico HSP70/farmacologia , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/metabolismo , Camundongos , Proteína Proteolipídica de Mielina/efeitos adversos , Proteína Proteolipídica de Mielina/metabolismo , Proteínas do Tecido Nervoso/farmacologia , Proteolipídeos/farmacologia , Ubiquitina-Proteína Ligases/metabolismo
6.
Pharmacol Biochem Behav ; 181: 101-109, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31054945

RESUMO

While insulin secreted from pancreas plays a pivotal role in the control of glucose homeostasis, it also interacts with hypothalamic sites and negatively influences the energy balance. The present study was undertaken to reveal the functional interaction between cocaine- and amphetamine-regulated transcript (CART), a well-known anorexic peptide, and insulin within the framework of hypothalamus in the regulation of feeding behavior and body weight. Insulin was administered daily by intracerebroventricular (icv) route, alone or in combination with CART (icv) for a period of seven days. Immediately thereafter, preweighed food was offered to the animals at the commencement of the dark phase. The food intake and body weight were measured daily just prior to next injection. Furthermore, brains of insulin-treated rats were processed for the immunohistochemical analysis of CART-containing elements in the hypothalamus. Treatment with insulin (6 mU, icv) for a period of 7 days caused a significant decrease in food intake and body weight as compared to control. Concomitant administration of CART (0.5 µg, icv) potentiated insulin-induced anorexia and weight loss. Insulin administration resulted in a significant increase in CART immunoreactivity in the hypothalamic arcuate, paraventricular, dorsomedial and ventromedial nuclei. We suggest that increased CART contents in the hypothalamus may be causally linked with anorexia and weight loss induced by insulin.


Assuntos
Peso Corporal/efeitos dos fármacos , Comportamento Alimentar/efeitos dos fármacos , Insulina/farmacologia , Proteínas do Tecido Nervoso/imunologia , Proteínas do Tecido Nervoso/farmacologia , Neuropeptídeos/imunologia , Neuropeptídeos/farmacologia , Animais , Anorexia/induzido quimicamente , Anticorpos Monoclonais/farmacologia , Ingestão de Alimentos/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Hipotálamo/imunologia , Imuno-Histoquímica , Insulina/administração & dosagem , Masculino , Proteínas do Tecido Nervoso/administração & dosagem , Neuropeptídeos/administração & dosagem , Fotoperíodo , Ratos , Ratos Sprague-Dawley , Perda de Peso/efeitos dos fármacos
7.
Microb Pathog ; 133: 103546, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31112769

RESUMO

With this study, we investigated the effect of synthetic antimicrobial peptides Pep19-2.5 and Pep194LF alone or in combination with antibiotics on S. mutans growth and biofilm formation/disruption. We also examined the cytotoxic effect of each peptide on monocytes. S. mutans was cultured in the presence of different concentrations of each peptide. We showed that Pep19-2.5 and Pep19-4LF were able to significantly (p ≤ 0.01) inhibit the growth of S. mutans. The synthetic peptides also decreased biofilm formation by S. mutans. Furthermore, both peptides reduced the viability of S. mutans in already formed biofilms. The combination of each peptide with antibiotics (penicillin/streptomycin, P/S) produced additive interactions which inhibited S. mutans growth and biofilm formation. Pep19-2.5 and Pep19-4LF were nontoxic, as they did not decrease monocyte viability and did not increase the lactate dehydrogenase activity of the exposed cells. In conclusion, synthetic peptides Pep19-2.5 and Pep19-4LF did inhibit S. mutans growth and its capacity to form biofilm. Both peptides were found to be nontoxic to monocytes. These data provide new insight into the efficacy of synthetic peptides Pep19-2.5 and Pep19-4LF against S. mutans. These peptides may thus be useful in controlling the adverse effects of this cariogenic bacterium in human.


