RESUMO
A rechargeable acidic polymer-air battery was firstly fabricated with poly(2,5-dihydroxy-1,4-benzoquinone-3,6-methylene) (PDBM) as the anode, the conventional Pt/C cathode catalyst, and acidic aqueous electrolyte (pH 1). This battery yielded a high discharging capacity of 349 mA h gpolymer-1 with a long-lifetime of >500 cycles and high rate capabilities (up to 10C).
RESUMO
Human immunodeficiency virus type 1 (HIV-1) is a major causative agent of acquired immune deficiency syndrome. Subtype C (HIV-1C) is the most prevalent HIV-1 subtype worldwide. Although it is highly prevalent in Nepal, genotypic information on Nepalese HIV-1C is limited. We herein investigated the origin and dynamics of HIV-1C in Nepal. Nearly full-length sequencing of Nepalese HIV-1C strains and phylogenetic analyses were performed. The results obtained showed that Nepalese HIV-1C is closely related to the Indian and southern African strains and the introduction of HIV-1C into Nepal was estimated to be in the mid-1980s. These results suggest that multiple HIV-1C strains entered Nepal in the mid-1980s, and this was followed by a marked increase in the number of infection cases for the next decade. These results reflect the current transmission dynamics of HIV-1C strains in Nepal and provide valuable information for HIV monitoring and vaccine development.
Assuntos
Genótipo , HIV-1/classificação , HIV-1/genética , Infecções por HIV/epidemiologia , Infecções por HIV/virologia , Humanos , Nepal/epidemiologia , Filogenia , Análise de Sequência de DNARESUMO
Respiratory syncytial virus (RSV) is one of the most prevalent causative agents of lower respiratory tract infections worldwide, especially in infants around 3 to 4months old. Infants at such a young age have maternally-transferred passive antibodies against RSV but do not have active immune systems efficient enough for the control of RSV infection. In order to elucidate age-specific profiles of immune responses against RSV protection, antibody responses were examined by using blood samples in both acute and convalescent phases obtained from child patients and adult patients. In addition to the serum neutralization activity, antibody responses to the RSV fusion protein (F protein) were dissected by analyzing levels of total IgG, IgG subclasses, the binding stability, and the levels of antibody for the neutralization epitopes. It was suggested that children's antibody responses against RSV are matured over months and years in at least 5 stages based on 1) levels of the neutralization titer and IgG3 for F protein in the convalescent phase, 2) geometric mean ratios of the neutralization titers and levels of IgG1 and IgG2 for F protein in the convalescent phase compared to those levels in the acute phase, 3) the affinity maturation of IgG for F protein and the cross reactivity of IgG for RSV glycoproteins of groups A and B, 4) levels of neutralization epitope-specific IgG, and 5) augmentation of overall antibody responses due to repetitive RSV infection.
