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1.
Nat Commun ; 12(1): 2086, 2021 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-33828102

RESUMO

Histamine receptors play important roles in various pathophysiological conditions and are effective targets for anti-allergy treatment, however the mechanism of receptor activation remain elusive. Here, we present the cryo-electron microscopy (cryo-EM) structure of the human H1R in complex with a Gq protein in an active conformation via a NanoBiT tethering strategy. The structure reveals that histamine activates receptor via interacting with the key residues of both transmembrane domain 3 (TM3) and TM6 to squash the binding pocket on the extracellular side and to open the cavity on the intracellular side for Gq engagement in a model of "squash to activate and expand to deactivate". The structure also reveals features for Gq coupling, including the interaction between intracellular loop 2 (ICL2) and the αN-ß junction of Gq/11 protein. The detailed analysis of our structure will provide a framework for understanding G-protein coupling selectivity and clues for designing novel antihistamines.


Assuntos
Microscopia Crioeletrônica/métodos , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/química , Histamina/química , Receptores Histamínicos/química , Sítios de Ligação , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/metabolismo , Proteínas de Ligação ao GTP , Histamina/metabolismo , Humanos , Ligantes , Ligação Proteica , Domínios Proteicos , Receptores Histamínicos/metabolismo , Receptores Histamínicos H1/metabolismo
2.
Biochem Pharmacol ; 184: 114401, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33387483

RESUMO

BACKGROUND: Allergic asthma is a common inflammatory lung disease associated with complex pathogenesis. Mast cell (MC) is one of the key drivers of allergic asthma, Mas-related G protein-coupled receptor X2 (MRGPRX2) on the MC could mediate MC activation and trigger a pseudo-allergic reaction. Imperatorin (IMP), the main active compound of Radix Angelicae Dahuricae, has been reported to exert various pharmacological effects. In this study, we focused on the therapeutical mechanism of IMP on MRGPRX2-induced pseudo-allergy and allergic asthma. METHODS: We examined the effect of IMP on MRGPRX2 related mast cell activation in mouse peritoneal MC (MPMC), Human Laboratory of Allergic Disease 2 MCs (LAD2 cells) and Mrgprx2-expressing HEK293 cells. Molecular docking and Surface plasmon resonance (SPR) were taken to reveal the binding character between IMP and MRPGRX2. MRGPRX2 downstream proteins were also detected by western blotting. IgE-independent responses was evaluated by using passive cutaneous anaphylaxis (PCA) and active systemic anaphylaxis (ASA) models. The therapeutic effect of IMP on asthma was evaluated by a lung inflammation mouse model which was induced by ovalbumin (OVA). RESULTS: IMP was found to reduce substance P (SP) induced calcium flux and suppressed degranulation of MC. SP can promote the phosphorylation of ERK and CamKII, which regulates the synthesis of inflammatory factors such as MIP-2 and TNF-α in MC. In vivo assays revealed that IMP can mitigate SP-induced mouse PCA and ASA. IMP could also mitigate lung inflammation in an OVA induced mice model by inhibiting MC activation in the lung tissue. Furthermore, IMP binds well to MRGPRX2 protein. The binding constant (KD) is 4.48 ± 0.49 × 10-7 M. The data suggeste that IMP is a novel inhibitor of MRGPRX2 to treat allergic asthma.


Assuntos
Furocumarinas/farmacologia , Hipersensibilidade/tratamento farmacológico , Mastócitos/efeitos dos fármacos , Receptores Acoplados a Proteínas-G/antagonistas & inibidores , Animais , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Linhagem Celular , Furocumarinas/metabolismo , Histamina/metabolismo , Humanos , Hipersensibilidade/metabolismo , Hipersensibilidade/patologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Mastócitos/metabolismo , Mastócitos/patologia , Camundongos Endogâmicos C57BL , Anafilaxia Cutânea Passiva/efeitos dos fármacos , Pneumonia/tratamento farmacológico , Pneumonia/etiologia , Receptores Acoplados a Proteínas-G/química , Receptores Acoplados a Proteínas-G/metabolismo
3.
Yakugaku Zasshi ; 141(1): 93-110, 2021.
Artigo em Japonês | MEDLINE | ID: mdl-33390452

