RESUMO
Histamine is a pleiotropic mediator involved in a broad spectrum of (patho)-physiological processes, one of which is the regulation of inflammation. Compounds acting on three out of the four known histamine receptors are approved for clinical use. These approved compounds comprise histamine H1-receptor (H1R) antagonists, which are used to control allergic inflammation, antagonists at H2R, which therapeutically decrease gastric acid release, and an antagonist at H3R, which is indicated to treat narcolepsy. Ligands at H4R are still being tested pre-clinically and in clinical trials of inflammatory diseases, including rheumatoid arthritis, asthma, dermatitis, and psoriasis. These trials, however, documented only moderate beneficial effects of H4R ligands so far. Nevertheless, pre-clinically, H4R still is subject of ongoing research, analyzing various inflammatory, allergic, and autoimmune diseases. During inflammatory reactions in gut tissues, histamine concentrations rise in affected areas, indicating its possible biological effect. Indeed, in histamine-deficient mice experimentally induced inflammation of the gut is reduced in comparison to that in histamine-competent mice. However, antagonists at H1R, H2R, and H3R do not provide an effect on inflammation, supporting the idea that H4R is responsible for the histamine effects. In the present review, we discuss the involvement of histamine and H4R in inflammatory and inflammation-associated diseases of the gut.
Assuntos
Gastroenteropatias/metabolismo , Gastroenteropatias/patologia , Inflamação/metabolismo , Inflamação/patologia , Receptores Histamínicos H4/metabolismo , Animais , Carcinogênese/metabolismo , Carcinogênese/patologia , Histamina/metabolismo , Humanos , Leucócitos/patologiaRESUMO
We hypothesized that, in mice, histamine via the histamine receptor subtype 4 (H4R) on colon epithelial cells affects epithelial barrier integrity, perturbing physiologic function of the colonic mucosa and thus aggravating the severity of colitis. To test this hypothesis, bone marrow-chimeric mice were generated from H4R knockout (H4R-/-) and wild-type (WT) BALB/cJ mice and subjected to the dextrane sodium sulfate (DSS)-induced acute colitis model. Clinical symptoms and pathohistological derangements were scored. Additionally, total RNA was extracted from either mouse whole-colon homogenates or primary cell preparations enriched for epithelial cells, and gene expression was analyzed by real-time quantitative polymerase chain reaction. The impact of the H4R on epithelial barrier function was assessed by measurement of transepithelial electrical resistence of organoid-derived two-dimensional monolayers from H4R-/- and WT mice using chopstick electrodes. Bone marrow-chimeric mice with genetic depletion of the H4R in nonhematopoietic cells exhibited less severe DSS-induced acute colitis symptoms compared with WT mice, indicating a functional proinflammatory expression of H4R in nonimmune cells of the colon. Analysis of H4R expression revealed the presence of H4R mRNA in colon epithelial cells. This expression could be confirmed and complemented by functional analyses in organoid-derived epithelial cell monolayers. Thus, we conclude that the H4R is functionally expressed in mouse colon epithelial cells, potentially modulating mucosal barrier integrity and intestinal inflammatory reactions, as was demonstrated in the DSS-induced colitis model, in which presence of the H4R on nonhematopoietic cells aggravated the inflammatory phenotype. SIGNIFICANCE STATEMENT: The histamine H4 receptor (H4R) is functionally expressed on mouse colon epithelial cells, thereby aggravating dextrane sodium sulfate-induced colitis in BALB/cJ mice. Histamine via the H4R reduces transepithelial electrical resistance of colon epithelial monolayers, indicating a function of H4R in regulation of epithelial barrier integrity.
Assuntos
Colo/fisiologia , Mucosa Intestinal/fisiologia , Receptores Histamínicos H4/fisiologia , Animais , Colite/induzido quimicamente , Sulfato de Dextrana , Impedância Elétrica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , RNA Mensageiro/análise , Receptores Histamínicos H4/genéticaRESUMO
Histamine is a pro-inflammatory mediator with a prominent role in allergic diseases. Antagonists at the histamine receptor subtype 1 are central in anti-allergic therapies, with the exception of allergic asthma, where they are clinically without effect. The latest identified histamine receptor subtype 4, which is expressed mainly in hematopoietic cells, now provides a reasonable target for new therapeutic strategies inhibiting histamine function. The pathophysiology of allergy, esp. allergic asthma, and in its context the effects of antagonists at the histamine receptor subtype 4 in preclinical and clinical settings are discussed in this chapter.
