RESUMO
BACKGROUND: Little is known whether sublingual immunotherapy using Japanese cedar pollen extract (cedar SLIT) is effective for not only Japanese cedar pollinosis but also Japanese cypress pollinosis. We investigated the prevalence rate of Japanese cypress pollinosis, efficacy of cedar SLIT on cypress pollinosis and patients' wish to receive cypress SLIT. METHODS: We investigated a multi-center (31 institutions), cross-sectional survey using a self-administrated questionnaire with four questions for patients received cedar SLIT aged from 5 to 69 years old. RESULTS: 2523 subjects were enrolled for analysis. 83.4% of them had pollinosis symptoms during cypress season before cedar SLIT. In such patients, 37.4% experienced lessened efficacy of cedar SLIT during cypress season. Both the prevalence of cypress pollinosis and the lessened efficacy of cedar SLIT on cypress pollinosis were significantly seen in western Japan as compared to eastern Japan. 76.1% of the subject having cypress pollinosis before SLIT wished to receive cypress SLIT if it is available. CONCLUSION: A lessened efficacy of cedar SLIT during cypress season was broadly seen in Japan, and further showed a regional difference. Together with the finding of high wish by patients, these results suggest a development of cypress SLIT is desirable.
Assuntos
Cryptomeria , Cupressus , Rinite Alérgica Sazonal , Imunoterapia Sublingual , Humanos , Pré-Escolar , Criança , Adolescente , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Idoso , Rinite Alérgica Sazonal/terapia , Rinite Alérgica Sazonal/tratamento farmacológico , Pólen , Estudos Transversais , Prevalência , Inquéritos e Questionários , AlérgenosRESUMO
BACKGROUND: Clara cell 10-kD protein (CC10) is well known to be an immuno-suppressive protein secreted from airway epithelial cells after inflammatory stimulation and is involved in the development of allergic disorders. Although histamine H1 receptor antagonists are used for the treatment of allergic disorders, the influence of the agents on CC10 production is not well understood. In the present study, we examined the influence of a histamine H1 receptor antagonist, fexofenadine hydrochloride (FEX) on CC10 production in vitro and in vivo. METHODS: Nasal epithelial cells (5 x 10(6) cells/ml) were stimulated with 20 ng/ml TNF-alpha in the presence of various concentrations of FEX for 24 hours. CC10 levels in culture supernatants were examined by ELISA. Patients with Japanese cedar pollinosis were treated orally with FEX twice a day at a single dose of 60 mg for two weeks during Japanese cedar pollen season (February 2011 to April 2011). CC10 levels in nasal secretions were also examined by ELISA. RESULTS: The addition of FEX into cell cultures caused increase in CC10 production induced by TNF-alpha stimulation, and the minimum concentration that caused significant increase was 200 ng/ml. Oral administration of FEX also increased CC10 levels in nasal secretions from pollinosis patients along with attenuation of clinical symptoms. CONCLUSION: The ability of FEX to enhance CC10 production may account, at least in part, for the clinical efficacy of the agent in allergic disorders, including allergic rhinitis.
Assuntos
Células Epiteliais/imunologia , Antagonistas dos Receptores Histamínicos H1/uso terapêutico , Cavidade Nasal/imunologia , Rinite Alérgica Sazonal/tratamento farmacológico , Terfenadina/análogos & derivados , Uteroglobina/biossíntese , Adulto , Células Cultivadas , Cryptomeria/imunologia , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Antagonistas dos Receptores Histamínicos H1/farmacologia , Humanos , Masculino , Pessoa de Meia-Idade , Cavidade Nasal/citologia , Cavidade Nasal/efeitos dos fármacos , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Rinite Alérgica Sazonal/patologia , Índice de Gravidade de Doença , Terfenadina/farmacologia , Terfenadina/uso terapêutico , Fator de Necrose Tumoral alfa/farmacologia , Uteroglobina/imunologia , Uteroglobina/metabolismoRESUMO
Dysregulated expression of Maf proteins (namely c-Maf, MafA and MafB) leads to multiple myeloma in humans and oncogenic transformation of chicken embryonic fibroblasts. Maf proteins are transcriptional activators of tissue-specific gene expression and regulators of cell differentiation. For example, MafA is a critical regulator of crystallin genes and the lens differentiation program in chickens. In mammals, MafA is essential for the development of mature insulin-producing beta-cells of pancreas. It has been shown that MafA protein stability is regulated by phosphorylations at multiple serine and threonine residues. Here, we report that Maf proteins are also post-translationally modified by small ubiquitin-like modifier (SUMO) proteins at a conserved lysine residue in the amino-terminal transactivator domain. A SUMOylation-deficient mutant of MafA (K32R) was more potent than wild-type MafA in transactivating luciferase reporter construct driven by alphaA-crystallin or insulin gene promoter. In ovo electroporation into developing chicken embryo showed that the K32R mutant induced ectopic delta-crystallin gene expression more efficiently than the wild-type MafA. We also demonstrated that the K32R mutant had enhanced ability to induce colony formation of a chicken fibroblast cell line DF-1. Therefore, SUMOylation is a functional post-translational modification of MafA that negatively regulates its transcriptional and transforming activities.