Assuntos
Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Proteínas do Tecido Nervoso/farmacologia , Peptídeos/farmacologia , Streptococcus mutans/efeitos dos fármacos , Peptídeos Catiônicos Antimicrobianos , Biofilmes/crescimento & desenvolvimento , Sobrevivência Celular/efeitos dos fármacos , Combinação de Medicamentos , Sinergismo Farmacológico , Humanos , L-Lactato Desidrogenase/metabolismo , Testes de Sensibilidade Microbiana , Monócitos/efeitos dos fármacos , Proteínas do Tecido Nervoso/síntese química , Penicilinas/farmacologia , Peptídeos/síntese química , Streptococcus mutans/crescimento & desenvolvimento
8.
Acta Biomater ; 90: 412-423, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30951897

RESUMO

Intracellular transport is fundamental for neuronal function and development and is dependent on the formation of stable actin filaments. N-cadherin, a cell-cell adhesion protein, is actively involved in neuronal growth and actin cytoskeleton organization. Various groups have explored how neurons behaved on substrates engineered to present N-cadherin; however, few efforts have been made to examine how these surfaces modulate neuronal intracellular transport. To address this issue, we assembled a substrate to which recombinant N-cadherin molecules are physiosorbed using graphene oxide (GO) or reduced graphene oxide (rGO). N-cadherin physisorbed on GO and rGO led to a substantial enhancement of intracellular mass transport along neurites relative to N-cadherin on glass, due to increased neuronal adhesion, neurite extensions, dendritic arborization and glial cell adhesion. This study will be broadly useful for recreating active neural tissues in vitro and for improving our understanding of the development, homeostasis, and physiology of neurons. STATEMENT OF SIGNIFICANCE: Intracellular transport of proteins and chemical cues is extremely important for culturing neurons in vitro, as they replenish materials within and facilitate communication between neurons. Various studies have shown that intracellular transport is dependent on the formation of stable actin filaments. However, the extent to which cadherin-mediated cell-cell adhesion modulates intracellular transport is not heavily explored. In this study, N-cadherin was adsorbed onto graphene oxide-based substrates to understand the role of cadherin at a molecular level and the intracellular transport within cells was examined using spatial light interference microscopy. As such, the results of this study will serve to better understand and harness the role of cell-cell adhesion in neuron development and regeneration.


Assuntos
Caderinas , Grafite , Proteínas do Tecido Nervoso , Neuritos/metabolismo , Neurogênese/efeitos dos fármacos , Animais , Transporte Biológico Ativo/efeitos dos fármacos , Caderinas/química , Caderinas/farmacologia , Adesão Celular/efeitos dos fármacos , Grafite/química , Grafite/farmacologia , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/farmacologia , Ratos , Ratos Long-Evans
9.
Life Sci ; 222: 29-35, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30826495

RESUMO

Recently, several plexins and semaphorins have been associated with osteoclastogenesis, a vital process for bone remodeling. Plexin-A2 is implicated in bone homeostasis, however, whether it plays a role in osteoclastogenesis and the underlying mechanism remain unknown. We show that plexin-A2 expression is upregulated during RANKL-induced osteoclastogenesis. In addition, the soluble Sema6A fused with IgG1 Fc region (Fc-Sema6A) interacts with plexin-A2 from cell lysates of osteoclasts, suggesting that plexin-A2 acts as a receptor of Sema6A in osteoclasts. Moreover, Sema6A treatment stimulates RANKL-induced osteoclastogenesis, and this effect is abolished when plexin-A2 is neutralized, which illustrates an indispensable role of plexin-A2 in mediating Sema6A effect on osteoclastogenesis. Mechanistically, Sema6A-plexin-A2 axis enhances RANKL-induced activation of PLCγ as well as downstream target NFATc1, one master transcriptional factor of osteoclastogenesis. Lastly, inhibition of PLCγ by pharmacological inhibitor U73122 abrogates Sema6A-stimulated NFATc1 activation and RANKL-induced osteoclastogenesis, thus demonstrating that the PLCγ-mediated NFATc1 activation accounts for the promotive role of Sema6A-plexin-A2 axis in RANKL-induced osteoclastogenesis. Taken together, this study uncovers a novel role of Sema6A and plexin-A2 in osteoclastogenesis, and also offers them as possible therapeutic targets in the intervention of osteolytic diseases.