Assuntos
Asma/metabolismo , Receptores Histamínicos/metabolismo , Animais , Asma/tratamento farmacológico , Antagonistas dos Receptores Histamínicos/farmacologia , Antagonistas dos Receptores Histamínicos/uso terapêutico , Humanos , Hipersensibilidade/tratamento farmacológico , Hipersensibilidade/metabolismoRESUMO
Via the histamine H4 -receptor (H4 R), histamine promotes the pathogenesis of experimental allergic asthma in mice. Application of H4 R antagonists during sensitization as well as during provocation reduces the severity of the disease. However, the specific cell types functionally expressing H4 R in experimental allergic asthma have not been well characterized in vivo. In this study, we identified the cell type(s) responsible for H4 R activity in experimental asthma and related physiological mechanisms. Using H4 R-deficient mice, we studied the role of H4 R in the sensitization and effector phase. DCs lacking H4 R expression during the in vitro sensitization reaction resulted in effector T cells unable to induce an entire eosinophilic inflammation in the lung upon adoptive transfer in vivo. Recipient mice lacking H4 R expression, which were adoptively transferred with H4 R(+/+) T cells polarized in the presence of H4 R(+/+) DCs, showed reduced signs of inflammation and ameliorated lung function. Here, we provide in vivo evidence that in experimental asthma in mice the H4 R specifically regulates activation of DCs during sensitization, while in the effector phase the H4 R is active in cells involved in the activation of eosinophils, and possibly other cells. A putative therapy targeting the H4 R may be an option for asthma patients developing IL-5-dependent eosinophilia.
Assuntos
Asma/imunologia , Células Dendríticas/imunologia , Eosinófilos/imunologia , Inflamação/imunologia , Receptores Acoplados a Proteínas G/imunologia , Receptores Histamínicos/imunologia , Transferência Adotiva , Alérgenos/imunologia , Animais , Asma/induzido quimicamente , Asma/patologia , Líquido da Lavagem Broncoalveolar/citologia , Antígeno CD11c/metabolismo , Citocinas/imunologia , Modelos Animais de Doenças , Histamina/metabolismo , Interleucina-5/imunologia , Pulmão/imunologia , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Ovalbumina , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Acoplados a Proteínas G/genética , Receptores Histamínicos/genética , Receptores Histamínicos H4 , Células Th2/imunologia , Células Th2/transplanteRESUMO
Clostridium difficile may induce antibiotic-associated diarrhoea and, in severe cases, pseudomembranous colitis characterized by tremendous neutrophil infiltration. All symptoms are caused by two exotoxins: TcdA and TcdB. We describe here the activation of isolated human blood neutrophils by TcdB and, moreover, by toxin fragments generated by limited proteolytical digestion. Kinetics and profiles of TcdB-induced rise in intracellular-free Ca(2+) and reactive oxygen species production were similar to that induced by fMLF, which activates the formyl peptide receptor (FPR) recognizing formylated bacterial peptide sequences. Transfection assays with the FPR-1 isoform hFPR26 in HEK293 cells, heterologous desensitization experiments and FPR inhibition via cyclosporine H strongly suggest activation of cells via FPR-1. Domain analyses revealed that the N-terminal glucosyltransferase domain of TcdB is a potent activator of FPR pointing towards an additional mechanism that might contribute to pathogenesis. This pro-inflammatory ligand effect can be triggered even by cleaved and, thus, non-cytotoxic toxin. In summary, we report (i) a ligand effect on neutrophils as completely new molecular mode of action, (ii) pathogenic potential of truncated or proteolytically cleaved 'non-cytotoxic' fragments and (iii) an interaction of the N-terminal glucosyltransferase domain instead of the C-terminal receptor binding domain of TcdB with target cells.