RESUMO

There has been little information about the role of histamine on the central nervous system (CNS), different from dopamine and serotonin. In the present study, therefore, the effects of histamine and related compounds on the CNS were studied using rats. Intracerebroventricular (i.c.v.) injection of histamine and 2-methylhistamine ameliorated memory deficit after long interrution of learning in active avoidance response. First generation H1-antagonists inhibited active avoidance response, whereas newly develpoed H1-antagonists showed little effect. α-Fluoromethylhistidine, an histidine decarboxylase inhibitor, also inhibited active avoidance response. In radial maze performance, almost the same findings were obtained. I.c.v. injection of histamine and H1-agonists inhibited amygdaloid kindled seizures. First generation H1-antagonists attenuated histamine-induced inhibition of amygdaloid kindled seizures. Both i.c.v. and intraperitoneal injections of H3-antagonist, thioperamide, resulted in a dose-related inhibition of amygdaloid kindled seizures. The effect of thioperamide was inhibited by an H3-agonists and H1-antagonists. Similar to nitrazepam, diphenhydramine and chlorpheniramine caused a shortening of sleep latency. On the other hand, no significant effects were observed with second generation H1-antagonists. These findings suggest that histamine plays an important role in learning and memory via H1-receptors, an inhibition of amygdaloid kindled seizures induced by histamine occurred through not only H1-receptors but also H3-receptors, and that classic H1-antagonists can be useful as a effective hypnotic for difficulty in falling asleep.


Assuntos
Sistema Nervoso Central/efeitos dos fármacos , Antagonistas dos Receptores Histamínicos H1/farmacologia , Histamina/farmacologia , Metilistaminas/farmacologia , Metilistidinas/farmacologia , Animais , Aprendizagem da Esquiva/efeitos dos fármacos , Histamina/administração & dosagem , Histamina/metabolismo , Histamina/fisiologia , Antagonistas dos Receptores Histamínicos H1/uso terapêutico , Humanos , Hipnóticos e Sedativos , Injeções Intraventriculares , Excitação Neurológica/efeitos dos fármacos , Transtornos da Memória/tratamento farmacológico , Metilistaminas/administração & dosagem , Metilistidinas/administração & dosagem , Camundongos , Ratos , Receptores Histamínicos H3/metabolismo , Receptores Histamínicos H3/fisiologia , Convulsões/tratamento farmacológico , Sono/efeitos dos fármacos , Distúrbios do Início e da Manutenção do Sono/tratamento farmacológico
4.
Food Chem ; 343: 128397, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33406569

RESUMO

Chocolate is an important source of free bioactive amines and amino acids which play important roles in human health. Considering the limited information on the bioaccessibility of these compounds from chocolate, the objective of this study was to characterize their profiles and bioaccessibility in 70% cocoa dark chocolate through in vitro simulation of oral, gastric and intestinal digestions. Seven amines were detected; polyamines were predominant before in vitro digestion, whereas tyramine, cadaverine and spermidine after digestion. All amines showed high bioaccessibility with slight influence of digestive enzymes. Amines increased after gastrointestinal digestion: tyramine (13-fold), tryptamine (9-fold), others (2.4-4.2-fold) and histamine appeared. All amino acids, GABA and ammonia were detected in chocolate, and their contents increased after in vitro digestion due to digestive enzymes (4.6, 2.8 and 2.1, respectively). Dark chocolate protein is a good source of tryptophan, phenylalanine + tyrosine, isoleucine, histidine, but limiting for lysine, leucine, and threonine.


Assuntos
Aminas/metabolismo , Aminoácidos/metabolismo , Cacau/metabolismo , Chocolate/análise , Aminas/análise , Aminoácidos/análise , Amônia/metabolismo , Cacau/química , Cromatografia Líquida de Alta Pressão , Análise por Conglomerados , Histamina/análise , Histamina/metabolismo , Humanos , Valor Nutritivo , Poliaminas/análise , Poliaminas/metabolismo , Análise de Componente Principal , Ácido gama-Aminobutírico/análise
5.
BMJ Case Rep ; 13(12)2020 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-33310825

RESUMO

Histamine is involved in various physiological functions like sleep-wake cycle and stress regulation. The histamine N-methyltransferase (HNMT) enzyme is the only pathway for termination of histamine neurotransmission in the central nervous system. Experiments with HNMT knockout mice generated aggressive behaviours and dysregulation of sleep-wake cycles. Recently, seven members of two unrelated consanguineous families have been reported in whom two different missense HNMT mutations were identified. All showed severe intellectual disability, delayed speech development and mild regression from the age of 5 years without, however, any dysmorphisms or congenital abnormality. A diagnosis of mental retardation, autosomal recessive 51 was made. Here, we describe a severely mentally retarded adolescent male born from second cousins with a homozygous mutation in HNMT. His phenotypic profile comprised aggression, delayed speech, autism, sleep disturbances and gastro-intestinal problems. At early age, regression occurred. Treatment with hydroxyzine combined with a histamine-restricted diet resulted in significant general improvement.