Assuntos
Fatores de Transcrição NFATC/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Osteogênese/fisiologia , Fosfolipase C gama/metabolismo , Ligante RANK/metabolismo , Receptores de Superfície Celular/metabolismo , Semaforinas/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Células Cultivadas , Estrenos/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Proteínas do Tecido Nervoso/farmacologia , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Osteogênese/efeitos dos fármacos , Inibidores de Fosfodiesterase/farmacologia , Fosfolipase C gama/antagonistas & inibidores , Pirrolidinonas/farmacologia , Semaforinas/farmacologia
10.
Anticancer Res ; 39(3): 1267-1273, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30842157

RESUMO

BACKGROUND: The members of the slit homolog (SLIT) and roundabout homolog (ROBO) families have emerged as important signaling molecules in tumor metastasis. This study analyzed their role in regulating breast cancer (BC) cell motility and chemotaxis and assessed expression of ROBO1 in brain metastases (BMs) of breast, lung, and colon cancer, and in peritoneal metastases (PMs) of ovarian cancer. MATERIALS AND METHODS: The BC cell line MDA-MB231 was subjected to scratch, motility, and chemotaxis assays using heparin and a purified recombinant N-terminal SLIT2 fragment. Protein expression was assessed in primary tumors and metastases by immunohistochemistry. RESULTS: Exposure to SLIT2 induced MDA-MB231 cell motility, but no significant chemotaxis without the presence of heparin. ROBO1 was expressed in 4/5 primary BC and in 18/21 BC-derived BM samples; 7/9 BM primary lung cancer samples also stained positive. In contrast, BMs from colorectal cancer were negative for ROBO1. Primary ovarian cancer and ovarian PM showed ROBO1 expression in 0/6 and in only 2/6 samples, respectively, whereas SLIT2 was observed in 1/6 primary cancer and in 6/6 PMs samples. CONCLUSION: SLIT2 can induce BC cell motility and chemotaxis, but the latter requires the presence of heparin. BM expression of ROBO1 is a common feature of some, but not all cancer types. SLIT2 expression appears to be a general feature of ovarian cancer-derived PMs.


Assuntos
Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Receptores Imunológicos/metabolismo , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/secundário , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Feminino , Heparitina Sulfato/farmacologia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Proteínas do Tecido Nervoso/farmacologia , Neoplasias Peritoneais/metabolismo , Neoplasias Peritoneais/secundário
11.
BMC Neurosci ; 20(1): 11, 2019 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-30885137

RESUMO

BACKGROUND: Pre-treatment with the corticotropin-releasing factor antagonist α-helical CRF9-41 prevents inhibition of gastric emptying by cocaine-and amphetamine-regulated transcript peptide at a dorsal hindbrain level, but its inhibition of sucrose intake is not affected. This is suggestive of separable underlying mechanisms of action in the caudal brainstem for cocaine-and amphetamine-regulated transcript peptide with regard to food intake and gastrointestinal functions. Here we further examine cocaine-and amphetamine-regulated transcript peptide-corticotropin-releasing factor receptor interactions in caudal brainstem controls of solid food intake. Injections of combinations of vehicle, cocaine-and amphetamine-regulated transcript peptide (0.5 µg or 1 µg) or α-helical CRF9-41 were given into the fourth cerebral ventricle of rats. Nocturnal solid food intake was recorded over 22 h. RESULTS: Pre-treatment with α-helical CRF9-41 into the fourth ventricle significantly increased the responsivity to cocaine-and amphetamine-regulated transcript peptide on hypophagia. In a separate control experiment, α-helical CRF9-41 pre-treatment blocked CRF-induced food intake inhibition indicative of its antagonistic effectiveness. CONCLUSIONS: We conclude that an endogenous Corticotropin-releasing factor agonist may modulate suppression of food intake caused by cocaine-and amphetamine-regulated transcript peptide at a dorsal hindbrain level in the absence of stress. A potential caudal brainstem mechanism whereby cocaine-and amphetamine-regulated transcript peptide effects on food intake is attenuated via corticotropin-releasing factor receptor activity causing tonic inhibition, is suggested.


Assuntos
Fármacos do Sistema Nervoso Central/farmacologia , Hormônio Liberador da Corticotropina/antagonistas & inibidores , Ingestão de Alimentos/efeitos dos fármacos , Proteínas do Tecido Nervoso/farmacologia , Fragmentos de Peptídeos/farmacologia , Rombencéfalo/efeitos dos fármacos , Animais , Hormônio Liberador da Corticotropina/metabolismo , Hormônio Liberador da Corticotropina/farmacologia , Ingestão de Alimentos/fisiologia , Quarto Ventrículo , Injeções Intraventriculares , Masculino , Proteínas do Tecido Nervoso/metabolismo , Fragmentos de Peptídeos/metabolismo , Ratos Sprague-Dawley , Rombencéfalo/metabolismo
12.
J Pept Sci ; 25(4): e3153, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30734396