Assuntos
Proteínas de Bactérias/metabolismo , Toxinas Bacterianas/metabolismo , Clostridioides difficile/imunologia , Neutrófilos/imunologia , Neutrófilos/microbiologia , Fragmentos de Peptídeos/metabolismo , Receptores de Formil Peptídeo/metabolismo , Células HEK293 , HumanosRESUMO
OBJECTIVE AND DESIGN: The modulation of antigen uptake and activation of dendritic cells (DCs) by histamine may function as a regulator of inflammation. Therefore, we sought to determine the impact of histamine on antigen uptake by and activation of murine DCs. MATERIAL AND METHODS: DCs from spleen and lung were either identified by flow cytometry or were immunomagnetically enriched. Cells were stimulated with histamine, and the regulation of MHC-II and co-stimulatory molecule expression (CD80, CD86, and ICOS-L) and antigen uptake were quantified by flow cytometry. Individual contributions of the histamine receptor subtypes were determined by using the antagonists mepyramine (histamine H1-receptor: H1R), famotidine (H2R), and JNJ 7777120 (H4R). RESULTS: Histamine accelerated the uptake of soluble antigen via the H1R, H2R, and H4R in splenic DCs. Co-stimulatory molecule expression was enhanced already by enrichment procedures, thus, the analyses were performed in unseparated cell populations. Histamine enhanced the expression of CD86 and ICOS-L while expression of CD80 was unaffected. Antagonism at H1R, H2R, and H4R and at H1R and H4R reduced the histamine-induced enhanced expression of CD86 and ICOS-L, respectively. CONCLUSIONS: Histamine contributes to the regulation of the immunological synapse by stimulation of antigen uptake and activation of DCs via H1R, H2R, and H4R.
Assuntos
Células Dendríticas/imunologia , Histamina/farmacologia , Sinapses Imunológicas/imunologia , Animais , Antígeno B7-1/imunologia , Antígeno B7-2/imunologia , Células Cultivadas , Células Dendríticas/citologia , Ligante Coestimulador de Linfócitos T Induzíveis/imunologia , Camundongos , Receptores Histamínicos H1/imunologiaRESUMO
The histamine H4 receptor (H4R) is a classic pertussis toxin-sensitive Gi protein-coupled receptor that mediates increases in intracellular calcium concentration ([Ca(2+)]i). The presence of H4R in human eosinophils has been rigorously documented by several independent groups. It has also been suggested that H4R is expressed in human monocytes, but this suggestion hinges in part on H4R antibodies with questionable specificity. This situation prompted us to reinvestigate H4R expression in human monocytes. As positive control, we studied human embryonic kidney 293T cells stably expressing the human H4R (hH4R). In these cells, histamine (HA) and the H4R agonist UR-PI376 (2-cyano-1-[4-(1H-imidazol-4-yl)butyl]-3-[(2-phenylthio)ethyl]guanidine) induced pertussis toxin-sensitive [Ca(2+)]i increases. However, in quantitative real-time polymerase chain reaction studies we failed to detect hH4R mRNA in human monocytes and U937 promonocytes. In human monocytes, ATP and N-formyl-l-methionyl-l-leucyl-l-phenylalanine increased [Ca(2+)]i, but HA, UR-PI376, and 5-methylhistamine (a dual H4R/H2 receptor agonist) did not. In U937 promonocytes and differentiated U937 cells, HA increased [Ca(2+)]i, but this increase was mediated via HA H1 receptor. In conclusion, there is no evidence for the presence of H4R in human monocytes.
Assuntos
Monócitos/metabolismo , Receptores Acoplados a Proteínas G/biossíntese , Receptores Histamínicos/biossíntese , Adulto , Guanidinas/farmacologia , Células HEK293 , Agonistas dos Receptores Histamínicos/farmacologia , Antagonistas dos Receptores Histamínicos/farmacologia , Humanos , Imidazóis/farmacologia , Masculino , Monócitos/efeitos dos fármacos , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Histamínicos H4 , Células U937RESUMO
The diverse functions of histamine are mediated by four specific histamine receptor subtypes, which belong to the family of G-protein-coupled receptors. Here, we summarize data obtained with histamine-deficient L-histidine decarboxylase knockout and histamine receptor subtype knockout mice in inflammation models. Advantages and disadvantages of the knockout approaches compared with pharmacologic approaches are discussed critically. Due to many controversial data it is very difficult to draw clear-cut conclusions from the data provided in the literature. Thus, the published studies highlight the complexity of histamine function in inflammation and the need for much more systematic experimental work.