Assuntos
Histamina N-Metiltransferase/genética , Homozigoto , Deficiência Intelectual/genética , Mutação , Agressão/fisiologia , Encéfalo/metabolismo , Histamina/metabolismo , Histamina N-Metiltransferase/metabolismo , Humanos , Hidroxizina/uso terapêutico , Deficiência Intelectual/dietoterapia , Deficiência Intelectual/tratamento farmacológico , Deficiência Intelectual/metabolismo , Masculino , Sono/fisiologia , Resultado do Tratamento , Adulto Jovem
6.
PLoS One ; 15(10): e0239672, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33002062

RESUMO

Human Fortilin, an antiapoptotic protein, has also been implicated in several diseases; however, several potential uses of fortilin have also been proposed. Bearing the implications of fortilin in mind, fortilin analog, which has no complication with diseases, is required. Since a recombinant full-length fortilin from Fenneropenaeus merguiensis (rFm-Fortilin (FL)) reported only 44% (3e-27) homologous to human fortilin, therefore the biological activities of the Fm-Fortilin (FL) and its fragments (F2, F12, and F23) were investigated for potential use against HEMA toxicity from filling cement to pulp cell. The rFm-Fortilin FL, F2, 12, and F23 were expressed and assayed for proliferation activity. The rFm-Fortilin (FL) showed proliferation activity on human dental pulp cells (HDPCs) and protected the cells from 2-hydroxy-ethyl methacrylate (HEMA) at 1-20 ng/ml. In contrast, none of the rFm-Fortilin fragments promoted HDPC growth that may be due to a lack of three conserved amino acid residues together for binding with the surface of Rab GTPase for proliferative activity. In addition, rFm-Fortilin (FL) activated mineralization and trend to suppressed production of proinflammatory cytokines, including histamine (at 10 ng/ml) and TNF-α (at 100 ng/ml). Besides, the rFm-Fortilin (FL) did not mutate the Chinese hamster ovary (CHO) cell. Therefore, the rFm-Fortilin (FL) has the potential use as a supplementary medical material to promote cell proliferation in patients suffering severe tooth decay and other conditions.


Assuntos
Proteínas de Artrópodes/farmacologia , Penaeidae/química , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/isolamento & purificação , Células Cultivadas , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Histamina/metabolismo , Metacrilatos/toxicidade , Proteínas Recombinantes , Alinhamento de Sequência , Fator de Necrose Tumoral alfa/metabolismo
7.
Extremophiles ; 24(5): 787-796, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32743715

RESUMO

Salted and ripened fish foods are susceptible to cause histamine poisoning. The present study focuses on microbial histamine degradation from high salted fermented fishery products to deepen our understanding about this new and growing field of research. As a result of this first study related to salted-ripened anchovies (Engraulis anchoita), fifty seven moderate and extreme halophilic microbial isolates from salt and salted-ripened anchovy processes were characterized in terms of their phenotype and histamine-degrading capacity. Only 7%-4 isolates-were able to degrade histamine. None of the histamine-degrading isolates presented proteolytic and/or lipolytic activity. One of them designated A18 was chemotactic toward histamine, an interesting property not previously reported for that chemoattractant. However, the S18 and A18 isolates, genotypically identified as Halobacterium sp. and Halomonas sp. respectively, produced indole and/or H2S, both undesirable characteristics associated to off-flavors occurrence. On the other hand, A28 and S20, identified as Halovibrio sp. and Halobacterium sp. respectively, presented desirable properties, such as cytochrome oxidase and catalase activity, and non-production of H2S and indole. These strains also showed characteristics previously reported as dominant in the ripened stage. The results are promising, and A28 and S20 may have the desirable features to improve the anchovy salting-ripening process.


