RESUMO
Tomatidine is isolated from the fruits of tomato plants and found to have anti-inflammatory effects in macrophages. In the present study, we investigated whether tomatidine suppresses airway hyperresponsiveness (AHR) and eosinophil infiltration in asthmatic mice. BALB/c mice were sensitized with ovalbumin and treated with tomatidine by intraperitoneal injection. Airway resistance was measured by intubation analysis as an indication of airway responsiveness, and histological studies were performed to evaluate eosinophil infiltration in lung tissue. Tomatidine reduced AHR and decreased eosinophil infiltration in the lungs of asthmatic mice. Tomatidine suppressed Th2 cytokine production in bronchoalveolar lavage fluid. Tomatidine also blocked the expression of inflammatory and Th2 cytokine genes in lung tissue. In vitro, tomatidine inhibited proinflammatory cytokines and CCL11 production in inflammatory BEAS-2B bronchial epithelial cells. These results indicate that tomatidine contributes to the amelioration of AHR and eosinophil infiltration by blocking the inflammatory response and Th2 cell activity in asthmatic mice.
Assuntos
Asma/tratamento farmacológico , Hiper-Reatividade Brônquica/tratamento farmacológico , Citocinas/imunologia , Células Th2/efeitos dos fármacos , Tomatina/análogos & derivados , Animais , Asma/imunologia , Hiper-Reatividade Brônquica/imunologia , Citocinas/análise , Modelos Animais de Doenças , Eosinófilos/efeitos dos fármacos , Eosinófilos/fisiologia , Feminino , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Células Th2/imunologia , Tomatina/farmacologia , Tomatina/uso terapêuticoRESUMO
GPR56 is a multi-functional adhesion-class G protein-coupled receptor involved in biological systems as diverse as brain development, male gonad development, myoblast fusion, hematopoietic stem cell maintenance, tumor growth and metastasis, and immune-regulation. Ectodomain shedding of human GPR56 receptor has been demonstrated previously, however the quantitative detection of GPR56 receptor shedding has not been investigated fully due to the lack of appropriate assays. Herein, an efficient system of expression and immune-affinity purification of the recombinant soluble extracellular domain of human GPR56 (sGPR56) protein from a stably transduced human melanoma cell line was established. The identity and functionality of the recombinant human sGPR56 protein were verified by Western blotting and mass spectrometry, and ligand-binding assays, respectively. Combined with the use of two recently generated anti-GPR56 monoclonal antibodies, a sensitive sandwich ELISA assay was successfully developed for the quantitative detection of human sGPR56 molecule. We found that GPR56 receptor shedding occurred constitutively and was further increased in activated human melanoma cells expressing endogenous GPR56. In conclusion, we report herein an efficient system for the production and purification of human sGPR56 protein for the establishment of a quantitative ELISA analysis of GPR56 receptor shedding.
Assuntos
Cromatografia de Afinidade/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Receptores Acoplados a Proteínas G/isolamento & purificação , Proteínas Recombinantes/isolamento & purificação , Sequência de Aminoácidos , Animais , Linhagem Celular , Eletroforese em Gel de Poliacrilamida , Vetores Genéticos/metabolismo , Humanos , Ligantes , Espectrometria de Massas , Camundongos , Dados de Sequência Molecular , Receptores Acoplados a Proteínas G/química , Proteínas Recombinantes/química , Retroviridae/metabolismo , SolubilidadeRESUMO
Proliferation of hepatic stellate cells (HSCs) plays a key role in the pathogenesis of liver fibrosis. Induction of HSC apoptosis by natural products is considered an effective strategy for treating liver fibrosis. Herein, the apoptotic effects of 7,20-epoxy-ent-kaurane (oridonin), a diterpenoid isolated from Rabdosia rubescens, and its underlying mechanisms were investigated in rat HSC cell line, HSC-T6. We found that oridonin inhibited cell viability of HSC-T6 in a concentration-dependent manner. Oridonin induced a reduction in mitochondrial membrane potential and increases in caspase 3 activation, subG1 phase, and DNA fragmentation. These apoptotic effects of oridonin were completely reversed by thiol antioxidants, N-acetylcysteine (NAC) and glutathione monoethyl ester. Moreover, oridonin increased production of reactive oxygen species (ROS), which was also inhibited by NAC. Significantly, oridonin reduced intracellular glutathione (GSH) level in a concentration- and time-dependent fashion. Additionally, oridonin induced phosphorylations of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK). NAC prevented the activation of MAPKs in oridonin-induced cells. However, selective inhibitors of MAPKs failed to alter oridonin-induced cell death. In summary, these results demonstrate that induction of apoptosis in HSC-T6 by oridonin is associated with a decrease in cellular GSH level and increase in ROS production.