Assuntos
Modelos Animais de Doenças , Inflamação/genética , Inflamação/metabolismo , Receptores Histamínicos/deficiência , Receptores Histamínicos/metabolismo , Animais , Humanos , Inflamação/patologia , Camundongos , Camundongos Knockout , Receptores Histamínicos/genéticaRESUMO
OBJECTIVE: Expression and function of histamine H4-receptor, an immunomodulatory receptor involved in inflammatory diseases, on murine macrophages, which are vital for immunity, were investigated. MATERIALS AND METHODS: The expression pattern of histamine receptors on bone marrow-derived macrophages of BALB/c mice and on RAW 264.7 cells was studied at the mRNA level by reverse transcription polymerase chain reaction. The functional relevance of histamine receptors was investigated by analyzing histamine-induced chemotaxis and phagocytosis in the presence of histamine receptor antagonists mepyramine (histamine H1-receptor), famotidine (histamine H2-receptor), thioperamide (histamine H3/4-receptors) and JNJ7777120 (histamine H4-receptor). RESULTS: Both bone marrow-derived macrophages and RAW 264.7 cells express mRNA for histamine H1-receptor and histamine H4-receptor. Residual amounts of histamine H2-receptor mRNA are found in bone marrow-derived macrophages only. In both cellular models, histamine induced chemotaxis and phagocytic activity, which was reduced by thioperamide as well as by JNJ 7777120, but not by mepyramine or famotidine. CONCLUSION: In murine bone marrow-derived macrophages and RAW 264.7 macrophage-like cells histamine H4-receptor mediates chemotaxis and phagocytic activity.
Assuntos
Células da Medula Óssea/efeitos dos fármacos , Quimiotaxia/efeitos dos fármacos , Histamina/farmacologia , Macrófagos/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Receptores Acoplados a Proteínas G/efeitos dos fármacos , Receptores Histamínicos/efeitos dos fármacos , Animais , Células da Medula Óssea/metabolismo , Linhagem Celular , Relação Dose-Resposta a Droga , Feminino , Citometria de Fluxo , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase em Tempo Real , Receptores Acoplados a Proteínas G/biossíntese , Receptores Histamínicos/biossíntese , Receptores Histamínicos H4RESUMO
In previous studies, we demonstrated the involvement of H4R in inflammatory bowel disease (IBD) and IBD-associated colon cancer in mice and could ascribe H4R-mediated histamine function to colon epithelial cells. The transferability of obtained data to humans is however lacking. Functional expression of H4R on colon epithelial cells is a prerequisite to pursue the hypothesis of involvement of H4R in carcinogenesis. Thus, we here compared the expression of histamine receptor subtypes in a series of cell lines. Out of these, three colon-derived cell lines displaying different combinations of H1R and H4R expression were submitted to functional analyses. Human hematopoietic HMC-1, HL-60, and U937, lung-derived A549 and Calu-3, and colorectal LoVo, SW 480, Caco-2, HT-29, and HCT116 cells were included in the study. mRNA expression was quantified by RT-qPCR. For functional analyses, Caco-2, HT-29, and HCT116 cells were treated by incubation with 1 - 10 µM histamine in the presence or absence of selective histamine receptor antagonists. Calcium mobilization, cAMP accumulation, and cell proliferation were measured by fluorimetry, mass spectrometry, and real-time bioimpedance measurements, respectively. Histamine receptor expression was heterogeneous in the cell lines tested. In most cell lines, we detected H1R mRNA while H4R mRNAs were found only occasionally. The colon-derived epithelial cell lines LoVo, SW480, and HT-29 expressed H1R mRNA exclusively, while in HCT116 cells H1R and H4R mRNAs and in CaCo-2 H2R mRNA were detectable. Subsequent functional analyses in HT29, Caco-2, and HCT116 cells, however, indicated that only HT-29 responded to histamine stimulation, by means of H1R. For a detailed analysis of histamine receptor function, esp. that of H1R and H4R, in human colon-derived cell lines, the cell lines tested here are not fully convenient unless genetically modified.