Assuntos
Anticorpos Antivirais/imunologia , Infecções por Vírus Respiratório Sincicial/imunologia , Vírus Sincicial Respiratório Humano/imunologia , Proteínas Virais de Fusão/imunologia , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antivirais/sangue , Linhagem Celular Tumoral , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Interações Hospedeiro-Patógeno/imunologia , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Infecções por Vírus Respiratório Sincicial/sangue , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sincicial Respiratório Humano/fisiologiaRESUMO
BACKGROUND: It has been clinically demonstrated that intradermal (ID) vaccines have a potential to confer a superior immunogenic profile compared to intramuscular (IM) or subcutaneous (SC) vaccines. In terms of distribution of a vaccine antigen depending on the administration routes, at least two independent immunogenic pathways of the vaccines have been proposed: (1) the antigen recognition by the immune cells present at the vaccine-administered site and (2) the antigen recognition by the lymph node (LN)-resident immune cells through the lymphatic flow from the vaccine-administered site after the antigen is directly delivered into the draining LNs. OBJECTIVE: In order to clarify the key components for the immunogenic pathway of the ID vaccine, the correlation between the kinetics of the antigen distribution to the draining LNs and antibody responses to the antigen were evaluated. METHODS: We compared the antibody responses in the groups with by surgical removal of the administration site immediately after the ID administration, and by surgical removal of the draining LNs before the ID administration. RESULTS: The results suggested that the efficient and direct antigen delivery to the draining LNs plays an important role in the antibody responses to the ID vaccine. Indeed, it was confirmed that the direct administration into the draining LNs with the antigen elicited comparable levels of the antibody responses with the ID vaccine. At the cellular level, it was shown that the LN-resident immune cells such as B cells, dendritic cells, and macrophages including medullary macrophages and subcapsular sinus macrophages interacting with the antigens following the ID administration. Finally, we demonstrated by immunofluorescence analysis that the lymphatic vessels are more diffusely distributed in the dermis as compared with the subcutaneous area and muscle. CONCLUSION: The results of the present study suggested that the skin is an optimal tissue to facilitate the vaccine antigen access to the draining LNs, which is an important immunogenic pathway of the ID vaccine. Further elucidation of regulatory mechanisms underlying such an immunogenic pathway of the ID vaccine would provide us with elements for the development of novel adjuvants and devices to enhance the immunogenicity of the ID vaccines.
Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/administração & dosagem , Imunogenicidade da Vacina , Vacinas contra Influenza/administração & dosagem , Linfonodos/imunologia , Pele/imunologia , Vacinação , Animais , Anticorpos Antivirais/sangue , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vacinas contra Influenza/imunologia , Injeções Intradérmicas , Injeções Subcutâneas , Cinética , Linfonodos/metabolismo , Camundongos Endogâmicos BALB C , Pele/metabolismoRESUMO
It is thought that apolipoprotein A-I (apoA-I) spontaneously exchanges between high-density lipoprotein (HDL)-bound and lipid-free states, which is relevant to the occurrence of preß-HDL particles in plasma. To improve our understanding of the mechanistic basis for this phenomenon, we performed kinetic and thermodynamic analyses for apoA-I exchange between discoidal HDL-bound and lipid-free forms using fluorescence-labeled apoA-I variants. Gel filtration experiments demonstrated that addition of excess lipid-free apoA-I to discoidal HDL particles promotes exchange of apoA-I between HDL-associated and lipid-free pools without alteration of the steady-state HDL particle size. Kinetic analysis of time-dependent changes in NBD fluorescence upon the transition of NBD-labeled apoA-I from HDL-bound to lipid-free state indicates that the exchange kinetics are independent of the collision frequency between HDL-bound and lipid-free apoA-I, in which the lipid binding ability of apoA-I affects the rate of association of lipid-free apoA-I with the HDL particles and not the rate of dissociation of HDL-bound apoA-I. Thus, C-terminal truncations or mutations that reduce the lipid binding affinity of apoA-I strongly impair the transition of lipid-free apoA-I to the HDL-bound state. Thermodynamic analysis of the exchange kinetics demonstrated that the apoA-I exchange process is enthalpically unfavorable but entropically favorable. These results explain the thermodynamic basis of the spontaneous exchange reaction of apoA-I associated with HDL particles. The altered exchangeability of dysfunctional apoA-I would affect HDL particle rearrangement, leading to perturbed HDL metabolism.