Assuntos
Halobacteriales , Halomonas , Histamina , Animais , Aquicultura , Peixes , Histamina/metabolismo , RNA Ribossômico 16S , Cloreto de Sódio
8.
Proc Natl Acad Sci U S A ; 117(30): 18068-18078, 2020 07 28.
Artigo em Inglês | MEDLINE | ID: mdl-32661165

RESUMO

Mast cells and basophils are main drivers of allergic reactions and anaphylaxis, for which prevalence is rapidly increasing. Activation of these cells leads to a tightly controlled release of inflammatory mediators stored in secretory granules. The release of these granules is dependent on intracellular calcium (Ca2+) signals. Ca2+ release from endolysosomal compartments is mediated via intracellular cation channels, such as two-pore channel (TPC) proteins. Here, we uncover a mechanism for how TPC1 regulates Ca2+ homeostasis and exocytosis in mast cells in vivo and ex vivo. Notably, in vivo TPC1 deficiency in mice leads to enhanced passive systemic anaphylaxis, reflected by increased drop in body temperature, most likely due to accelerated histamine-induced vasodilation. Ex vivo, mast cell-mediated histamine release and degranulation was augmented upon TPC1 inhibition, although mast cell numbers and size were diminished. Our results indicate an essential role of TPC1 in endolysosomal Ca2+ uptake and filling of endoplasmic reticulum Ca2+ stores, thereby regulating exocytosis in mast cells. Thus, pharmacological modulation of TPC1 might blaze a trail to develop new drugs against mast cell-related diseases, including allergic hypersensitivity.


Assuntos
Anafilaxia/etiologia , Anafilaxia/metabolismo , Canais de Cálcio/deficiência , Suscetibilidade a Doenças , Mastócitos/imunologia , Mastócitos/metabolismo , Biomarcadores , Sinalização do Cálcio , Degranulação Celular , Citocinas/metabolismo , Predisposição Genética para Doença , Histamina/metabolismo , Imunoglobulina E/imunologia , Mediadores da Inflamação/metabolismo
9.
Food Chem ; 331: 127347, 2020 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-32574945

RESUMO

Biogenic amines (BAs) are a class of bioactive organics produced during the fermentation of soy sauce. A high concentration of BAs may bring about serious physiological and toxicological effects on the human body. In this study, we reported an optimized process to produce soy sauce with lower BA concentration and found the contents of putrescine, cadaverine and histamine increased with the increase of fermentation temperature but decreased with the increase of NaCl concentration. The final content of total BAs with improved fermentation was 105.56 ± 0.13 mg/L, which was reduced by 89.11% compared to traditional brewing. Besides, the pilot production test was performed to verify the optimized conditions and physicochemical indexes were measured to better understand the change principle of the chemical compounds. Taken together, we present an effective process to inhibit the formation of BAs while ensuring that characteristic nutrients are not lost.


Assuntos
Aminas Biogênicas/biossíntese , Alimentos de Soja , Aminas Biogênicas/análise , Cadaverina/análise , Cadaverina/metabolismo , Fermentação , Indústria de Processamento de Alimentos/métodos , Histamina/análise , Histamina/metabolismo , Putrescina/análise , Putrescina/metabolismo , Cloreto de Sódio , Alimentos de Soja/análise , Temperatura
10.
Food Chem ; 331: 127355, 2020 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-32593042

RESUMO

Ara h1 is a major allergen from peanut. We investigated the effect of covalent conjugation of Ara h1 and dietary polyphenols on allergenicity and functional properties of Ara h1. Enzyme-linked immunosorbent assay revealed that the covalent conjugation of dietary polyphenols significantly reduced the IgE binding capacity of Ara h1. Covalent binding of dietary polyphenols with Ara h1 reduced histamine release by 40% in basophils. The decreased IgE binding capacity of Ara h1 could be ascribed to changes in protein conformation. The IgE epitope of Ara h1 might be blocked by polyphenols at the binding site. Analysis of pepsin digestion of Ara h1-polyphenol conjugates indicated that the covalent binding increased pepsin digestibility and reduced IgE binding capacity. Furthermore, covalent conjugation of Ara h1 with polyphenols decreased denaturation temperature and increased antioxidant activity. Ara h1 conjugated with polyphenols may be a promising approach for reducing the allergenicity of Ara h1.