Assuntos
Apoptose/efeitos dos fármacos , Diterpenos do Tipo Caurano/farmacologia , Glutationa/metabolismo , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Animais , Caspase 3/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Concentração Inibidora 50 , Espaço Intracelular/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Estresse Oxidativo , Fosforilação , Ratos , Espécies Reativas de Oxigênio/metabolismoRESUMO
OBJECTIVE: Asthma is a chronic respiratory disorder characterized by airway hyperreactivity, eosinophilic infiltration, high titer of allergen-specific IgE, and overproduction of T helper 2 (Th2) cytokines. Antigen combined with an appropriate adjuvant and administrated through the proper route can elicit suitable immunological responses to protect humans and animals from diseases. Antigen formulated with monophosphoryl lipid A (MPLA) can produce priming of Th1-mediated immune responses. The purpose of this study was to examine the utility of MPLA as an adjuvant to prevent asthma. METHODS: BALB/c mice were immunized with ovalbumin (OVA) formulated with or without MPLA by intraperitoneal, footpad, or subcutaneous injection. Vaccinated mice were challenged with OVA aerosol to estimate the protective efficacy of MPLA in comparison to Th2-adjuvant aluminum hydroxide (Alum). Airway hyperresponsiveness to methacholine, eosinophilia in bronchoalveolar lavage fluid (BALF), circulating titers of OVA-specific antibodies, and stimulating levels of IFN-γ and IL-4 cytokines from splenocytes were evaluated. RESULTS: Mice immunized by all injection routes with OVA formulated with MPLA increased the ratio of Th1/Th2 responses compared to mice receiving antigen alone. For prophylactic vaccination purpose, MPLA reduced airway responsiveness and eosinophilic inflammation in the lung, decreased serum OVA-specific IgE level, and increased the serum ratio of OVA-specific IgG2a/IgG1 and the ratio of IFN-γ/IL4 from OVA-activated splenocytes compared with mice vaccinated with Alum. CONCLUSION: MPLA may be clinically useful in the vaccination of individuals predisposed to asthma.
Assuntos
Adjuvantes Imunológicos/farmacologia , Asma/imunologia , Asma/prevenção & controle , Lipídeo A/análogos & derivados , Ovalbumina/imunologia , Células Th2/efeitos dos fármacos , Células Th2/imunologia , Animais , Testes de Provocação Brônquica/métodos , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Feminino , Imunoglobulina E/sangue , Interferon gama/imunologia , Interleucina-4/imunologia , Lipídeo A/farmacologia , Cloreto de Metacolina/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/administração & dosagem , Estatísticas não ParamétricasRESUMO
In the long history of traditional Chinese medicine, single herbs and complex formulas have been suggested to increase lifespan. However, the identification of single molecules responsible for lifespan extension has been challenging. Here, we collected a list of traditional Chinese medicines with potential longevity properties from pharmacopeias. By utilizing the mother enrichment program, we systematically screened these traditional Chinese medicines and identified a single herb, Psoralea corylifolia, that increases lifespan in Saccharomyces cerevisiae. Next, twenty-two pure compounds were isolated from Psoralea corylifolia. One of the compounds, corylin, was found to extend the replicative lifespan in yeast by targeting the Gtr1 protein. In human umbilical vein endothelial cells, RNA sequencing data showed that corylin ameliorates cellular senescence. We also examined an in vivo mammalian model, and found that corylin extends lifespan in mice fed a high-fat diet. Taken together, these findings suggest that corylin may promote longevity.