Assuntos
Transformação Celular Neoplásica/genética , Fatores de Transcrição Maf Maior/genética , Sumoilação , Transcrição Gênica/genética , Animais , Linhagem Celular , Linhagem Celular Tumoral , Embrião de Galinha , Fibroblastos/citologia , Fibroblastos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Células HeLa , Humanos , Lisina/genética , Lisina/metabolismo , Fatores de Transcrição Maf Maior/metabolismo , Fator de Transcrição MafB/genética , Fator de Transcrição MafB/metabolismo , Camundongos , Mutação , Células NIH 3T3 , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/genética , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo , Transfecção , delta-Cristalinas/genéticaRESUMO
Cicadas tend to be affected by vicariance reflecting poor mobility of nymphs underground and weak flying ability of adults. However, modern collection records of invasive cicada, combined with records of typhoon tracks, and newly obtained phylogeographic data suggest long distance, relatively instantaneous, dispersal of some vicariantly speciated cicadas. We address the importance of this typhoon dispersal mechanism applied to representative species of east Asian endemic cicadas of Cryptotympana, Mogannia, Euterpnosia and Meimuna. We combine BEAST-dated phylogenic and haplotype network analyses, modern collection data of non-native cicadas available in reports of the Japanese insect associations, modern typhoon records by Japan Meteorological Agency, and our own Quaternary geological constriction data. In conclusion, although Ryukyu endemic cicadas were vicariantly speciated, endemic cicadas on some islands were accidentally dispersed long distances to another island possibly by typhoons, particularly those associated with super typhoons generated since 1.55 Ma.
Assuntos
Distribuição Animal , Tempestades Ciclônicas , Hemípteros , Ilhas , Filogeografia , Animais , DNA Mitocondrial , Haplótipos , Hemípteros/genéticaRESUMO
A self-consistent simulation of secondary electron (SE) emission and charging of a SiO(2) layer with the thickness of several tens of nanometres on Si is incorporated into a trajectory simulation of emitted SEs above the surface, the centre area of which is charged by electron beams (EBs) at the energy range from 300 to 2000 eV. In order to study the influence of the charging of an insulating layer on defect inspection, a pseudo-image is reconstructed from net SE yields calculated at each point of the SiO(2) surface locally applied the positive voltage. The image contrast between charged and uncharged areas is compared with the observation of thermally oxidized layer with the thickness of 24-106 nm on a Si wafer. The image contrast is very sensitive to the thickness of the SiO(2) layer, which is verified by both observed and calculated images. The calculated changes of the images with the layer thickness and the primary electron energy reproduce the experimental observations fairly well. This confirms a highly sensitive detection mechanism for tiny defects in insulating patterns on a metal hard mask for an EB defect inspection equipment.
RESUMO
UNLABELLED: Postnasal drip is believed to be one of the main sources of serious respiratory diseases, such as sinobronchial syndrome. However, there is little direct evidence showing that postnasal drip flows into the trachea and results in the development of inflammatory responses in the lower airway. In the present study, whether postnasal drip entered the respiratory organs and whether material in the trachea moved toward the lungs and the digestive organs were examined by using an experimental model with mice. MATERIALS AND METHODS: In the first set of experiments, 1.0 microL of 51Cr-labeled pseudo-postnasal drip in a normal saline or a glycerin solution was instilled into the nasal cavity of male ICR mice anesthetized with sodium barbital by intraperitoneal injection. In the second set of experiments, the destination of 51Cr-labeled red blood cells (RBCs) after intratracheal instillation was examined in the anesthetized mice. The lungs, the stomach and the intestines were removed from mice killed under anesthesia at various intervals after instillation, and measured for radioactivity. RESULTS: When glycerin solution containing 51Cr (but not normal saline) was instilled in mice, the presence of much higher levels of 51Cr was observed in the lungs. Although the presence of high levels of 51Cr-labeled RBCs was observed in the lungs one hour after instillation radioactivity in the lungs gradually decreased as time went by. On the other hand, radioactivity in the digestive organs gradually increased and peaked three hours after instillation with 51Cr-labeled RBC. CONCLUSION: These results suggest that thicker viscous postnasal drip can flow into the respiratory organs when the host is asleep. In addition, postnasal drip which flows into the trachea can move gradually to the oral side by mucociliary transportation of the tracheal mucosa and thus be swallowed.