Assuntos
Apolipoproteína A-I/metabolismo , Lipoproteínas HDL/metabolismo , Termodinâmica , Apolipoproteína A-I/farmacocinética , Cinética , Lipoproteínas HDL/farmacocinética , Ligação Proteica/fisiologia , Engenharia de Proteínas/métodosRESUMO
BACKGROUND: Rapid desensitization transiently prevents severe allergic reactions, allowing administration of life-saving therapies in previously sensitized patients. However, the mechanisms underlying successful rapid desensitization are not fully understood. OBJECTIVES: We sought to investigate whether the mast cell (MC) is an important target of rapid desensitization in mice sensitized to exhibit IgE-dependent passive systemic anaphylaxis in vivo and to investigate the antigen specificity and underlying mechanisms of rapid desensitization in our mouse model. METHODS: C57BL/6 mice (in vivo) or primary isolated C57BL/6 mouse peritoneal mast cells (PMCs; in vitro) were passively sensitized with antigen-specific anti-2,4-dinitrophenyl IgE, anti-ovalbumin IgE, or both. MCs were exposed over a short period of time to increasing amounts of antigen (2,4-dinitrophenyl-human serum albumin or ovalbumin) in the presence of extracellular calcium in vitro or by means of intravenous administration to sensitized mice in vivo before challenging the mice with or exposing the PMCs to optimal amounts of specific or irrelevant antigen. RESULTS: Rapidly exposing mice or PMCs to progressively increasing amounts of specific antigen inhibited the development of antigen-induced hypothermia in sensitized mice in vivo and inhibited antigen-induced PMC degranulation and prostaglandin D2 synthesis in vitro. Such MC hyporesponsiveness was induced antigen-specifically and was associated with a significant reduction in antigen-specific IgE levels on MC surfaces. CONCLUSIONS: Rapidly exposing MCs to progressively increasing amounts of antigen can both enhance the internalization of antigen-specific IgE on the MC surface and also desensitize these cells in an antigen-specific manner in vivo and in vitro.
Assuntos
Anafilaxia/terapia , Antígenos/imunologia , Dessensibilização Imunológica/métodos , Imunoglobulina E/imunologia , Mastócitos/imunologia , 2,4-Dinitrofenol/imunologia , Animais , Anticorpos Anti-Idiotípicos/imunologia , Especificidade de Anticorpos , Antígenos/administração & dosagem , Antígenos/metabolismo , Humanos , Hipersensibilidade , Camundongos , Camundongos Endogâmicos C57BL , Ovalbumina/imunologia , Fatores de TempoRESUMO
BACKGROUND: Mast cells express receptors for complement anaphylatoxins C3a and C5a (ie, C3a receptor [C3aR] and C5a receptor [C5aR]), and C3a and C5a are generated during various IgE-dependent immediate hypersensitivity reactions in vivo. However, it is not clear to what extent mast cell expression of C3aR or C5aR influences C3a- or C5a-induced cutaneous responses or IgE-dependent mast cell activation and passive cutaneous anaphylaxis (PCA) in vivo. OBJECTIVE: We sought to assess whether mouse skin mast cell expression of C3aR or C5aR influences (1) the cells' responsiveness to intradermal injections of C3a or C5a or (2) the extent of IgE-dependent mast cell degranulation and PCA in vivo. METHODS: We measured the magnitude of cutaneous responses to intradermal injections of C3a or C5a and the extent of IgE-dependent mast cell degranulation and PCA responses in mice containing mast cells that did or did not express C3aR or C5aR. RESULTS: The majority of the skin swelling induced by means of intradermal injection of C3a or C5a required that mast cells at the site expressed C3aR or C5aR, respectively, and the extent of IgE-dependent degranulation of skin mast cells and IgE-dependent PCA was significantly reduced when mast cells lacked either C3aR or C5aR. IgE-dependent PCA responses associated with local increases in C3a levels occurred in antibody-deficient mice but not in mice deficient in FcÉRIγ. CONCLUSION: Expression of C3aR and C5aR by skin mast cells contributes importantly to the ability of C3a and C5a to induce skin swelling and can enhance mast cell degranulation and inflammation during IgE-dependent PCA in vivo.