Assuntos
Células Endoteliais , Longevidade , Animais , Flavonoides/farmacologia , Mamíferos , Medicina Tradicional Chinesa , CamundongosRESUMO
Asthma is a T helper 2 (Th2) cell-associated chronic inflammatory diseases characterized with airway obstruction, increased mucus production, and eosinophil infiltration. Conventional medications for asthma treatment cannot fully control the symptoms, and potential side effects are also the concerns. Thus, complement or alternative medicine (CAM) became a new option for asthma management. Ding Chuan Tang (DCT) is a traditional Chinese herbal decoction applied mainly for patients with coughing, wheezing, chest tightness, and asthma. Previously, DCT has been proved to improve children airway hyperresponsiveness (AHR) in a randomized and double-blind clinical trial. However, the mechanisms of how DCT alleviates AHR remain unclear. Since asthmatic features such as eosinophil infiltration, IgE production, and mucus accumulation are relative with Th2 responses, we hypothesized that DCT may attenuate asthma symptoms through regulating Th2 cells. Ovalbumin (OVA) was used as a stimulant to sensitize BALB/c mice to establish an asthmatic model. AHR was detected one day before sacrifice. BALF and serum were collected for immune cell counting and antibody analysis. Splenocytes were cultured with OVA in order to determine Th2 cytokine production. Lung tissues were collected for histological and gene expression analyses. Our data reveal that DCT can attenuate AHR and eosinophil accumulation in the 30-day sensitization asthmatic model. Histological results demonstrated that DCT can reduce cell infiltration and mucus production in peribronchial and perivascular site. In OVA-stimulated splenocyte cultures, a significant reduction of IL-5 and IL-13 in DCT-treated mice suggests that DCT may alleviate Th2 responses. In conclusion, the current study demonstrates that DCT has the potential to suppress allergic responses through the reduction of mucus production, eosinophil infiltration, and Th2 activity in asthma.
Assuntos
Asma/tratamento farmacológico , Asma/imunologia , Eosinófilos/fisiologia , Imunização , Ovalbumina/imunologia , Extratos Vegetais/uso terapêutico , Pneumonia/tratamento farmacológico , Pneumonia/imunologia , Animais , Asma/sangue , Asma/fisiopatologia , Hiper-Reatividade Brônquica/sangue , Hiper-Reatividade Brônquica/complicações , Hiper-Reatividade Brônquica/fisiopatologia , Líquido da Lavagem Broncoalveolar , Regulação para Baixo , Eosinófilos/efeitos dos fármacos , Feminino , Imunoglobulina E/sangue , Interleucina-13/biossíntese , Interleucina-5/biossíntese , Camundongos Endogâmicos BALB C , Muco/metabolismo , Extratos Vegetais/farmacologia , Pneumonia/complicações , Pneumonia/fisiopatologia , Baço/patologiaRESUMO
Yin-Chen-Hao-Tang (YCHT) is recognized as a hepatoprotective agent for various types of liver diseases. Proteomics approaches were used to study hepatic and serum protein expression changes in bile duct ligated (BDL) rats following YCHT treatment for 27 days. Two-dimensional gel electrophoresis was used to analyze proteome changes. Of the proteins that exhibited changes, 17 were identified by means of matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry. The major effect of YCHT was evident in cytoskeleton related protein, plectin-1. In addition, proteins involved in metabolism of lipids were also shown to be affected, including low-density lipoprotein receptor-related protein 2 precursor (glycoprotein 330) and apolipoprotein A-I precursor (ApoA-I). Significant up-regulation of keratin 8 and 19 was found in liver tissue of BDL rats. Supplementation with YCHT also triggered alterations in the above proteins. Interestingly, YCHT treatment caused a statistically significant down-regulation in the secretion of monocyte chemoattractant protein-1 (MCP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in BDL rats with fibrosis. Our results suggested that YCHT may be useful for treatment of liver fibrosis because of its possible antiapoptotic properties, and the therapeutic effects of YCHT on liver diseases might be associated with its lipid biosynthesis regulation.