Assuntos
Mucosa Nasal/metabolismo , Mucosa Nasal/patologia , Sistema Respiratório/patologia , Anestésicos/farmacologia , Animais , Radioisótopos de Cromo/metabolismo , Eritrócitos/metabolismo , Glicerol/metabolismo , Inflamação , Pulmão/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Sistema Respiratório/imunologia , Rinite/patologia , Sinusite/patologia , SíndromeRESUMO
There is established concept that dendritic cells (DCs) play essential roles in the development of allergic immune responses. However, the influence of H(1) receptor antagonists on DC functions is not well defined. The aim of the present study was to examine the effect of epinastine hydrochloride (EP), the most notable histamine H(1) receptor antagonists in Japan, on Dermatophagoides farinae (Der f)-pulsed mouse bone marrow-derived DCs in vitro and in vivo. EP at more than 25 ng/mL could significantly inhibit the production of IL-6, TNF-alpha and IL-10 from Der f-pulsed DCs, which was increased by Der f challenge in vitro. On the other hand, EP increased the ability of Der f-pulsed DCs to produce IL-12. Intranasal instillation of Der f-pulsed DCs resulted in nasal eosinophilia associated with a significant increase in IL-5 levels in nasal lavage fluids. Der f-pulsed and EP-treated DCs significantly inhibited nasal eosinophila and reduced IL-5. These results indicate that EP inhibits the development of Th2 immune responses through the modulation of DC functions and results in favorable modification of clinical status of allergic diseases.
Assuntos
Alérgenos/imunologia , Células da Medula Óssea/citologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Dermatophagoides farinae/imunologia , Dibenzazepinas/farmacologia , Antagonistas dos Receptores Histamínicos H1/farmacologia , Imidazóis/farmacologia , Administração Intranasal , Animais , Diferenciação Celular , Células Cultivadas , Citocinas/antagonistas & inibidores , Citocinas/biossíntese , Células Dendríticas/citologia , Células Dendríticas/metabolismo , Dibenzazepinas/administração & dosagem , Relação Dose-Resposta a Droga , Feminino , Antagonistas dos Receptores Histamínicos H1/administração & dosagem , Imidazóis/administração & dosagem , Técnicas In Vitro , Inflamação/patologia , Camundongos , Camundongos Endogâmicos BALB C , Líquido da Lavagem Nasal/citologia , Células Th2/efeitos dos fármacos , Células Th2/imunologiaRESUMO
Familial exudative vitreoretinopathy (FEVR) is a human disease characterized by defective retinal angiogenesis and associated complications that can result in vision loss. Defective Wnt/ß-catenin signaling is an established cause of FEVR, whereas other molecular alterations contributing to the disease remain insufficiently understood. Here, we show that integrin-linked kinase (ILK), a mediator of cell-matrix interactions, is indispensable for retinal angiogenesis. Inactivation of the murine Ilk gene in postnatal endothelial cells results in sprouting defects, reduced endothelial proliferation and disruption of the blood-retina barrier, resembling phenotypes seen in established mouse models of FEVR. Retinal vascularization defects are phenocopied by inducible inactivation of the gene for α-parvin (Parva), an interactor of ILK. Screening genomic DNA samples from exudative vitreoretinopathy patients identifies three distinct mutations in human ILK, which compromise the function of the gene product in vitro. Together, our data suggest that defective cell-matrix interactions are linked to Wnt signaling and FEVR.