Assuntos
Imunoglobulina E/metabolismo , Inflamação/imunologia , Mastócitos/imunologia , Receptor da Anafilatoxina C5a/biossíntese , Receptores de Complemento/biossíntese , Pele/imunologia , Anafilatoxinas/genética , Anafilatoxinas/imunologia , Animais , Células Cultivadas , Complemento C3a/genética , Complemento C3a/imunologia , Complemento C3a/metabolismo , Complemento C5a/genética , Complemento C5a/imunologia , Complemento C5a/metabolismo , Feminino , Imunoglobulina E/genética , Inflamação/genética , Inflamação/metabolismo , Inflamação/patologia , Mastócitos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Receptor da Anafilatoxina C5a/genética , Receptor da Anafilatoxina C5a/imunologia , Receptores de Complemento/genética , Receptores de Complemento/imunologia , Receptores de Complemento/metabolismo , Pele/metabolismo , Pele/patologiaRESUMO
Cromolyn, widely characterized as a 'mast cell stabilizer', has been used in mice to investigate the biological roles of mast cells in vivo. However, it is not clear to what extent cromolyn can either limit the function of mouse mast cells or influence biological processes in mice independently of effects on mast cells. We confirmed that cromolyn (at 10 mg/kg in vivo or 10-100 µM in vitro) can inhibit IgE-dependent mast cell activation in rats in vivo (measuring Evans blue extravasation in passive cutaneous anaphylaxis (PCA) and increases in plasma histamine in passive systemic anaphylaxis (PSA)) and in vitro (measuring peritoneal mast cell (PMC) ß-hexosaminidase release and prostaglandin D(2) synthesis). However, under the conditions tested, cromolyn did not inhibit those mast cell-dependent responses in mice. In mice, cromolyn also failed to inhibit the ear swelling or leukocyte infiltration at sites of PCA. Nor did cromolyn inhibit IgE-independent degranulation of mouse PMCs induced by various stimulators in vitro. At 100 mg/kg, a concentration 10 times higher than that which inhibited PSA in rats, cromolyn significantly inhibited the increases in plasma concentrations of mouse mast cell protease-1 (but not of histamine) during PSA, but had no effect on the reduction in body temperature in this setting. Moreover, this concentration of cromolyn (100 mg/kg) also inhibited LPS-induced TNF production in genetically mast cell-deficient C57BL/6-Kit(W-sh/W-sh) mice in vivo. These results question cromolyn's effectiveness and selectivity as an inhibitor of mast cell activation and mediator release in the mouse.
Assuntos
Anafilaxia/tratamento farmacológico , Antiasmáticos/farmacologia , Cromolina Sódica/farmacologia , Mastócitos/efeitos dos fármacos , Análise de Variância , Anafilaxia/metabolismo , Animais , Quimiocina CCL2/metabolismo , Quimases/sangue , Quimases/metabolismo , Cromolina Sódica/uso terapêutico , Relação Dose-Resposta a Droga , Azul Evans , Extravasamento de Materiais Terapêuticos e Diagnósticos , Feminino , Histamina/sangue , Humanos , Imunoglobulina E/metabolismo , Leucócitos/efeitos dos fármacos , Leucócitos/metabolismo , Masculino , Mastócitos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Anafilaxia Cutânea Passiva/efeitos dos fármacos , Peritônio/citologia , Peritônio/efeitos dos fármacos , Ratos , Especificidade da Espécie , Fator de Necrose Tumoral alfa/sangue , beta-N-Acetil-Hexosaminidases/metabolismoRESUMO
Mesoporous polyacrylonitrile (PAN) monolith has been fabricated by a template-free approach using the unique affinity of PAN towards a water/dimethyl sulfoxide (DMSO) mixture. A newly developed Thermally Induced Phase Separation Technique (TIPS) has been used to obtain the polymer monoliths and their microstructures have been controlled by optimizing the concentration and cooling temperature.