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Cirrose Hepática/prevenção & controle , Fígado/efeitos dos fármacos , Proteoma/análise , Animais , Artemisia/química , Ductos Biliares/cirurgia , Proteínas Sanguíneas/análise , Combinação de Medicamentos , Medicamentos de Ervas Chinesas/química , Eletroforese em Gel Bidimensional , Gardenia/química , Expressão Gênica/efeitos dos fármacos , Ligadura/efeitos adversos , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/sangue , Cirrose Hepática/etiologia , Metaloproteinase 2 da Matriz/genética , Proteômica/métodos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rheum/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-2/genéticaRESUMO
Agaricus blazei Murill (ABM) has shown particularly strong results in treating and preventing cancer and has also traditionally been used as a food source in Brazil. However, the exact immune responses regarding the phagocytosis of macrophage and, the activity of natural killer (NK) cells in normal mice after exposure to ABM extract was unclear. The goal of this study was to investigate whether or not ABM extract can promote immune responses in normal BALB/c mice. BALB/c mice were treated with different doses of ABM extract for different time periods. The results indicated that ABM extract significantly promoted the proliferation of splenocytes both in vitro and in vivo. ABM extract promoted the levels of interleukein-6 (IL-6) and, interferon-gamma (IFN-gamma) but reduced the levels of IL-4 in vitro and in vivo. The percentage of macrophages with phagocytosis after ABM extract treatment increased and these effects were of dose-dependent manners, both in vitro and in vivo. YAC-1 target cells were killed by NK cells from the mice after treatment with ABM extract at 3 and 6 mg/kg/day for up to 14 days at target cell ratios of 25:1 and 50:1. Taken together, these results show that ABM extract promoted immunomodulations in normal BALB/c mice in vitro and in vivo.
Assuntos
Adjuvantes Imunológicos/farmacologia , Agaricus/química , Sistema Imunitário/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Linhagem Celular Transformada , Proliferação de Células/efeitos dos fármacos , Concanavalina A/farmacologia , Relação Dose-Resposta a Droga , Citometria de Fluxo , Sistema Imunitário/imunologia , Interferon gama/metabolismo , Interleucina-4/metabolismo , Interleucina-6/metabolismo , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Fagocitose/efeitos dos fármacos , Fagocitose/fisiologia , Baço/citologia , Baço/efeitos dos fármacos , Baço/imunologiaRESUMO
The increasing incidence of asthma in developing countries emphasizes the importance of identifying more effective treatments that have low cost. Gynostemma pentaphyllum (Thunb.) Makino (Cucurbitaceae), a common herbal tea in China, has been used to treat lung inflammation. Since the Th2 cytokines are the major mediators in the pathogenesis of asthma, Th1-biased immune responses caused by G. pentaphyllum might have the potential to relieve asthmatic symptoms. We hypothesized that oral administration of G. pentaphyllum extracts might suppress Th2 cytokine-induced airway inflammation responses in ovalbumin (OVA)-sensitive mice. BALB/c mice were sensitized with intraperitoneal injection and challenged 3 times with OVA inhalation (IH) (the IH3 model). G. pentaphyllum was orally administered for 7 consecutive days before the end of the OVA challenge. In the IH5 model, 2 more OVA challenges were administered to mimic the encounter with an allergen after drug treatment. G. pentaphyllum extracts significantly attenuated airway hyperresponsiveness (AHR) and inhibited eosinophil infiltration in mice in both models. Serum OVA-specific antibodies were also reduced with the treatment. Decreased Th2-type cytokines and increased IFN-gamma were detected in the cultures of OVA-activated splenocytes from treated mice. Our results suggest that G. pentaphyllum extracts might be beneficial for asthma airway inflammation through the suppression of Th2 activity.