Assuntos
Antígenos de Plantas/química , Antígenos de Plantas/imunologia , Catequina/análogos & derivados , Ácido Clorogênico/química , Proteínas de Membrana/química , Proteínas de Membrana/imunologia , Hipersensibilidade a Amendoim/imunologia , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Antígenos de Plantas/farmacologia , Antioxidantes/química , Arachis/química , Basófilos/efeitos dos fármacos , Basófilos/imunologia , Basófilos/metabolismo , Catequina/química , Catequina/imunologia , Catequina/metabolismo , Epitopos/metabolismo , Histamina/metabolismo , Humanos , Imunoglobulina E/metabolismo , Proteínas de Membrana/farmacologia , Proteínas de Plantas/farmacologia , Conformação Proteica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectroscopia de Infravermelho com Transformada de Fourier
12.
Genes Cells ; 25(7): 443-449, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32394600

RESUMO

Histamine is a bioactive monoamine that is synthesized by the enzymatic activity of histidine decarboxylase (HDC) in basophils, mast cells, gastric enterochromaffin-like (ECL) cells and histaminergic neuronal cells. Upon a series of cellular stimuli, these cells release stored histamine, which elicits allergies, inflammation, and gastric acid secretion and regulates neuronal activity. Recent studies have shown that certain other types of myeloid lineage cells also produce histamine with HDC induction under various pathogenic stimuli. Histamine has been shown to play a series of pathophysiological roles by modulating immune and inflammatory responses in a number of disease conditions, whereas the mechanistic aspects underlying induced HDC expression remain elusive. In the present review, we summarize the current understanding of the regulatory mechanism of Hdc gene expression and the roles played by histamine in physiological contexts as well as pathogenic processes. We also introduce a newly developed histaminergic cell-monitoring transgenic mouse line (Hdc-BAC-GFP) that serves as a valuable experimental tool to identify the source of histamine and dissect upstream regulatory signals.


Assuntos
Histamina/metabolismo , Histidina Descarboxilase/metabolismo , Receptores Histamínicos/metabolismo , Sepse/imunologia , Animais , Cromossomos Artificiais Bacterianos , Regulação Enzimológica da Expressão Gênica/imunologia , Histamina/fisiologia , Histidina Descarboxilase/genética , Histonas/metabolismo , Metilação , Camundongos , Camundongos Transgênicos , Células Mieloides/metabolismo , Sepse/metabolismo
13.
Food Chem ; 321: 126723, 2020 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-32276143

RESUMO

A precise and unambiguous quantitative strategy for six biogenic amines in dry fermented mutton sausage with liquid chromatography tandem quadrupole Orbitrap mass spectrometry was developed to investigate the inhibitory effects of star anise, amomum tsao-ko, clove, cassia, fennel, bay leaf, and nutmeg on the accumulation of biogenic amines. Compared the data-dependent MS2 fragmentation (dd-MS2), variable data-independent acquisition (vDIA), and multiplex data-independent acquisition (mDIA), and the mDIA acquisition mode was selected for further analysis. The LODs and LOQs were 0.9-1.5 µg kg-1 and 2.9-5.0 µg kg-1. The maximum inhibition of spice on tryptamine, putrescine, spermidine, 2-phenylethylamine, tyrosamine, and histamine were 21.8%, 19.3%, 27.5%, 24.6%, 18.7% and 24.4%, which revealed that the accumulation of BAs was significantly inhibited by spice extracts, while cassia and fennel extracts showed the best effect. Thus, the established method can provide a better reference for dry fermented mutton sausage quality, safety and trade analysis.


Assuntos
Aminas Biogênicas/análise , Alimentos e Bebidas Fermentados/análise , Produtos da Carne , Especiarias , Aminas Biogênicas/metabolismo , Cassia/química , Cromatografia Líquida de Alta Pressão , Fermentação , Foeniculum , Cromatografia Gasosa-Espectrometria de Massas , Histamina/metabolismo , Produtos da Carne/análise , Extratos Vegetais/análise , Extratos Vegetais/química , Carne Vermelha/análise , Especiarias/análise , Espectrometria de Massas em Tandem , Compostos Orgânicos Voláteis/análise
14.
Chemosphere ; 251: 126390, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32146190

RESUMO

The aim of the study is to model concentrations of selected biogenic amines in various fish species (Atlantic salmon, Atlantic cod, striped catfish) bought in retail stores in Central Europe. Since the data contains non-detectable values, statistical methods for left-censored values from the exponential and Weibull distributions are applied and used to evaluate and compare the amount of biogenic amines in fish samples. Moreover, a risk of exceeding certain limits of biogenic amine concentrations to protect human health is determined. There are relatively high concentrations of putrescine, cadaverine and histamine in almost all fish species. Moreover, there was a significant difference in mean concentrations (distributions of concentrations, respectively) of histamine, tyramine and spermidine among the species.