RESUMO
We evaluated the pharmacological effect of statins (3-hydroxy-3-methylglutaryl-CoA reductase inhibitors) on mast cell degranulation in RBL-2H3 cells. A hydrophilic statin (pravastatin) did not inhibit degranulation induced by dinitrophenol-human serum albumin (DNP-HSA); in contrast, lipophilic statins (simvastatin, fluvastatin and atorvastatin) inhibited DNP-HSA-induced degranulation in that order. The inhibitory effects were completely attenuated by simultaneous treatment with 100-1000 microM mevalonic acid for 4 h. We used fluvastatin to clarify the mechanism of the statin-mediated inhibitory action of mast cell degranulation. Fluvastatin (3 microM) had no effect on Ca(2+) release from the endoplasmic reticulum or Ca(2+) influx in the DNP-HSA- or thapsigargin-stimulated cells. Fluvastatin treatment also had no effect on the total granule content of the cell or sensitivity to DNP-HSA and IgE. Fluvastatin (3 microM, 24 h treatment) also failed to affect the morphology, proliferation, and viability of RBL-2H3 cells. Geranylgeranyl transferase inhibitor, GGTI-286 (20 microM), but not farnesyl transferase inhibitor, FPTIII (20 microM), inhibited the DNP-HSA-induced degranulation. The GGTI-286-induced inhibitory action was not associated with a decrease in the cytoplasmic Ca(2+) level. In conclusion, fluvastatin at a lower concentration range inhibited DNP-HSA-induced degranulation without affecting the cytoplasmic Ca(2+) response and also without changing the amount of granule content and proliferation of the mast cells. The statin-induced inhibitory action may be mediated by the suppression of geranylgeranyl transferase via the depletion of intracellular mevalonic acid.
Assuntos
Cálcio/metabolismo , Degranulação Celular/efeitos dos fármacos , Citoplasma/efeitos dos fármacos , Ácidos Graxos Monoinsaturados/farmacologia , Indóis/farmacologia , Mastócitos/efeitos dos fármacos , Alquil e Aril Transferases/antagonistas & inibidores , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Colesterol/biossíntese , Citoplasma/metabolismo , Grânulos Citoplasmáticos/efeitos dos fármacos , Grânulos Citoplasmáticos/metabolismo , Dinitrofenóis/farmacologia , Inibidores Enzimáticos/farmacologia , Fluvastatina , Humanos , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Leucina/análogos & derivados , Leucina/farmacologia , Mastócitos/citologia , Mastócitos/metabolismo , Ácido Mevalônico/metabolismo , Organofosfonatos/farmacologia , Fosfatos de Poli-Isoprenil/metabolismo , Albumina Sérica/farmacologia , Tapsigargina/farmacologiaRESUMO
Mast cells are secretory cells that release their granules, which contain inflammatory mediators. Some recent data suggested that cytoskeletons play a role in this process. However, the role of microtubules in Ca2+ signaling has not yet been well defined. In this study, we demonstrate that the microtubule cytoskeleton is important to maintain Ca2+ influx in the degranulation pathway of mast cells, using the microtubule depolymerizers nocodazole and colchicine. The microtubule depolymerizers inhibited Ag-induced degranulation in RBL-2H3 cells and bone marrow-derived mast cells. When the cells were stimulated with Ag in the presence of the microtubule depolymerizers, the Ca2+ influx was decreased without affecting Ca2+ release from the endoplasmic reticulum (ER). Capacitative Ca2+ entry, which was induced by inhibitors of Ca(2+)-ATPase in the ER membrane, thapsigargin and cyclopiazonic acid, was also decreased by nocodazole. Fluorescent probe analysis demonstrated that nocodazole disrupted microtubule formation and changed the cytoplasmic distribution of the ER. The microtubule depolymerizers attenuated the passive cutaneous anaphylaxis reaction in back skin of Sprague Dawley rats. These results suggest that the microtubule cytoskeleton in mast cells is important to maintain Ag-induced capacitative Ca2+ entry, which is responsible for degranulation and the allergic response.