Assuntos
Barreira Hematorretiniana/metabolismo , Células Endoteliais/metabolismo , Vitreorretinopatias Exsudativas Familiares/genética , Neovascularização Fisiológica/genética , Proteínas Serina-Treonina Quinases/genética , Vasos Retinianos/crescimento & desenvolvimento , Animais , Células Endoteliais/citologia , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Camundongos , Proteínas dos Microfilamentos/genética , Fenótipo , Via de Sinalização Wnt/genéticaRESUMO
The influence of a histamine H1 receptor antagonist, epinastine hydrochloride (EP), on eosinophil functions was examined in vitro and in vivo. The first set of experiments was undertaken to examine whether EP could suppress eosinophilia and IgE hyperproduction induced by Mesocestoides cortii infection in BALB/c mice. The number of peripheral blood eosinophils and levels of IgE were examined 21 days after infection. Oral administration of EP at a daily dose of 0.3 mg/kg, which is the recommended human therapeutic dose, for 21 days was not able to suppress either peripheral blood eosinophilia or IgE hyperproduction, which was observed in mice infected with M. cortii. The second part of the experiment was designed to examine the influence of EP on eosinophil activation induced by stem cell factor (SCF) stimulation in vitro. Eosinophils were obtained from M. cortii-infected mice and stimulated with SCF in the presence of different concentrations of EP for 24 h. The addition of EP into cell cultures suppressed eosinophil activation induced by SCF stimulation as assessed by measuring the contents of acronym for Regulated upon Activation, Normal T cell Expressed and presumably Secreted (RANTES), macrophage inflammatory protein-1beta (MIP-1beta) and leukotriene C4 (LTC4) levels in culture supernatants. The minimum concentration of EP which caused significant suppression of factor productions was 25 ng/ml, which is similar to the concentration in plasma after oral administration of the therapeutic dose in humans. These results may suggest that EP exerts inhibitory effects on eosinophil activation and results in favorable modification of the clinical status of allergic patients.
Assuntos
Dibenzazepinas/farmacologia , Eosinófilos/efeitos dos fármacos , Antagonistas dos Receptores Histamínicos H1/farmacologia , Imidazóis/farmacologia , Inibidores da Agregação Plaquetária/farmacologia , Animais , Células Cultivadas , Infecções por Cestoides/tratamento farmacológico , Infecções por Cestoides/imunologia , Infecções por Cestoides/metabolismo , Quimiocina CCL4/metabolismo , Quimiocina CCL5/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Eosinofilia/sangue , Eosinófilos/imunologia , Imunoglobulina E/metabolismo , Leucotrieno C4/metabolismo , Masculino , Mesocestoides/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Organismos Livres de Patógenos Específicos , Fator de Células-Tronco/farmacologiaRESUMO
The influence of epinastine hydrochloride (EP) on eosinophil survival was examined by an in vitro cell culture technique. Nasal epithelial cells (NECs) were stimulated with 25 ng/ml TNF-alpha in the presence of EP (10 to 30 ng/ml). After 24 h, the culture supernatants were obtained and used as conditioned media of NECs (CM). Eosinophils (1 x 10(3) cells/ml) prepared from healthy human peripheral blood were incubated with 25% CM and eosinophil survival was assessed by trypan blue dye exclusion test 48 h later. CM prepared from NEC cultures pre-treated with TNF-alpha and EP caused a decrease in eosinophil survival as compared with that from NEC cells pre-treated with TNF-alpha alone. The minimum concentration of EP that caused a significant decrease in eosinophil survival was 25 ng/ml. The addition of EP into eosinophil cultures did not cause inhibition of eosinophil survival, which was prolonged by stimulation with granulocyte-macrophage colony-stimulating factor (GM-CSF), even when 40 ng/ml EP was added to cell cultures. We then examined the levels of GM-CSF in CM by ELISA. Treatment of NECs with EP at more than 25 ng/ml, reduced the ability of NECs to produce GM-CSF in response to TNF-alpha stimulation. These results may suggest that EP suppresses eosinophil survival through the suppression of GM-CSF production from NECs induced by inflammatory stimulation and that this suppressive effect contributes, in part, to the therapeutic mode of action of EP on allergic diseases.