Assuntos
Histamina , Doenças Inflamatórias Intestinais , Humanos , Camundongos , Animais , Histamina/farmacologia , Histamina/metabolismo , Receptores Histamínicos H1/genética , Receptores Histamínicos H1/metabolismo , Receptores Histamínicos H4 , Células CACO-2 , Receptores Histamínicos/genética , Receptores Histamínicos/metabolismo , Colo/metabolismo , RNA MensageiroRESUMO
The histamine H(4) receptor (H(4)R) is expressed in several cell types of the immune system and is assumed to play an important pro-inflammatory role in various diseases, including bronchial asthma, atopic dermatitis, and pruritus. Accordingly, H(4)R antagonists have been suggested to provide valuable drugs for the treatment of these diseases. Over the past decade, the indole derivative 1-[(5-chloro-1H-indol-2-yl)carbonyl]-4-methylpiperazine (JNJ7777120) has become the "standard" H(4)R antagonist and has been extensively used to assess the pathophysiological role of the H(4)R. However, the situation has now become more complicated by recent data (p. 749 and Naunyn Schmiedebergs Arch Pharmacol doi: 10.1007/s00210-011-0612-3) showing that JNJ7777120 can also activate ß-arrestin in a supposedly G(i)-protein-independent (pertussis toxin-insensitive) manner and that at certain H(4)R species orthologs, JNJ7777120 exhibits partial agonist efficacy with respect to G(i)-protein activation (steady-state high-affinity GTPase activity). These novel findings can be explained within the concept of functional selectivity or biased signaling, assuming unique ligand-specific receptor conformations with distinct signal transduction capabilities. Thus, great caution must be exerted when interpreting in vivo effects of JNJ7777120 as H(4)R antagonism. We discuss future directions to get out of the current dilemma in which there is no "standard" H(4)R antagonist available to the scientific community.
Assuntos
Antagonistas dos Receptores Histamínicos/farmacologia , Indóis/farmacologia , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/fisiologia , Piperazinas/farmacologia , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Acoplados a Proteínas G/química , Receptores Histamínicos/química , Animais , Arrestinas/metabolismo , Humanos , Proteínas de Membrana/metabolismo , Estrutura Terciária de Proteína/efeitos dos fármacos , Estrutura Terciária de Proteína/fisiologia , Receptores Acoplados a Proteínas G/metabolismo , Receptores Histamínicos/metabolismo , Receptores Histamínicos H4 , beta-ArrestinasRESUMO
OBJECTIVE: Histamine plays a role in several (patho) physiological processes that are commonly studied in mouse models. However, a systematic quantification of histamine and its metabolite N-methylhistamine in mouse organs has not been reported so far. METHODS: Balb/c and C57Bl/6 mice were grouped according to their sex and age. Brains, hearts, lungs, livers, kidneys, stomachs, intestines, thymi, spleens, and lymph nodes were excised, weighed, and homogenized. Histamine and N-methylhistamine were quantified simultaneously by a HPLC-mass spectrometry method. RESULTS: In all organs analyzed, histamine and N-methylhistamine were detected; however, with quantitative differences. Histamine was present most abundantly in the stomach, lymph nodes, and thymus. The lowest histamine concentrations were detected in brain, liver, lung, and intestine. In most organs, the histamine concentrations increased age-dependently. Substantial concentrations of N-methylhistamine were detected only in lung, intestine and kidney, while in all other organs it was present only in minor quantities. CONCLUSION: HPLC-mass spectrometry is a useful method for the highly sensitive and simultaneous detection of histamine and N-methylhistamine. Histamine is present in virtually all organs, not only in those traditionally associated with histamine-mediated disease. Highest concentrations are found in stomach, lymph node, and thymus; medium concentrations in heart, spleen, and kidney; and lowest concentrations detected in intestine, brain, liver, and lung.