Assuntos
Sinalização do Cálcio/efeitos dos fármacos , Degranulação Celular/efeitos dos fármacos , Mastócitos/efeitos dos fármacos , Mastócitos/fisiologia , Microtúbulos/efeitos dos fármacos , Microtúbulos/fisiologia , Animais , Linhagem Celular , Colchicina/farmacologia , Retículo Endoplasmático/efeitos dos fármacos , Técnicas In Vitro , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microtúbulos/imunologia , Nocodazol/farmacologia , Anafilaxia Cutânea Passiva/efeitos dos fármacos , Ratos , Ratos Sprague-DawleyRESUMO
In the mast cell signaling pathways, the binding of immunoglobulin E (IgE) to FcepsilonRI, its high-affinity receptor, is generally thought to be a passive step. In this study, we examined the effect of IgE alone, that is, without antigen stimulation, on the degranulation in mast cells. Monomeric IgE (500-5,000 ng/ml) alone increased cytosolic Ca2+ level ([Ca2+]i) and induced degranulation in rat basophilic leukemia (RBL)-2H3 mast cells. Monomeric IgE (5,000 ng/ml) alone also increased [Ca2+]i and induced degranulation in bone marrow-derived mast cells. Interestingly, monomeric IgE (5-50 ng/ml) alone, in concentrations too low to induce degranulation, increased filamentous actin content in RBL-2H3 mast cells. We next examined whether actin dynamics affect the IgE alone-induced RBL-2H3 mast cell activation pathways. Cytochalasin D inhibited the ability of IgE alone (50 ng/ml) to induce de novo actin assembly. In cytochalasin D-treated cells, IgE (50 ng/ml) alone increased [Ca2+]i and induced degranulation. We have summarized the current findings into two points. First, IgE alone increases [Ca2+]i and induces degranulation in mast cells. Second, IgE, at concentrations too low to increase either [Ca2+]i or degranulation, significantly induces actin assembly, which serves as a negative feedback control in the mast cell Ca2+ signaling and degranulation.
Assuntos
Actinas/efeitos dos fármacos , Actinas/fisiologia , Sinalização do Cálcio/fisiologia , Degranulação Celular/fisiologia , Imunoglobulina E/farmacologia , Mastócitos/fisiologia , Actinas/metabolismo , Animais , Cálcio/metabolismo , Degranulação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Membranas Intracelulares/metabolismo , Mastócitos/efeitos dos fármacos , Concentração Osmolar , RatosRESUMO
In the isolated rat aorta, a ROCKs (rhoA-dependent coiled coil serine/threonine kinases) inhibitor, Y-27632, inhibited the contractions induced not only by receptor agonists but also by high K(+) with the similar IC(50) values (0.8 - 4.9 microM). However, Y-27632 did not inhibit the increment of cytosolic Ca(2+) concentration ([Ca(2+)](i)) due to these stimulants. The Y-27632-induced inhibition of contraction was accompanied by an inhibition of myocin light chain (MLC) phosphorylation, although inhibition of contraction was stronger than that of MLC phosphorylation during the initial phase of contraction. Y-27632 had no effect on the myocin light chain kinase (MLCK) activity. This inhibitor also did not directly change the phosphatase activity. These results suggest that Y-27632 is a selective inhibitor of ROCKs with no direct inhibitory effect on [Ca(2+)](i), calmodulin, MLCK, or phosphatase. Y-27632 disrupted the actin filament network and decreased the filamentous actin, implying that the stronger inhibition by Y-27632 on early phase of contraction than MLC phosphorylation may be explained by this effect. These results suggest that the high K(+)-induced MLC phosphorylation and contraction are mediated not only by the classical Ca(2+)/calmodulin-dependent MLCK system but also by a novel MLC phosphorylation pathway involving ROCKs. One of the possibilities is that high K(+) activates ROCKs to inhibit myosin phosphatase resulting in an augmentation of MLC phosphorylation and contraction.