Assuntos
Sobrevivência Celular/efeitos dos fármacos , Dibenzazepinas/farmacologia , Eosinófilos/citologia , Antagonistas dos Receptores Histamínicos H1/farmacologia , Imidazóis/farmacologia , Adulto , Técnicas de Cultura de Células , Eosinófilos/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa Nasal/citologia , Mucosa Nasal/patologia , Pólipos Nasais/patologia , Pólipos Nasais/cirurgia , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
The aim of this study was to examine the effect of fexofenadine hydrochloride (FEX), a histamine H1-receptor antagonist, on nitric oxide (NO) production in-vitro and in-vivo. Nasal fibroblasts (5 x 10(5) cells per mL) were stimulated with 25 ng mL(-1) tumour necrosis factor-alpha in the presence of various concentrations of FEX. NO levels in 24-h-culture supernatants were measured by the Griess method and levels of inducible nitric oxide synthase (iNOS) mRNA levels in 12-h-cultured cells were measured by ELISA. FEX at more than 0.5 microg mL(-1) suppressed NO production from fibroblasts by inhibiting expression of iNOS mRNA. We also examined whether FEX could suppress NO production induced by lipopolysaccharide (LPS) stimulation in-vivo. BALB/c mice were treated with 5.0 mg kg(-1) LPS i.p. after daily oral doses of FEX, 1.0 mg kg(-1), for 1-3 weeks. Plasma was obtained 6 h later and NO levels measured by the Griess method. Expression of iNOS mRNA in lung tissues was measured by ELISA 6 h after LPS injection. Oral administration of FEX for 2 and 3 weeks, but not 1 week, significantly suppressed NO levels in plasma through the inhibition of iNOS mRNA expression, which were enhanced by LPS stimulation. These results suggest that the attenuating effect of FEX on NO production may be of therapeutic benefit in allergic diseases.
Assuntos
Antagonistas não Sedativos dos Receptores H1 da Histamina/farmacologia , Óxido Nítrico Sintase Tipo II/efeitos dos fármacos , Óxido Nítrico/biossíntese , Terfenadina/análogos & derivados , Administração Oral , Animais , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Regulação da Expressão Gênica , Antagonistas não Sedativos dos Receptores H1 da Histamina/administração & dosagem , Hipersensibilidade/tratamento farmacológico , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pólipos Nasais/metabolismo , Óxido Nítrico Sintase Tipo II/genética , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Terfenadina/administração & dosagem , Terfenadina/farmacologia , Fatores de Tempo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Tranilast is an anti-allergic agent that blocks the release of chemical mediators, such as histamine and leukotrienes from mast cells, and has been reported to suppress keloid and hypertrophic scar formation. Since matrix metalloproteinases (MMPs) play an essential role in tissue remodelling, this study was undertaken to determine whether tranilast suppresses MMP production from neutrophils after lipopolysaccharide (LPS) stimulation in-vitro. Neutrophils from five healthy donors (1 x 10(5) cells/mL) were stimulated with 1.0 microg mL(-1) LPS in the presence or absence of various concentrations of tranilast for 24 h. MMP-7, MMP-8, MMP-9 and tissue inhibitor of metalloproteinase (TIMP)-1 levels in the culture supernatants were assayed by ELISA. In addition, the influence of tranilast on MMP mRNA expression and transcriptional factor activation in cells cultured for 12 h and 4 h was also evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Tranilast inhibited MMP and TIMP-1 production from neutrophils when cells were treated with the agent at more than 5.0 x 10(-5) M. It also suppressed MMP mRNA expression and transcriptional factor activation induced in neutrophils by LPS stimulation. The results suggest that tranilast inhibits the formation of keloid scarring through the suppression of factors such as MMPs and TIMP, which are essential for tissue remodelling, from inflammatory cells.
Assuntos
Neutrófilos/efeitos dos fármacos , ortoaminobenzoatos/farmacologia , Adulto , Antialérgicos/farmacologia , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Lipopolissacarídeos , Masculino , Metaloproteinase 7 da Matriz/genética , Metaloproteinase 7 da Matriz/metabolismo , Metaloproteinase 8 da Matriz/genética , Metaloproteinase 8 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Pessoa de Meia-Idade , NF-kappa B/metabolismo , Neutrófilos/metabolismo , Neutrófilos/fisiologia , RNA Mensageiro/metabolismo , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismoRESUMO
The effects of intranasal corticosteroids on matrix metalloproteinases (MMPs) and their inhibitors (tissue inhibitors of metalloproteinases (TIMPs)) in the nasal mucosa of patients with allergic rhinitis (AR) are not known. Nasal mucosa biopsy specimens were obtained from AR patients, with or without the administration of fluticasone propionate (FP) nose drops, and from healthy volunteers as controls. The specimens were analyzed by immuno-histochemistry and ELISA for MMP-2, MMP-9, TIMP-1 and TIMP-2. The MMP-9 levels in nasal mucosa extracts in the AR patients were significantly higher than in the controls. A significant suppressive effect of FP on the MMP-9 levels was shown. The control subjects showed no MMP- or TIMP-positive cells, whereas such positive cells were clearly present in the AR patients. No MMP- or TIMP-positive cells were detected after topical application of FP. These results suggest that the suppressive effect of FP on MMP expression is, in part, responsible for its clinical efficacy in AR.