Assuntos
Histamina/análise , Metilistaminas/análise , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Animais , Química Encefálica , Cromatografia Líquida de Alta Pressão , Feminino , Intestinos/química , Rim/química , Fígado/química , Pulmão/química , Linfonodos/química , Masculino , Camundongos , Miocárdio/química , Baço/química , Estômago/química , Espectrometria de Massas em Tandem , Timo/químicaRESUMO
Administration of the multipotent hematopoietic progenitor cell (HPC) line DKmix improved cardiac function after myocardial infarction and accelerated dermal wound healing due to paracrine mechanisms. The aim of this study was to analyse the secreted proteins of DKmix cells in order to identify the responsible paracrine factors and assess their relevance to the wide spectrum of therapeutic effects. A mass spectrometry (MS)-based approach was used to identify secreted proteins of DKmix cells. Serum free culture supernatants of DKmix-conditioned medium were collected and the proteins present were separated, digested by trypsin and the resulting peptides were then analyzed by matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) MS. Overall 95 different proteins were identified. Among them, secretory proteins galectin-3 and gelsolin were identified. These proteins are known to stimulate cell migration and influence wound healing and cardiac remodelling. The remaining proteins originate from intracellular compartments like cytoplasm (69%), nucleus (12%), mitochondria (4%), and cytoplasmic membrane (3%) indicating permeable or leaky DKmix cells in the conditioned medium. Additionally, a sandwich immunoassay was used to detect and quantify cytokines and chemokines. Interleukin-6 (IL-6), interleukin-13 (IL-13), monocyte-chemoattractant protein-1 (MCP-1), monocyte-chemoattractant protein-3 (MCP-3), monocyte-chemoattractant protein-1alpha (MIP-1alpha) and monocyte-chemoattractant protein-1beta (MIP-1beta) were detected in low concentrations. This study identified a subset of proteins present in the DKmix-conditioned medium that act as paracrine modulators of tissue repair. Moreover, it suggests that DKmix-derived conditioned medium might have therapeutic potency by promoting tissue regeneration.
Assuntos
Espectrometria de Massas/métodos , Células-Tronco Multipotentes/metabolismo , Proteoma/química , Animais , Linhagem Celular/metabolismo , Movimento Celular/efeitos dos fármacos , Meios de Cultivo Condicionados , Citocinas/química , Fibroblastos/metabolismo , Camundongos , Células-Tronco Multipotentes/citologia , Comunicação Parácrina , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/classificação , Proteoma/metabolismo , Proteoma/farmacologia , Frações Subcelulares/químicaRESUMO
The histamine H4 receptor is expressed preferentially on immune cells, indicating a possible role of the H4 receptor in inflammation. Studies of inflammation in several animal models point to a pro-inflammatory function of the H4 receptor. However, studies on experimental murine bronchial asthma yielded conflicting results, a fact which is neglected in most H4 receptor publications. Therefore, the present review critically analyzes available data on the role of the H4 receptor in the murine bronchial asthma model.
Assuntos
Asma/fisiopatologia , Receptores Acoplados a Proteínas G/fisiologia , Receptores Histamínicos/fisiologia , Animais , Modelos Animais de Doenças , Histamina/fisiologia , Humanos , Indóis/farmacologia , Inflamação/fisiopatologia , Camundongos , Piperazinas/farmacologia , Receptores Acoplados a Proteínas G/efeitos dos fármacos , Receptores Histamínicos/efeitos dos fármacos , Receptores Histamínicos H3/fisiologia , Receptores Histamínicos H4RESUMO
BACKGROUND: Anorexia, body wasting, inflammation, muscle, and adipose tissue loss are hallmarks of cancer cachexia, a syndrome that affects the majority of cancer patients, impairing their ability to endure chemotherapeutic therapies and reducing their lifespan. In the last 10 years, alterations of protein turnover and impairment of adult myogenesis have been proposed as major contributing factors. METHODS: Muscle stem cells, including satellite cells, mesoangioblasts, and fibroadipogenic progenitors, were isolated and characterized from C26 colon carcinoma-bearing (C26) mice. Circulating levels of interleukin-4/13 (IL4/IL13) were analysed by ELISA, and the effects of IL4 on muscle mass and function, protein synthesis, muscle regeneration, and myogenic progenitor cell number were analysed at both functional (treadmill and grip test) and molecular levels (qRT-PCR, immunofluorescence analysis, surface sensing of translation, and western blot). The Kaplan-Meier test was used to analyse the survival curve of IL4-treated and IL4-untreated C26 mice. RESULTS: The administration of IL4 to C26 mice rescued muscle mass by increasing protein synthesis. The IL4 treatment improved performances and prolonged survival of C26 mice. IL4 administration re-established both number and function of satellite cells and fibroadipogenic progenitors without affecting mesoangioblasts in C26 mice, rescuing myogenesis. Upon IL4 treatment, a high number of cytotoxic lymphocytes and type II macrophages were observed with a subsequent increase in necrotic areas of C26 tumours. CONCLUSIONS: The results here presented shed new light on IL4 signalling during muscle wasting and early stages of muscle regeneration that explain the beneficial effect observed in IL4-treated C26 mice. These findings might aid to develop therapeutic approaches to improve mobility and quality of life in cachectic patients.