RESUMO

The aim of the study was to determine the in vitro immunomodulatory, cytotoxic, and insulin-releasing activities of seven phylloseptin-TR peptides and plasticin-TR, first isolated from the frog Phyllomedusa trinitatis. The most cationic peptides, phylloseptin-1.1TR and phylloseptin-3.1TR, showed greatest cytotoxic potency against A549, MDA-MB231, and HT-29 human tumor-derived cells and against mouse erythrocytes. Phylloseptin-4TR was the most hydrophobic and the most effective peptide at inhibiting production of the proinflammatory cytokines TNF-α and IL-1ß by mouse peritoneal cells but was without effect on production of the antiinflammatory cytokine IL-10. Phylloseptin-2.1TR and phylloseptin-3.3TR were the most effective at stimulating the production of IL-10. The noncytotoxic peptide, plasticin-TR, inhibited production of TNF-α and IL-1ß but was without effect on IL-10 production. The results of CD spectroscopy suggest that the different properties of plasticin-TR compared with the immunostimulatory activities of the previously characterized plasticin-L1 from Leptodactylus laticeps may arise from greater ability of plasticin-TR to oligomerize and adopt a stable helical conformation in a membrane-mimetic environment. All peptides stimulated release of insulin from BRIN-BD11 rat clonal ß cells with phylloseptin-3.2TR being the most potent and effective and phylloseptin-2.1TR the least effective suggesting that insulinotropic potency correlates inversely with helicity. The study has provided insight into structure-activity relationships among the phylloseptins. The combination of immunomodulatory and insulinotropic activities together with low cytotoxicity suggests that phylloseptin-3.3TR and plasticin-TR may represent templates for the development of agents for use in antiinflammatory and type 2 diabetes therapies.


Assuntos
Anti-Inflamatórios/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Anuros , Citotoxinas/farmacologia , Proteínas do Olho/farmacologia , Imunomodulação/efeitos dos fármacos , Insulina/metabolismo , Proteínas do Tecido Nervoso/farmacologia , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/imunologia , Anti-Inflamatórios/isolamento & purificação , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/imunologia , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Proteínas do Olho/química , Proteínas do Olho/imunologia , Proteínas do Olho/isolamento & purificação , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/imunologia , Proteínas do Tecido Nervoso/isolamento & purificação , Ratos
13.
Cancer Res ; 79(6): 1165-1177, 2019 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-30674535

RESUMO

The apoptosis repressor with caspase recruitment domain (ARC) protein is a strong independent adverse prognostic marker in acute myeloid leukemia (AML). We previously reported that ARC regulates leukemia-microenvironment interactions through the NFκB/IL1ß signaling network. Malignant cells have been reported to release IL1ß, which induces PGE2 synthesis in mesenchymal stromal cells (MSC), in turn activating ß-catenin signaling and inducing the cancer stem cell phenotype. Although Cox-2 and its enzymatic product PGE2 play major roles in inflammation and cancer, the regulation and role of PGE2 in AML are largely unknown. Here, we report that AML-MSC cocultures greatly increase Cox-2 expression in MSC and PGE2 production in an ARC/IL1ß-dependent manner. PGE2 induced the expression of ß-catenin, which regulated ARC and augmented chemoresistance in AML cells; inhibition of ß-catenin decreased ARC and sensitized AML cells to chemotherapy. NOD/SCIDIL2RγNull-3/GM/SF mice transplanted with ARC-knockdown AML cells had significantly lower leukemia burden, lower serum levels of IL1ß/PGE2, and lower tissue human ARC and ß-catenin levels, prolonged survival, and increased sensitivity to chemotherapy than controls. Collectively, we present a new mechanism of action of antiapoptotic ARC by which ARC regulates PGE2 production in the tumor microenvironment and microenvironment-mediated chemoresistance in AML.Significance: The antiapoptotic protein ARC promotes AML aggressiveness by enabling detrimental cross-talk with bone marrow mesenchymal stromal cells.