Assuntos
Androstadienos/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Inibidores de Metaloproteinases de Matriz , Mucosa Nasal/metabolismo , Rinite Alérgica Perene/tratamento farmacológico , Administração Tópica , Adolescente , Adulto , Androstadienos/administração & dosagem , Anti-Inflamatórios/administração & dosagem , Biópsia , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Feminino , Fluticasona , Humanos , Imuno-Histoquímica , Masculino , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , Metaloproteinases da Matriz/biossíntese , Metaloproteinases da Matriz/genética , Pessoa de Meia-Idade , Mucosa Nasal/efeitos dos fármacos , Rinite Alérgica Perene/metabolismo , Inibidor Tecidual de Metaloproteinase-1/análise , Inibidor Tecidual de Metaloproteinase-2/análiseRESUMO
Thymus- and activation-regulated chemokine (TARC) is an important molecule in the development and maintenance of allergic diseases. However, there is little information about the influence of anti-allergic agents on TARC production. The aim of this study is to examine the influence of epinastine hydrochloride, an H1-receptor antagonist, on TARC production from human peripheral blood CD4+ T cells using an in-vitro cell culture technique. CD4+ T cells prepared from healthy subjects were cultured in wells coated with a combination of OKT3 and anti-CD28 monoclonal antibody in the presence or absence of epinastine HCl for 24 h. The cells were also stimulated with interleukin (IL)-4 in a similar manner. Levels of TARC and IL-4 in culture supernatants were examined by ELISA. The addition of epinastine HCl exerted a dose-dependent suppressive effect on the production of both TARC and IL-4 from CD4+ T cells under co-stimulatory molecule stimulation. The minimum concentration of the agent showing a significant suppressive effect on TARC and IL-4 production was 5.0 microM and 2.5 microM, respectively. Epinastine HCl also suppressed the ability of cells to produce TARC in response to IL-4 stimulation, when the agent was added to cell cultures at more than 2.5 microM. It was concluded that this inhibitory action of epinastine HCl may be partially responsible for epinastine's attenuating effect on allergic diseases.
Assuntos
Linfócitos T CD4-Positivos/efeitos dos fármacos , Quimiocinas CC/antagonistas & inibidores , Dibenzazepinas/farmacologia , Antagonistas dos Receptores Histamínicos H1/farmacologia , Imidazóis/farmacologia , Subpopulações de Linfócitos T/efeitos dos fármacos , Adulto , Linfócitos T CD4-Positivos/imunologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Quimiocina CCL17 , Quimiocinas CC/biossíntese , Quimiocinas CC/genética , Relação Dose-Resposta Imunológica , Humanos , Separação Imunomagnética , Interleucina-4/antagonistas & inibidores , Interleucina-4/imunologia , Ativação Linfocitária/efeitos dos fármacos , Pessoa de Meia-Idade , Muromonab-CD3 , NF-kappa B/antagonistas & inibidores , NF-kappa B/imunologia , Subpopulações de Linfócitos T/imunologiaRESUMO
Angiogenic blood vessel growth requires several distinct but integrated cellular activities. Endothelial cell sprouting and proliferation lead to the expansion of the vasculature and give rise to a highly branched, immature plexus, which is subsequently reorganized into a mature and stable network. Although it is known that integrin-mediated cell-matrix interactions are indispensable for embryonic angiogenesis, little is known about the function of integrins in different steps of vascular morphogenesis. Here, by investigating the integrin ß1-subunit with inducible and endothelial-specific gene targeting in the postnatal mouse retina, we show that ß1 integrin promotes endothelial sprouting but is a negative regulator of proliferation. In maturing vessels, integrin ß1 is indispensable for proper localization of VE-cadherin and thereby cell-cell junction integrity. The sum of our findings establishes that integrin ß1 has critical functions in the growing and maturing vasculature, and is required for the formation of stable, non-leaky blood vessels.