Assuntos
Amidas/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/metabolismo , Piridinas/farmacologia , Transdução de Sinais , Actinas/metabolismo , Animais , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/metabolismo , Aorta Torácica/fisiologia , Cálcio/metabolismo , Linhagem Celular , Técnicas In Vitro , Peptídeos e Proteínas de Sinalização Intracelular , Contração Isométrica/efeitos dos fármacos , Masculino , Artérias Mesentéricas/efeitos dos fármacos , Artérias Mesentéricas/metabolismo , Artérias Mesentéricas/fisiologia , Cadeias Leves de Miosina/metabolismo , Quinase de Cadeia Leve de Miosina/metabolismo , Fosfatase de Miosina-de-Cadeia-Leve/metabolismo , Fosforilação , Coelhos , Ratos , Ratos Wistar , Especificidade da Espécie , Quinases Associadas a rhoRESUMO
We examined the effects of KD3-671 (2-propyl-8-oxo-1-[(2'-(H-tetrazole-5-yl)biphenyl-4-yl)methyl]-4,5,6,7-tetrahydrocycloheptimidazole), an angiotensin II type1 receptor antagonist, on an experimental rat model of mesangioproliferative glomerulonephritis, anti-Thy-1 nephritis. Anti-Thy-1 nephritis was induced by intravenous injection of 300 microg/kg of anti-Thy-1.1 monoclonal antibody into rats. KD3-671 (3, 10, 30 mg/kg per day) or enalapril (30 mg/kg per day), an angiotensin II converting enzyme inhibitor, was given p.o. once daily from the day before the antibody injection (the 1st day) to the 15th day after. KD3-671 significantly inhibited an increase in the number of total and proliferating cell nuclear antigen-positive cells and the deposition of alpha-smooth muscle actin and fibronectin in the glomeruli of nephritic rats, but enalapril (30 mg/kg per day) suppressed only the number of total cells and the deposition of alpha-smooth muscle actin in the glomeruli. Moreover, to elucidate the effect of KD3-671 on matrix deposition in the glomeruli, we measured the production of fibronectin in isolated glomeruli obtained from anti-Thy-1 nephritic rats. The glomeruli in anti-Thy-1 nephritic rats produced more fibronectin than that in control rats. KD3-671 (10(-8), 10(-7), 10(-6) M) dose-dependently attenuated fibronectin production in isolated nephritic glomeruli. These findings suggest that KD3-671 may be an effective agent for the treatment of mesangioproliferative glomerulonephritis.
Assuntos
Bloqueadores do Receptor Tipo 1 de Angiotensina II , Glomerulonefrite Membranoproliferativa/tratamento farmacológico , Imidazóis/uso terapêutico , Isoanticorpos/imunologia , Glomérulos Renais , Tetrazóis/uso terapêutico , Actinas/metabolismo , Administração Oral , Animais , Anticorpos Monoclonais/imunologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Enalapril/administração & dosagem , Enalapril/farmacologia , Enalapril/uso terapêutico , Fibronectinas/metabolismo , Glomerulonefrite Membranoproliferativa/imunologia , Glomerulonefrite Membranoproliferativa/metabolismo , Glomerulonefrite Membranoproliferativa/patologia , Imidazóis/administração & dosagem , Imidazóis/farmacologia , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , Masculino , Ratos , Ratos Sprague-Dawley , Tetrazóis/administração & dosagem , Tetrazóis/farmacologiaRESUMO
1. To determine the role of actin assembly in the Ca(2+) signalling of mast cells activated by cross-linking of FcepsilonRI, we examined the effects of cytochalasin D, an inhibitor of actin polymerization. 2. In the RBL-2H3 cells, F-actin content was increased by sensitization with anti-dinitrophenol (DNP) IgE. In these cells, cytochalasin D induced oscillatory increases in cytosolic Ca(2+) ([Ca(2+)](i)); these increase were inhibited by jasplakinolide, a stabilizer of actin filaments. 3. In the IgE-sensitized RBL-2H3 cells, DNP-human serum albumin (DNP-HSA) augmented actin assembly. DNP-HSA also increased the production of IP(3), [Ca(2+)](i) and degranulation. Cytochalasin D enhanced all of these DNP-HSA-induced effects. 4. In a Ca(2+)-free solution, DNP-HSA induced a transient increase in [Ca(2+)](i), and this increase was accelerated by cytochalasin D. After cessation of the DNP-HSA-induced Ca(2+) release, the re-addition of Ca(2+) induced a sustained increase in [Ca(2+)](i) through capacitative Ca(2+) entry (CCE), and this increase was enhanced by cytochalasin D. 5 The effect of cytochalasin D in enhancing the CCE activity was prevented by xestospongin C. 6. In contrast, neither the Ca(2+) release nor the CCE activation that was induced by thapsigargin was affected by cytochalasin D. 7. These results suggest that actin de-polymerization stimulates the FcepsilonRI-mediated signalling to augment the release of Ca(2+) from the endoplasmic reticulum in RBL-2H3 cells.