Assuntos
Ciclo-Oxigenase 2/metabolismo , Proteínas do Citoesqueleto/farmacologia , Dinoprostona/farmacologia , Resistencia a Medicamentos Antineoplásicos , Interleucina-1beta/metabolismo , Leucemia Mieloide Aguda/patologia , Proteínas do Tecido Nervoso/farmacologia , Microambiente Tumoral , beta Catenina/metabolismo , Animais , Apoptose , Biomarcadores Tumorais/metabolismo , Proliferação de Células , Ciclo-Oxigenase 2/genética , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-1beta/genética , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/metabolismo , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/patologia , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Ocitócicos/farmacologia , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto , beta Catenina/genética
14.
PLoS One ; 14(1): e0207190, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30673694

RESUMO

PURPOSE: To analyze the potential neuro-protective and neuro-regenerative effects of Collapsin-response-mediator-protein-5 (CRMP-5) on retinal ganglion cells (RGCs) using in vitro and in vivo animal models of glaucoma. METHODS: Elevated intraocular pressure (IOP) was induced in adult female Sprague-Dawley (SD) rats by cauterization of three episcleral veins. Changes in CRMP-5 expression within the retinal proteome were analyzed via label-free mass spectrometry. In vitro, retinal explants were cultured under elevated pressure (60 mmHg) within a high-pressure incubation chamber with and without addition of different concentrations of CRMP-5 (4 µg/l, 200 µg/l and 400 µg/l). In addition, retinal explants were cultured under regenerative conditions with and without application of 200 µg/l CRMP-5 after performing an optic nerve crush (ONC). Thirdly, an antibody against Protein Kinase B (PKB) was added to examine the possible effects of CRMP-5. RGC count was performed. Number and length of the axons were determined and compared. To undermine a signal-transduction pathway via CRMP-5 and PKB microarray and immunohistochemistry were performed. RESULTS: CRMP-5 was downregulated threefold in animals showing chronically elevated IOP. The addition of CRMP-5 to retinal culture significantly increased RGC numbers under pressure in a dose-dependent manner and increased and elongated outgrowing axons in retinal explants significantly which could be blocked by PKB. Especially the number of neurites longer than 400 µm significantly increased after application of CRMP-5. CRMP-5 as well as PKB were detected higher in the experimental than in the control group. CONCLUSION: CRMP-5 seems to play an important role in an animal model of glaucoma. Addition of CRMP-5 exerts neuro-protective and neuro-regenerative effects in vitro. This effect could be mediated via activation of PKB affecting intra-cellular apoptosis pathways.


Assuntos
Glaucoma/patologia , Glaucoma/fisiopatologia , Modelos Biológicos , Regeneração Nervosa/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Células Ganglionares da Retina/patologia , Animais , Feminino , Glaucoma/metabolismo , Proteínas do Tecido Nervoso/farmacologia , Crescimento Neuronal/efeitos dos fármacos , Neuroproteção/efeitos dos fármacos , Proteoma/metabolismo , Proteômica , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos Sprague-Dawley , Células Ganglionares da Retina/efeitos dos fármacos
15.
Addict Biol ; 24(1): 51-64, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-29193459

RESUMO

Although chronic nicotine administration does not affect memory, its withdrawal causes massive cognitive deficits. The underlying mechanisms, however, have not been understood. We test the role of cocaine- and amphetamine-regulated transcript peptide (CART), a neuropeptide known for its procognitive properties, in this process. The mice on chronic nicotine treatment/withdrawal were subjected to novel object recognition task. The capability of the animal to discriminate between the novel and familiar objects was tested and represented as discrimination index (DI); reduction in the index suggested amnesia. Nicotine for 49 days had no effect on DI, but 8-hour withdrawal caused a significant reduction, followed by full recovery at 24-hour withdrawal timepoint. Bilateral CART infusion in dorsal hippocampus rescued deficits in DI at 8-hours, whereas CART-antibody infusion into the dorsal hippocampus attenuated the recovery at 24-hours. Commensurate changes were observed in the CART as well as CART mRNA profiles in the hippocampus. CART mRNA expression and the peptide immunoreactivity did not change significantly following chronic nicotine treatment. However, there was a significant reduction at 8-hour withdrawal, followed by a drastic increase in CART immunoreactivity as well as CART mRNA at 24-hour withdrawal, compared with 8-hour withdrawal. Distinct α7-nicotinic receptor immunoreactivity was detected on the hippocampal CART neurons, suggesting cholinergic inputs. An increase in the synaptophysin immunoreactive elements around CART cells in the dentate gyrus, cornu ammonis 3 and subiculum at 24-hour post-withdrawal timepoint suggested neuronal plasticity. CART circuit dynamics in the hippocampus seems to modulate short-term memory associated with nicotine withdrawal.