Assuntos
Neoplasias do Colo/genética , Neoplasias do Colo/metabolismo , Interleucina-4/uso terapêutico , Desenvolvimento Muscular/genética , Músculo Esquelético/metabolismo , Animais , Humanos , Interleucina-4/farmacologia , Camundongos , RatosRESUMO
Colorectal cancer (CRC), a severe complication of inflammatory bowel diseases, is a common type of cancer and accounts for high mortality. CRC can be modeled in mice by application of the tumor promoter, azoxymethane (AOM), in combination with dextran sodium sulfate (DSS), which are able to induce colitis-like manifestations. Active colitis correlates with high mucosal concentrations of histamine, which, together with the histamine receptor subtype 4 (H4R), provide a pro-inflammatory function in a mouse colitis model. Here, we analyzed whether H4R is involved in the pathogenesis of AOM/DSS-induced CRC in mice. As compared to wild type (WT) mice, AOM/DSS-treated mice lacking H4R expression (TM) demonstrate ameliorated signs of CRC, i.e., significantly reduced loss of body weight, stiffer stool consistency, and less severe perianal bleeding. Importantly, numbers and diameters of tumors and the degree of colonic inflammation are dramatically reduced in TM mice as compared to WT mice. This is concomitant with a reduced colonic inflammatory response involving expression of cyclooxygenase 2 and the production of C-X-C motif chemokine ligand 1 (CXCL1) and CXCL2. We conclude that H4R is involved in the tumorigenesis of chemically-induced CRC in mice via cyclooxygenase 2 expression and, probably, CXCL1 and CXCL2 as effector molecules.
RESUMO
The human histamine H(4)-receptor (hH(4)R) possesses high constitutive activity and, like the human H(1)-receptor (hH(1)R), is involved in the pathogenesis of type-I allergic reactions. The study aims were to explore the value of dual H(1)/H(4)R antagonists as antiallergy drugs and to address the question of whether H(1)R ligands bind to hH(4)R. In an acute murine asthma model, the H(1)R antagonist mepyramine and the H(4)R antagonist 1-[(5-chloro-1H-indol-2-yl)carbonyl]-4-methyl-piperazine (JNJ 7777120) exhibited synergistic inhibitory effects on eosinophil accumulation in the bronchoalveolar lavage fluid. At the hH(4)R expressed in Sf9 insect cells, 18 H(1)R antagonists and 22 H(1)R agonists showed lower affinity to hH(4)R than to hH(1)R as assessed in competition binding experiments. For a small number of H(1)R antagonists, hH(4)R partial agonism was observed in the steady-state GTPase assay. Most compounds were neutral antagonists or inverse agonists. Twelve phenylhistamine-type hH(1)R partial agonists were also hH(4)R partial agonists. Four histaprodifen-type hH(1)R partial agonists were hH(4)R inverse agonists. Dimeric histaprodifen was a more efficacious hH(4)R inverse agonist than the reference compound thioperamide. Suprahistaprodifen was the only histaprodifen acting as hH(4)R partial agonist. Suprahistaprodifen was docked into the binding pocket of inactive and active hH(4)R models in two different orientations, predominantly stabilizing the active state of hH(4)R. Collectively, the synergistic effects of H(1)R and H(4)R antagonists in an acute asthma model and the overlapping interaction of structurally diverse H(1)R ligands with hH(1)R and hH(4)R indicate that the development of dual H(1)R/H(4)R antagonists is a worthwhile and technically feasible goal for the treatment of type-I allergic reactions.