Assuntos
Aminas Biogênicas/metabolismo , Peixes/metabolismo , Carne/análise , Animais , Aminas Biogênicas/química , Cadaverina/metabolismo , Monitoramento Ambiental , Europa (Continente) , Histamina/metabolismo , Humanos , Putrescina/metabolismo , Espermidina
15.
Sci Rep ; 10(1): 4116, 2020 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-32139747

RESUMO

Inflammatory mediators may have a role in various lower urinary tract disorders. Histamine is known to induce significant increases in both the tension and frequency of spontaneous phasic contractions in both urothelium with lamina propria (U&LP) and detrusor muscle via the activation of H1 receptor in juvenile animal models. However, it is unclear whether age affects these contractile responses to histamine. This study assessed the histamine receptor subtypes mediating contraction in juvenile and adult porcine bladders and compared the urothelium with lamina propria and detrusor responses to histamine. Isolated tissue bath studies were conducted using strips of porcine U&LP and detrusor obtained from juvenile (6 months) and adult (3 years) animals exposed to histamine receptor agonists and antagonists. Treatment with histamine (100 µM) in U&LP of juvenile animals caused increases in baseline tension by 47.84 ± 6.52 mN/g (p < 0.001, n = 51) and by 50.76 ± 4.10 mN/g (p < 0.001, n = 55) in adult animals. Furthermore, the frequency of spontaneous phasic contractions was significantly enhanced in response to histamine in U&LP of both juvenile and adult tissues (p < 0.001 for both age groups). Treatment with an H2 agonist in U&LP of juvenile animals decreased baseline tension by 13.97 ± 3.45 mN/g (n = 12, p < 0.05), but had no effect in adult animals. Inhibition of H1 receptors resulted in significantly reduced contractile responses of U&LP and detrusor to histamine in both juvenile and adult animals (p < 0.05). Treatment with an H2 receptor antagonist significantly enhanced contractions in juvenile preparations (n = 10, p < 0.05) but had no effect in adult preparations (n = 8). In detrusor, treatment with histamine (100 µM) in juvenile tissues showed a significantly higher increase in baseline tension of 19.10 ± 4.92 mN/g (n = 51) when compared to adult tissues exhibiting increases of 8.21 ± 0.89 mN/g (n = 56, p < 0.05). The increases in the baseline tension were significantly inhibited by the presence of H1 receptor antagonists in both juvenile and adult detrusor preparations. Treatment with either the H2 receptor antagonist or agonist in detrusor had no effect on both juvenile and adult tissues. Therefore, the histamine receptor system may play an essential role in the maintenance of bladder function or in bladder dysfunction observed in some lower urinary tract disorders.


Assuntos
Histamina/metabolismo , Membrana Mucosa/metabolismo , Receptores Histamínicos/metabolismo , Bexiga Urinária/metabolismo , Urotélio/metabolismo , Envelhecimento/metabolismo , Animais , Feminino , Histamina/farmacologia , Agonistas dos Receptores Histamínicos/farmacologia , Antagonistas dos Receptores Histamínicos , Técnicas In Vitro , Contração Muscular/efeitos dos fármacos , Suínos , Bexiga Urinária/efeitos dos fármacos , Urotélio/efeitos dos fármacos
16.
J Vis Exp ; (156)2020 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-32150162

RESUMO

Label-free impedance-based assays are increasingly used to non-invasively study ligand-induced GPCR activation in cell culture experiments. The approach provides real-time cell monitoring with a device-dependent time resolution down to several tens of milliseconds and it is highly automated. However, when sample numbers get high (e.g., dose-response studies for various different ligands), the cost for the disposable electrode arrays as well as the available time resolution for sequential well-by-well recordings may become limiting. Therefore, we here present a serial agonist addition protocol which has the potential to significantly increase the output of label-free GPCR assays. Using the serial agonist addition protocol, a GPCR agonist is added sequentially in increasing concentrations to a single cell layer while continuously monitoring the sample's impedance (agonist mode). With this serial approach, it is now possible to establish a full dose-response curve for a GPCR agonist from just one single cell layer. The serial agonist addition protocol is applicable to different GPCR coupling types, Gq Gi/0 or Gs and it is compatible with recombinant and endogenous expression levels of the receptor under study. Receptor blocking by GPCR antagonists is assessable as well (antagonist mode).