Assuntos
Proteínas do Tecido Nervoso/farmacologia , Nicotina/efeitos adversos , Agonistas Nicotínicos/efeitos adversos , Reconhecimento Psicológico/efeitos dos fármacos , Síndrome de Abstinência a Substâncias/psicologia , Animais , Região CA3 Hipocampal/citologia , Região CA3 Hipocampal/efeitos dos fármacos , Região CA3 Hipocampal/metabolismo , Giro Denteado/citologia , Giro Denteado/efeitos dos fármacos , Giro Denteado/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Camundongos , Proteínas do Tecido Nervoso/antagonistas & inibidores , Proteínas do Tecido Nervoso/genética , Plasticidade Neuronal , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Síndrome de Abstinência a Substâncias/etiologia , Síndrome de Abstinência a Substâncias/genética , Sinaptofisina/efeitos dos fármacos , Sinaptofisina/metabolismo
16.
Metab Brain Dis ; 34(1): 79-85, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30269302

RESUMO

Nesfatin-1 is a novel peptide with anorexigenic and anti-hyperglycemic properties. According to previous studies, this multi-functional peptide protects dopaminergic cells against neurotoxicity via anti-apoptotic effects. In addition, Nesfatin-1 protects myocardial tissue after myocardial infarction via anti-inflammatory and anti-apoptotic mechanisms. In this study, we investigated the neuroprotective effects of nesfatin-1 against cerebral ischemia reperfusion injury in the CA1 area of hippocampus in rats. 56 male Wistar rats (240-270 g) were randomly selected and allocated into four groups: (1) sham, (2) nesfatin-1, (3) ischemia/reperfusion, (4) ischemia/reperfusion+nesfatin-1. Cerebral ischemia induced by the occlusion of the common carotid arteries for 20 min was followed by reperfusion. Saline as a vehicle and nesfatin-1 (20 µg/kg) were injected intraperitoneally (IP) at the start of cerebral reperfusion. Apoptotic and necrotic cell death was detected by TUNEL and Nissl staining. Malondialdehyde (MDA) and antioxidant enzymes (GSH and SOD) levels were measured by the ELISA method. The results showed that cerebral ischemia increased the apoptotic and necrotic cell death in the CA1 area of hippocampus, while, treatment with nesfatin-1significantly reduced apoptotic and necrotic cell death. Moreover, the MDA levels of the hippocampus in ischemic rats were higher, whereas in nesfatin-1-treated rats the MDA levels were decreased. Furthermore, the SOD and GSH levels in the ischemic rats were decreased, whilst in ischemic rats treated with nesfatin-1, the SOD and GSH levels were increased. This study for the first time found that nesfatin-1 treatment improves CA1 hippocampus injuries after cerebral ischemia through preventing neuronal cell death and enhancement of antioxidant defenses.


Assuntos
Antioxidantes/uso terapêutico , Proteínas de Ligação ao Cálcio/uso terapêutico , Morte Celular/efeitos dos fármacos , Proteínas de Ligação a DNA/uso terapêutico , Hipocampo/efeitos dos fármacos , Proteínas do Tecido Nervoso/uso terapêutico , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/uso terapêutico , Traumatismo por Reperfusão/prevenção & controle , Animais , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Proteínas de Ligação ao Cálcio/farmacologia , Caspase 3/metabolismo , Proteínas de Ligação a DNA/farmacologia , Modelos Animais de Doenças , Hipocampo/metabolismo , Masculino , Malondialdeído/metabolismo , Proteínas do Tecido Nervoso/farmacologia , Neurônios/metabolismo , Fármacos Neuroprotetores/farmacologia , Nucleobindinas , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Superóxido Dismutase/metabolismo
17.
Gen Comp Endocrinol ; 271: 49-60, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30391240