Assuntos
Asma/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Gynostemma , Fitoterapia , Animais , Anticorpos/sangue , Asma/induzido quimicamente , Asma/metabolismo , Líquido da Lavagem Broncoalveolar/citologia , Citocinas/metabolismo , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/farmacologia , Feminino , Interleucina-5/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/efeitos adversos , Ovalbumina/imunologia , Baço/metabolismo , Células Th2/metabolismoRESUMO
BACKGROUND: Neurofibromatosis type I (NF1) is a common autosomal dominant disorder, affecting approximately one in every 3500 individuals. Early diagnosis of NF1 can be ambiguous, and clinical symptoms are diverse. We compared plasma protein profiles between normal controls and NF1 patients for yielding important insights into the mechanisms underlying NF1 related tumor formation and diagnostic biomarkers to classify the diverse clinical symptoms. METHODS: MALDI-TOF mass spectrometry was used to identify plasma proteins. Prior to that, a micro-solution isoelectric focusing (microsol-IEF) pre-fractionation combined with two-dimensional gel electrophoresis (2-DE) using the narrow pH range strip was applied to enhance the resolution and sensitivity. RESULTS: There was a significant increase in fibrinogen level in patients with NF1. This increase in fibrinogen expression was subsequently confirmed by Western blotting assay. Furthermore, the effect of fibrinogen on cell growth was tested on PC12 cells. CONCLUSION: Fibrinogen is the central protein associated with angiogenesis; a process which modulates tumor growth, the up-regulation of fibrinogen may help explain the development of neurofibromas in NF1 patients.
Assuntos
Biomarcadores/sangue , Proteínas Sanguíneas/análise , Neurofibromatose 1/sangue , Adolescente , Adulto , Estudos de Casos e Controles , Proliferação de Células , Criança , Eletroforese em Gel Bidimensional , Feminino , Humanos , Focalização Isoelétrica , Masculino , Pessoa de Meia-Idade , Neurofibromatose 1/patologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Gynostemma pentaphyllum is a popular herbal tea in China and some Asian countries. The modulatory function of G. pentaphyllum total plant extracts on immune cells was evaluated in this study. The extract was intraperitoneally injected into mice for 5 consecutive days. The production of antibodies from B cells or cytokines from T cells was determined mainly with ELISA. After the treatment, serum IgM and IgG2a were significantly enhanced and showed dose-dependent effect. Moreover, serum IgA and IgG1 were also increased when received the extract at the doses of 0.05 or 0.50 g/kg/day. In addition to the serum levels, the injection of the extract enhanced the production of all antibodies from LPS-activated spleen cells. Furthermore, more cytokines were secreted from Con A-stimulated splenocytes of G. pentaphyllum-treated mice. Our results suggest that the extract of G. pentaphyllum might promote immune responses through the activation of T and B cells.
Assuntos
Formação de Anticorpos/efeitos dos fármacos , Citocinas/biossíntese , Medicamentos de Ervas Chinesas/farmacologia , Gynostemma , Animais , Linfócitos B/imunologia , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/administração & dosagem , Injeções Intraperitoneais , Ativação Linfocitária/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Baço/citologia , Baço/imunologia , Estimulação Química , Linfócitos T/imunologiaRESUMO
Adhesion molecules expressed on cerebral endothelial cells (ECs) mediate leukocyte recruitment and play a significant role in cerebral inflammation. Increased levels of adhesion molecules on the EC surface induce leukocyte infiltration into inflammatory areas and are thus hallmarkers of inflammation. Honokiol, isolated from the Chinese medicinal herb Magnolia officinalis, has various pharmacological activities, including anti-inflammatory effects, yet the nature of honokiol targeting molecules remains to be revealed. Here, we investigated the inhibitory effect of honokiol on neutrophil adhesion and vascular cell adhesion molecule-1 (VCAM-1) expression, which underlie its molecular target, and mechanisms for inactivating nuclear factor κ enhancer binding protein (NF-κB) in mouse cerebral ECs. Honokiol inhibited tumour necrosis factor-α (TNF-α)-induced neutrophil adhesion and VCAM-1 gene expression in cerebral ECs. The inflammatory transcription factor NF-κB was downregulated by honokiol. Honokiol significantly blocked TNF-α-induced NF-κB p65 nuclear translocation and degradation of the proteasome-dependent inhibitor of NF-κB α (IκBα). From docking model prediction, honokiol directly targeted the ubiquitin-ubiquitin interface of Lys48-linked polychains. Moreover, honokiol prevented the TNF-α-induced Lys48-linked polyubiquitination, including IκBα-polyubiquitin interaction. Honokiol has protective anti-inflammatory effects on TNF-α-induced neutrophil adhesion and VCAM-1 gene expression in cerebral ECs, at least in part by directly inhibiting ubiquitination-mediated IκBα degradation and then preventing NF-κB nuclear translocation.