Assuntos
Actinas/fisiologia , Sinalização do Cálcio , Depsipeptídeos , Mastócitos/metabolismo , Receptores de IgE/fisiologia , Actinas/metabolismo , Análise de Variância , Animais , Cátions Bivalentes , Degranulação Celular , Reagentes de Ligações Cruzadas/farmacologia , Citocalasina D/farmacologia , Dinitrofenóis/imunologia , Dinitrofenóis/farmacologia , Humanos , Imunoglobulina E/farmacologia , Inositol 1,4,5-Trifosfato/biossíntese , Compostos Macrocíclicos , Mastócitos/citologia , Oxazóis/farmacologia , Peptídeos Cíclicos/farmacologia , Ratos , Receptores de IgE/química , Albumina Sérica/imunologia , Albumina Sérica/farmacologia , Tapsigargina/farmacologia , Células Tumorais CultivadasRESUMO
1. We evaluated the role of the cross-linking of Fc epsilon RI-mediated inositol 1,4,5-triphosphate (IP(3)) in the increase in cytosolic Ca(2+) level ([Ca(2+)](i)) using xestospongin C, a selective membrane permeable blocker of IP(3) receptor, in RBL-2H3 mast cells. 2. In the cells sensitized with anti-dinitrophenol (DNP) IgE, DNP-human serum albumin (DNP-HSA) and thapsigargin induced degranulation of beta-hexosaminidase and a sustained increase in [Ca(2+)](i). Xestospongin C (3 - 10 microM) inhibited both of these changes that were induced by DNP-HSA without changing those induced by thapsigargin. 3. In the absence of external Ca(2+), DNP-HSA induced a transient increase in [Ca(2+)](i). Xestospongin C (3 - 10 microM) inhibited this increase in [Ca(2+)](i). 4. In the cells permeabilized with beta-escin, the application of IP(3) decreased Ca(2+) in the endoplasmic reticulum (ER) as evaluated by mag-fura-2. Xestospongin C (3 - 10 microM) inhibited the effect of IP(3). 5. After the depletion of Ca(2+) stores due to stimulation with DNP-HSA or thapsigargin, the addition of Ca(2+) induced capacitative calcium entry (CCE). Xestospongin C (3 - 10 microM) inhibited the DNP-HSA-induced CCE, whereas it did not affect the thapsigargin-induced CCE. 6. These results suggest that Fc epsilon RI-mediated generation of IP(3) contributes to Ca(2+) release not only in the initial phase but also in the sustained phase of the increase in [Ca(2+)](i), resulting in prolonged Ca(2+) depletion in the ER. The ER Ca(2+) depletion may subsequently activate CCE to achieve a continuous [Ca(2+)](i) increase, which is necessary for degranulation in the RBL-2H3 mast cells. Xestospongin C may inhibit Ca(2+) release and consequently may attenuate degranulation.