RESUMO

Recent studies have revealed nesfatin-1 as a hypothalamic neuropeptide, regulating food intake, energy expenditure and reproduction primarily by acting on the hypothalamic-pituitary-gonadal axis. Nesfatin-1 is also localized in several peripheral tissues including testes. However, functional significance of nesfatin-1 in testicular activities is not yet well documented in mammals. Therefore, this study was aimed to elucidate the direct effects of nesfatin-1 on testicular markers for steroid productions, spermatogenesis, metabolic changes and oxidative stress. The results revealed the expression of both protein and mRNA of nesfatin-1 in the testes of adult mice. The testes treated in vitro with nesfatin-1 showed significant increase in testosterone production, which correlated significantly with increased expression of steroidogenic markers and insulin receptor proteins in the testes. Furthermore, the in vitro treatment with nesfatin-1 showed stimulatory effects on spermatogenesis by promoting cell proliferation (PCNA) and survival (Bcl2), while inhibiting apoptosis (caspase-3) in the testes. The nesfatin-1 treatment in vitro further increased the expression of insulin receptor and GLUT8 proteins, in parallel with increase in the intra-testicular transport of glucose and production of lactate. This nesfatin-1 induced enhanced transport of energy substrate (glucose and lactate) may be responsible for promoting spermatogenesis and steroidogenesis. Nesfatin-1 significantly reduced oxidative stress and nitric oxide, which may also be responsible for stimulatory effects on testicular activities. The present finding suggests that nesfatin-1 acts via paracrine manner to increase sperm count and fertility, thus promoting the testicular function.


Assuntos
Envelhecimento/metabolismo , Proteínas de Ligação ao Cálcio/farmacologia , Proteínas de Ligação a DNA/farmacologia , Proteínas do Tecido Nervoso/farmacologia , Neuropeptídeos/farmacologia , Espermatogênese , Esteroides/biossíntese , Testículo/metabolismo , Animais , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Estradiol/metabolismo , Masculino , Camundongos , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Nucleobindinas , Estresse Oxidativo/efeitos dos fármacos , Antígeno Nuclear de Célula em Proliferação/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Testículo/efeitos dos fármacos , Testosterona/metabolismo
18.
Acta Neuropathol Commun ; 6(1): 121, 2018 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-30409172

RESUMO

Pathologic, biochemical and genetic evidence indicates that accumulation and aggregation of amyloid ß-proteins (Aß) is a critical factor in the pathogenesis of Alzheimer's disease (AD). Several therapeutic interventions attempting to lower Aß have failed to ameliorate cognitive decline in patients with clinical AD significantly, but most such approaches target only one or two facets of Aß production/clearance/toxicity and do not consider the heterogeneity of human Aß species. As synaptic dysfunction may be among the earliest deficits in AD, we used hippocampal long-term potentiation (LTP) as a sensitive indicator of the early neurotoxic effects of Aß species. Here we confirmed prior findings that soluble Aß oligomers, much more than fibrillar amyloid plaque cores or Aß monomers, disrupt synaptic function. Interestingly, not all (84%) human AD brain extracts are able to inhibit LTP and the degree of LTP impairment by AD brain extracts does not correlate with Aß levels detected by standard ELISAs. Bioactive AD brain extracts also induce neurotoxicity in iPSC-derived human neurons. Shorter forms of Aß (including Aß1-37, Aß1-38, Aß1-39), pre-Aß APP fragments (- 30 to - 1) and N-terminally extended Aßs (- 30 to + 40) each showed much less synaptotoxicity than longer Aßs (Aß1-42 - Aß1-46). We found that antibodies which target the N-terminus, not the C-terminus, efficiently rescued Aß oligomer-impaired LTP and oligomer-facilitated LTD. Our data suggest that preventing soluble Aß oligomer formation and targeting their N-terminal residues with antibodies could be an attractive combined therapeutic approach.