Assuntos
Agonistas dos Receptores Histamínicos/metabolismo , Antagonistas dos Receptores Histamínicos H1/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Receptores Histamínicos/metabolismo , Sequência de Aminoácidos , Animais , Células Cultivadas , Feminino , Histamina/análogos & derivados , Histamina/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Modelos Moleculares , Dados de Sequência Molecular , Ligação Proteica/fisiologia , Receptores Acoplados a Proteínas G/genética , Receptores Histamínicos/genética , Receptores Histamínicos H4 , Homologia de Sequência de Aminoácidos , SpodopteraRESUMO
Ligand binding in the TLR/IL-1R family results in the transient formation of an intracellular signaling complex, which contains, amongst others, the serine/threonine-specific kinase IL-1R-associated kinase 1 (IRAK-1). Concomitantly, the kinase function of IRAK-1 becomes activated, resulting in massive autophosphorylation and finally in the dissociation of the initially constituted signaling complex. The death domain (DD) of IRAK-1 mediates the interaction with other molecules of the signaling complex, e.g., the adaptor MyD88, the silencer Tollip, and the activator kinase IRAK-4. The conserved threonine at position 66 (T66), located within the DD, is a putative autophosphorylation target site. Here, we provide evidence that T66 critically impacts the secondary structure of the IRAK-1 DD. Thereby, it ensures the transient manner of interactions between IRAK-1 and the other signaling molecules. This essential role, however, is not regulated by phosphorylation of T66 itself.
Assuntos
Quinases Associadas a Receptores de Interleucina-1/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Fator 88 de Diferenciação Mieloide/metabolismo , Treonina/química , Células Cultivadas , Transferência Ressonante de Energia de Fluorescência , Vetores Genéticos , Humanos , Immunoblotting , Imunoprecipitação , Quinases Associadas a Receptores de Interleucina-1/química , Quinases Associadas a Receptores de Interleucina-1/genética , Interleucina-2/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/genética , Rim/metabolismo , Peso Molecular , Fator 88 de Diferenciação Mieloide/genética , NF-kappa B/genética , NF-kappa B/metabolismo , Fosforilação , Transdução de Sinais , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Treonina/genética , Treonina/metabolismoRESUMO
T cells and T cell derived cytokines are involved in the complex pathogenesis of asthma. The role of the cytokine IL-18 however, is not clearly defined so far. On the one hand side IL-18 induces Th1-type cytokines and thereby might counter-regulate Th2-mediated allergic asthma. On the other hand IL-18 also bears pro-inflammatory effects possibly enhancing experimental asthma. In order to elucidate the role of IL-18 in allergic pulmonary inflammation typical symptoms were compared after induction of experimental asthma in IL-18(-/-) and in wild type mice. Asthma was induced using ovalbumin (OVA) as allergen for sensitization and challenge. Sham sensitized and OVA challenged mice served as controls. Bronchoalveolar lavage-fluid cytology, leukocyte infiltration in lung tissues, serum levels of OVA-specific IgE and cytokines, and lung function were analyzed. Clear differences could be observed between control and asthmatic mice, both in wild type and IL-18(-/-) animals. Surprisingly, no differences were found between asthmatic wild type and IL-18(-/-) mice. Thus, in contrast to conflicting data in the literature IL-18 did not suppress or enhance the pulmonary allergic immune response in a murine experimental model of asthma.
Assuntos
Asma/imunologia , Interleucina-18/sangue , Animais , Asma/patologia , Testes de Provocação Brônquica , Líquido da Lavagem Broncoalveolar , Modelos Animais de Doenças , Eosinófilos/imunologia , Feminino , Imunização , Imunoglobulina E/sangue , Interleucina-17/sangue , Interleucina-5/sangue , Contagem de Leucócitos , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Ovalbumina/farmacologia , Pletismografia TotalRESUMO
Sialylation of glycans is ubiquitous in vertebrates, but was believed to be absent in plants, arthropods, and fungi. However, recently evidence has been provided for the presence of sialic acid in these evolutionary clades. In addition, homologs of mammalian genes involved in sialylation can be found in the genomes of these taxa and for some Drosophila enzymes, involvement in sialic acid metabolism has been documented. In plant genomes, homologs of sialyltransferase genes have been identified, but there activity could not be confirmed. Several mammalian cell lines exist with defects in the sialylation pathway. One of these is the Chinese hamster ovary cell line Lec2, deficient in CMP-sialic acid transport to the Golgi lumen. These mutants provide the possibility to clone genes by functional complementation. Using expression cloning, we have identified an Arabidopsis thaliana nucleotide sugar transporter that is able to complement the CMP-sialic acid transport deficiency of Lec2 cells. The isolated gene (At5g41760) is a member of the triose-phosphate/nucleotide sugar transporter gene family. Recombinant expression of the gene in yeast and testing in vitro confirmed its ability to transport CMP-sialic acid.