Assuntos
Bioensaio/métodos , Impedância Elétrica , Glioma/metabolismo , Histamina/metabolismo , Receptores Histamínicos/química , Receptores Histamínicos/metabolismo , Transdução de Sinais , Glioma/patologia , Humanos , Ligantes , Células Tumorais Cultivadas
17.
Nat Commun ; 11(1): 1397, 2020 03 13.
Artigo em Inglês | MEDLINE | ID: mdl-32170060

RESUMO

Gastrin-releasing peptide (GRP) functions as a neurotransmitter for non-histaminergic itch, but its site of action (sensory neurons vs spinal cord) remains controversial. To determine the role of GRP in sensory neurons, we generated a floxed Grp mouse line. We found that conditional knockout of Grp in sensory neurons results in attenuated non-histaminergic itch, without impairing histamine-induced itch. Using a Grp-Cre knock-in mouse line, we show that the upper epidermis of the skin is exclusively innervated by GRP fibers, whose activation via optogeneics and chemogenetics in the skin evokes itch- but not pain-related scratching or wiping behaviors. In contrast, intersectional genetic ablation of spinal Grp neurons does not affect itch nor pain transmission, demonstrating that spinal Grp neurons are dispensable for itch transmission. These data indicate that GRP is a neuropeptide in sensory neurons for non-histaminergic itch, and GRP sensory neurons are dedicated to itch transmission.


Assuntos
Peptídeo Liberador de Gastrina/genética , Peptídeo Liberador de Gastrina/metabolismo , Dor/metabolismo , Prurido/metabolismo , Células Receptoras Sensoriais/metabolismo , Medula Espinal/metabolismo , Animais , Modelos Animais de Doenças , Técnicas de Inativação de Genes , Histamina/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neurotransmissores , Dor/patologia , Prurido/patologia , Células Receptoras Sensoriais/patologia , Pele/metabolismo , Pele/patologia , Transcriptoma
18.
Int J Mol Sci ; 21(4)2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-32085403

RESUMO

The neurotransmitter levels of representatives from five different diagnosis groups were tested before and after participation in the MNRI®-Masgutova Neurosensorimotor Reflex Intervention. The purpose of this study was to ascertain neurological impact on (1) Developmental disorders, (2) Anxiety disorders/OCD (Obsessive Compulsive Disorder), PTSD (Post-Traumatic Stress disorder), (3) Palsy/Seizure disorders, (4) ADD/ADHD (Attention Deficit Disorder/Attention Deficit Disorder Hyperactive Disorder), and (5) ASD (Autism Spectrum Disorder) disorders. Each participant had a form of neurological dysregulation and typical symptoms respective to their diagnosis. These diagnoses have a severe negative impact on the quality of life, immunity, stress coping, cognitive skills, and social assimilation. This study showed a trend towards optimization and normalization of neurological and immunological functioning, thus supporting the claim that the MNRI method is an effective non-pharmacological neuromodulation treatment of neurological disorders. The effects of MNRI on inflammation have not yet been assessed. The resulting post-MNRI changes in participants' neurotransmitters show significant adjustments in the regulation of the neurotransmitter resulting in being calmer, a decrease of hypervigilance, an increase in stress resilience, behavioral and emotional regulation improvements, a more positive emotional state, and greater control of cognitive processes. In this paper, we demonstrate that the MNRI approach is an intervention that reduces inflammation. It is also likely to reduce oxidative stress and encourage homeostasis of excitatory neurotransmitters. MNRI may facilitate neurodevelopment, build stress resiliency, neuroplasticity, and optimal learning opportunity. There have been no reported side effects of MNRI treatments.