Assuntos
Antígenos CD/metabolismo , Caderinas/metabolismo , Endotélio/crescimento & desenvolvimento , Integrina beta1/metabolismo , Junções Intercelulares/fisiologia , Morfogênese/fisiologia , Neovascularização Fisiológica/fisiologia , Vasos Retinianos/crescimento & desenvolvimento , Animais , Western Blotting , Encéfalo/anatomia & histologia , Proliferação de Células , Endotélio/metabolismo , Marcação de Genes , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Imunoprecipitação , Junções Intercelulares/metabolismo , Camundongos , Microscopia Eletrônica , RNA Interferente Pequeno/genética , Reação em Cadeia da Polimerase em Tempo Real , Vasos Retinianos/ultraestruturaRESUMO
The influence of fluticasone propionate (FP) on vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) production from nasal polyp fibroblasts (NPFs) was examined in vitro. NPFs, at a density of 2.5 x 10(5) cells/ml, were stimulated with 25 ng/ml TNF-alpha in the presence or absence of FP for 24 h. FP at more than 10(-6) M could suppress VEGF production from NPFs. bFGF production induced by TNF-alpha stimulation was also suppressed by FP, when the agent was added to cell cultures at more than 10(-7) M. The present results also showed the suppressive activity of FP on mRNA expression for VEGF and bFGF, which were increased by TNF-alpha stimulation.
Assuntos
Androstadienos/farmacologia , Anti-Inflamatórios/farmacologia , Fator 2 de Crescimento de Fibroblastos/metabolismo , Fibroblastos/efeitos dos fármacos , Mucosa Nasal/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Células Cultivadas , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Fator 2 de Crescimento de Fibroblastos/genética , Fibroblastos/metabolismo , Fluticasona , Expressão Gênica , Humanos , Mucosa Nasal/metabolismo , Mucosa Nasal/patologia , Pólipos Nasais/patologia , RNA Mensageiro/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
OBJECTIVE: To examine the possible mechanisms underlying the therapeutic mode of action of glucocorticoids (GCs) in nasal polyposis. MATERIAL AND METHODS: The effects of GCs on nasal polyps were firstly evaluated by examining the growth of fibroblasts derived from 10 nasal polyps in vitro. Subsequently, the ability of GCs to induce apoptotic cell death in fibroblasts was examined. RESULTS: Addition of betamethasone 21-phosphate (BET) at a concentration of > 1 x 10(-3) M to cell cultures inhibited cell growth in all cases examined. BET and dexamethasone 21-phosphate, but not testosterone or estradiol, caused apoptotic cell death in 2/10 nasal polyp fibroblasts, as assessed by agarose gel electrophoresis, when the cells were cultured with the agents for > 96 h. The minimum concentration of agent needed to cause apoptosis was 1 x 10(-3) M, which is half of the recommended therapeutic dose. CONCLUSION: The present findings suggest that topical application of GCs in nasal polyposis patients suppresses proliferation of fibroblasts in polyps and results in favorable modification of the clinical status of these patients.
Assuntos
Anti-Inflamatórios/farmacologia , Apoptose/efeitos dos fármacos , Betametasona/análogos & derivados , Dexametasona/análogos & derivados , Fibroblastos/efeitos dos fármacos , Glucocorticoides/farmacologia , Pólipos Nasais/tratamento farmacológico , Adulto , Anti-Inflamatórios/administração & dosagem , Apoptose/genética , Betametasona/farmacologia , Células Cultivadas , Dexametasona/farmacologia , Feminino , Fibroblastos/citologia , Fibroblastos/metabolismo , Glucocorticoides/administração & dosagem , Humanos , Masculino , Pessoa de Meia-Idade , Pólipos Nasais/patologiaRESUMO
OBJECTIVE: To examine the influence of fluticasone propionate (FP) on matrix metalloproteinase (MMP) production from nasal polyp fibroblasts in vitro. MATERIAL AND METHODS: Fibroblasts derived from five nasal polyps were stimulated with tumor necrosis factor (TNF)-alpha in the presence of various concentrations of FP. The influence of FP on MMP production was assessed by examining the levels of MMP-2 and -9 in culture supernatants using ELISA. We also examined the influence of FP on MMP mRNA expression using reverse transcriptase polymerase chain reaction. RESULTS: The addition of FP caused significant suppression of MMP-2 and -9 production from nasal polyp fibroblasts in response to TNF-alpha stimulation. MMP mRNA expression was also suppressed by the addition of FP to cell cultures. The minimum concentration of the agent required to cause suppression was 10(-5) M. CONCLUSION: These results suggest that the inhibitory action of FP on tissue remodeling may underlie the clinical efficacy of corticosteroids in nasal polyposis.