Assuntos
Doença de Alzheimer/patologia , Hipocampo/patologia , Sinapses/patologia , Sinapses/fisiologia , Doença de Alzheimer/líquido cefalorraquidiano , Doença de Alzheimer/genética , Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/imunologia , Peptídeos beta-Amiloides/metabolismo , Peptídeos beta-Amiloides/farmacologia , Precursor de Proteína beta-Amiloide/genética , Animais , Anticorpos/farmacologia , Fatores de Transcrição Hélice-Alça-Hélice Básicos/farmacologia , Modelos Animais de Doenças , Feminino , Humanos , Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/metabolismo , Potenciação de Longa Duração/efeitos dos fármacos , Potenciação de Longa Duração/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteínas do Tecido Nervoso/metabolismo , Proteínas do Tecido Nervoso/farmacologia , Inibição Neural/efeitos dos fármacos , Inibição Neural/genética , Fragmentos de Peptídeos/farmacologia , Tirosina/análogos & derivados , Tirosina/farmacologia
19.
Invest Ophthalmol Vis Sci ; 59(12): 5277-5284, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30383199

RESUMO

Purpose: Corneal vascularization significantly increases the risk for graft rejection after keratoplasty. Semaphorin 3F (Sema3F) is a known modulator of physiologic avascularity in the outer retina. The aim of this study was to investigate whether Sema3F is involved in maintaining corneal avascularity and can reduce the risk for corneal graft rejection. Methods: Corneal Sema3F expression was investigated using immunohistochemistry and qPCR in human and murine tissue. Pathologic invasion of blood and lymph vessels into corneal tissue was analyzed in the murine corneal suture and high-risk keratoplasty model. The anti-lymphangiogenic effects of Sema3F were further investigated using an in vitro spheroidal sprouting model with supernatant from isolated primary human corneal epithelial cells (hCECs). Results: Sema3F is constitutively expressed in human and murine corneal epithelium. In the corneal suture model, lymphangiogenesis was significantly suppressed by topical Sema3F treatment (P = 0.0003). In the murine high-risk keratoplasty model, pretreatment by topical Sema3F in the inflammation phase significantly promoted subsequent graft survival (P = 0.0006). In this model, both lymph- and blood angiogenesis were reduced (P < 0.05). In vitro, hCEC supernatant had a direct anti-lymphangiogenic effect on human lymphatic endothelial cells (P < 0.01). This effect was completely abolished by addition of anti-Sema3F antibodies. Conclusions: Sema3F is a novel mediator of corneal avascularity with potent anti-lymphangiogenic properties. Topical treatment with Sema3F eye drops may help to limit corneal vascularization and improve outcomes in high-risk keratoplasty patients.


Assuntos
Neovascularização da Córnea/prevenção & controle , Sobrevivência de Enxerto/efeitos dos fármacos , Ceratoplastia Penetrante , Linfangiogênese/efeitos dos fármacos , Proteínas de Membrana/farmacologia , Proteínas do Tecido Nervoso/farmacologia , Animais , Proliferação de Células , Modelos Animais de Doenças , Epitélio Anterior/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Vasos Linfáticos/metabolismo , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Proteínas do Tecido Nervoso/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
20.
Biomed Mater Eng ; 29(6): 757-771, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30282332

RESUMO

INTRODUCTION: Exogenous Nel-like molecule type 1 (NELL1) represents a potentially attractive clinical treatment option in the orthodontic and other settings because of its osteoinductive and vasculogenic properties. AIMS: To explore effects of NELL1on corticotomy-assisted tooth movement and osteogenesis in a rat model. METHODS: Thirty Sprague-Dawley rats were divided into 6 groups: Control, Sham, Tooth movement only, Vehicle, NELL1-LD (low-dose NELL), NELL1-HD (high-dose NELL). Human recombinant NELL1 protein was applied locally (Groups NELL1-LD and NELL1-HD) into buccal mucosa region of left first upper molar. Then the distance and velocity of tooth movement was measured, animals at 6 weeks after surgery were sacrificed, and was followed by computed tomography and histochemical staining. RESULTS: Both NELL1 groups had higher bone mineral density, greater tooth movement distance and velocity in comparison to the Vehicle group. Proximally and distally, periodontal ligament width was significantly increased in the NELL1-LD and NELL1-HD groups. Decortication enhances remodeling, however, rapid bone formation by high-dose NELL1 may affect bone absorption. CONCLUSION: Appropriate dose of NELL1 can be administrated to reduce the total time for tooth movement, and may shorten the treatment time in select populations.


Assuntos
Ligamentos/fisiologia , Proteínas do Tecido Nervoso/farmacologia , Osteogênese/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Técnicas de Movimentação Dentária/métodos , Animais , Densidade Óssea , Humanos , Masculino , Maxila , Dente Molar , Ortodontia , Ratos , Ratos Sprague-Dawley , Microtomografia por Raio-X
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