Assuntos
Inflamação/patologia , Neurotransmissores/metabolismo , Adolescente , Adulto , Transtorno do Deficit de Atenção com Hiperatividade/fisiopatologia , Transtorno do Deficit de Atenção com Hiperatividade/terapia , Transtorno do Espectro Autista/fisiopatologia , Transtorno do Espectro Autista/terapia , Dopamina/metabolismo , Epinefrina/biossíntese , Histamina/metabolismo , Humanos , Reflexo/fisiologia , Proteínas da Membrana Plasmática de Transporte de Serotonina/metabolismo , Adulto Jovem
19.
Lab Invest ; 100(6): 837-848, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32054995

RESUMO

Primary sclerosing cholangitis (PSC) is characterized by biliary damage and fibrosis. Multidrug resistance-2 gene knockout (Mdr2-/-) mice and PSC patients have increased histamine (HA) levels (synthesized by l-histidine decarboxylase, HDC) and HA receptor (HR) expression. Cholestatic HDC-/- mice display ameliorated biliary damage and hepatic fibrosis. The current study evaluated the effects of knockout of HDC-/- in Mdr2-/- mice (DKO) on biliary damage and hepatic fibrosis. WT, Mdr2-/- mice, and homozygous DKO mice were used. Selected DKO mice were treated with HA. We evaluated liver damage along with HDC expression and HA serum levels. Changes in ductular reaction were evaluated along with liver fibrosis, inflammation and bile acid signaling pathways. The expression of H1HR/PKC-α/TGF-ß1 and H2HR/pERK/VEGF-C was determined. In vitro, cholangiocyte lines were treated with HA with/without H1/H2 inhibitors before measuring: H1/H2HR, TGF-ß1, and VEGF-C expression. Knockout of HDC ameliorates hepatic damage, ductular reaction, fibrosis, inflammation, bile acid signaling and H1HR/PKC-α/TGF-ß1 and H2HR/pERK/VEGF-C signaling. Reactivation of the HDC/HA axis increased these parameters. In vitro, stimulation with HA increased HR expression and PKC-α, TGF-ß1, and VEGF-C expression, which was reduced with HR inhibitors. Our data demonstrate the key role for the HDC/HA axis in the management of PSC progression.


Assuntos
Colangite Esclerosante , Histamina/metabolismo , Histidina Descarboxilase , Cirrose Hepática , Transdução de Sinais/genética , Animais , Colangite Esclerosante/enzimologia , Colangite Esclerosante/metabolismo , Colangite Esclerosante/patologia , Modelos Animais de Doenças , Histidina Descarboxilase/genética , Histidina Descarboxilase/metabolismo , Fígado/citologia , Fígado/enzimologia , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/enzimologia , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Masculino , Camundongos , Camundongos Knockout
20.
PLoS One ; 15(2): e0229744, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32101568

RESUMO

Mast cells have secretory granules containing chemical mediators such as histamine and play important roles in the immune system. Polyamines are essential factors for cellular processes such as gene expression and translation. It has been reported that secretory granules contain both histamine and polyamines, which have similar chemical structures and are produced from the metabolism of cationic amino acids. We investigated the effect of polyamine depletion on mast cells using bone marrow-derived mast cells (BMMCs). Polyamine depletion was induced using α-difluoromethylornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase. DFMO treatment resulted in a significant reduction of cell number and abnormal secretory granules in BMMCs. Moreover, the cells showed a 2.3-fold increase in intracellular histamine and up-regulation of histidine decarboxylase (HDC) at the transcriptional level during BMMC differentiation. Levels of the transcription factor kruppel-like factor 4 (KLF4) greatly decreased upon DFMO treatment; however, Klf4 mRNA was expressed at levels similar to controls. We determined the translational regulation of KLF4 using reporter genes encoding Klf4-luc2 fusion mRNA, for transfecting NIH3T3 cells, and performed in vitro translation. We found that the efficiency of KLF4 synthesis in response to DFMO treatment was enhanced by the existence of a GC-rich 5'-untranslated region (5'-UTR) on Klf4 mRNA, regardless of the recognition of the initiation codon. Taken together, these results indicate that the enhancement of histamine synthesis by DFMO depends on the up-regulation of Hdc expression, achieved by removal of transcriptional suppression of KLF4, during differentiation.


Assuntos
Histamina/biossíntese , Fatores de Transcrição Kruppel-Like/metabolismo , Mastócitos/metabolismo , Animais , Medula Óssea/metabolismo , Células da Medula Óssea/metabolismo , Diferenciação Celular/fisiologia , Eflornitina/farmacologia , Feminino , Histamina/metabolismo , Histidina Descarboxilase/genética , Camundongos , Camundongos Endogâmicos C57BL , Células NIH 3T3 , Ornitina Descarboxilase/metabolismo , Poliaminas/metabolismo , Vesículas Secretórias/metabolismo , Espermidina/metabolismo
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