Assuntos
Androstadienos/farmacologia , Anti-Inflamatórios/farmacologia , Fibroblastos/efeitos dos fármacos , Metaloproteinases da Matriz/metabolismo , Pólipos Nasais/patologia , Adulto , Androstadienos/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Células Cultivadas , Feminino , Fibroblastos/enzimologia , Fluticasona , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Inibidores de Metaloproteinases de Matriz , Metaloproteinases da Matriz/genética , Pessoa de Meia-Idade , Pólipos Nasais/tratamento farmacológico , Pólipos Nasais/enzimologia , RNA Mensageiro/metabolismo , Inibidores Teciduais de Metaloproteinases/antagonistas & inibidores , Inibidores Teciduais de Metaloproteinases/genética , Inibidores Teciduais de Metaloproteinases/metabolismoRESUMO
Eosinophils are well known to play essential roles in the development and maintenance of allergic diseases. However, the influence of histamine H1 receptor antagonists on eosinophil functions, especially chemokine production, are not well-defined. Therefore, in the present study, we examined the influence of histamine H1 receptor antagonist on chemokine production by eosinophils through the use of levocetirizine in vitro and in vivo. Eosinophils prepared from mice were stimulated with specific antigens in the presence of different concentrations of levocetirizine. After 24 h, regulated on activation normal T cell expressed and secreted (RANTES) and eotaxin levels in culture supernatants were measured by enzyme-linked immunosorbent assay (ELISA). Patients with Japanese cedar pollinosis were treated with 5 mg levocetirizine once a day for four weeks during the pollen season (February 2012 to April 2012). RANTES and eotaxin levels in nasal secretions were also examined by ELISA. The addition of levocetirizine to eosinophil cultures caused a dose-dependent decrease in the ability of cells to produce RANTES and eotaxin in response to antigen stimulation, and the minimum concentration that caused a significant decrease was 0.05 µM. Although cetirizine also exerted suppressive effects on the production of RANTES and eotaxin by eosinophils, the minimum concentration that caused significant suppression was 0.15 µM, which was three-times higher than that of levocetirizine. Oral administration of levocetirizine for four weeks also reduced RANTES and eotaxin levels in nasal secretions from patients with pollinosis, along with attenuation of clinical symptoms. The ability of levocetirizine to reduce RANTES and eotaxin levels may account, at least in part, for the clinical efficacy of the agent for allergic disorders, including allergic rhinitis.
Assuntos
Cetirizina/farmacologia , Quimiocina CCL5/biossíntese , Fatores Quimiotáticos/biossíntese , Eosinófilos/efeitos dos fármacos , Eosinófilos/metabolismo , Adulto , Idoso , Animais , Antígenos/imunologia , Estudos de Casos e Controles , Eosinófilos/imunologia , Feminino , Expressão Gênica , Antagonistas não Sedativos dos Receptores H1 da Histamina/farmacologia , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Contagem de Leucócitos , Masculino , Camundongos , Pessoa de Meia-Idade , RNA Mensageiro/genética , Rinite Alérgica Sazonal/diagnóstico , Rinite Alérgica Sazonal/tratamento farmacológico , Rinite Alérgica Sazonal/genética , Rinite Alérgica Sazonal/imunologia , Rinite Alérgica Sazonal/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
OBJECTIVES: Osteopontin (OPN), a multifunctional glycoprotein secreted from a wide variety of cells after inflammatory stimulation, is well accepted to contribute to the development of allergic diseases. However, the influence of histamine H1 receptor antagonists (antihistamines) on OPN functions is not well understood. The present study was undertaken to examine the influence of antihistamines on OPN functions in vitro. METHODS: Human nasal epithelial cells (5 × 10(5) cells) were stimulated with 250 ng/mL OPN in the presence of either desloratadine (DL), fexofenadine (FEX), or levocetirizine (LCT). The levels of OPN, GM-CSF, Eotaxin, and RANTES in 24 h culture supernatants were examined by ELISA. The influence of LCT on mRNA expression and transcription factor activation in cells were also examined by real-time RT-PCR and ELISA, respectively. KEY FINDINGS: The antihistamines examined significantly suppressed the production of GM-CSF, Eotaxin, and RANTES from cells after OPN stimulation. LCT also exhibited the suppression of mRNA expression for chemokines and transcription factor, NF- κ B and AP-1, activation, which were increased by the stimulation of cells with OPN. CONCLUSIONS: The suppressive activity of LCT on OPN functions on nasal epithelial cells may be responsible for the attenuating effect of the agent on allergic diseases.