Assuntos
Compostos de Bifenilo/farmacologia , Encéfalo/citologia , Lignanas/farmacologia , Inibidor de NF-kappaB alfa/metabolismo , Neutrófilos/citologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Células Endoteliais/citologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Camundongos , Inibidor de NF-kappaB alfa/química , Neutrófilos/efeitos dos fármacos , Proteólise/efeitos dos fármacos , Ubiquitinação/efeitos dos fármacos , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
Thyroid hormones regulate growth, development, differentiation, and metabolic processes by interacting with and activating thyroid hormone receptors and associated pathways. We investigated the triiodothyronine (T3) modulation of gene expression, in human hepatocellular carcinoma cell lines, via a PCR-based cDNA subtraction method. Here we present further data on one of the T3-upregulated genes, fibronectin (FN). We demonstrate that the induction of FN protein expression by T3 in TRalpha1 and TRbeta1 over-expressing cells was time and dose-dependent at the mRNA and protein levels. Blockade of protein synthesis by cycloheximide almost completely inhibited the concomitant induction of FN mRNA by T3, indicating that T3 indirectly regulates FN. Furthermore, nuclear-run on and FN promoter assay clearly can specifically increase the number of FN transcriptional demonstrated that the presence of T3 initiations. In addition, we further confirmed that the up-regulation of FN by T3 was mediated, at least in part, by transforming growth factor-beta (TGF-beta), because the induction of FN was blocked in a dose-dependent manner by the addition of TGF-beta neutralizing antibody. In an effort to elucidate the we demonstrated the involvement of the signaling pathways involved in the activation of FN by T3, mitogen activated protein kinase/c-Jun N-terminal kinase/p38 MAPK (MAPK/JNK/p38) pathway. Although T3 induces the expression of TGF-beta, neither wild-type nor dominant-negative Smad3 or Smad4 over-expression affected the activation of FN by T3. Thus, we demonstrate that T3 regulates FN gene expression indirectly at the transcriptional level, with the participation of the MAPK/JNK/p38 pathway and the TGF-beta signaling pathway but independent of Smad3/4.
Assuntos
Fibronectinas/metabolismo , Receptores alfa dos Hormônios Tireóideos/metabolismo , Receptores beta dos Hormônios Tireóideos/metabolismo , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Cicloeximida/farmacologia , Proteínas de Ligação a DNA/efeitos dos fármacos , Proteínas de Ligação a DNA/metabolismo , Inibidores Enzimáticos/farmacologia , Fibronectinas/efeitos dos fármacos , Fibronectinas/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Proteína Smad3 , Proteína Smad4 , Receptores alfa dos Hormônios Tireóideos/efeitos dos fármacos , Receptores alfa dos Hormônios Tireóideos/genética , Receptores beta dos Hormônios Tireóideos/efeitos dos fármacos , Receptores beta dos Hormônios Tireóideos/genética , Transativadores/efeitos dos fármacos , Transativadores/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Tri-Iodotironina/metabolismo , Tri-Iodotironina/farmacologia , Células Tumorais CultivadasRESUMO
Liu-Wei-Di-Huang Wan (LWDHW) has been used by traditional Chinese doctors to treat asthma patients. This study was to examine the potential effect of this decoction on the regulation of T helper (Th)1- and Th2-type cytokine gene expression in vitro. Peripheral blood mononuclear cells (PBMC) were activated with mitogen for 24 hours in the presence or absence of LWDHW extracts. Concentrations of different cytokines in the culture supernatants were determined with ELISA. RNA isolated from cultured cells was subjected to RT-PCR analysis. The results showed that the expression of all cytokines (Th2-type: IL-4, IL-5, IL-10, or IL-13 and Th1-type: IL-2 and IFN-gamma) examined was inhibited at both RNA and protein levels by LWDHW. Since the cell viability was similar in all cultures, the reduction of cytokine production was not due to the toxicity of LWDHW. Moreover, the cells either retained or increased their capacity to respond to mitogen stimulation after incubation with the LWDHW decoction. Therefore, the data suggest that LWDHW functioned directly on cytokine gene expression from activated PBMC.
Assuntos
Antiasmáticos/farmacologia , Citocinas/biossíntese , Medicamentos de Ervas Chinesas/farmacologia , Linfócitos/metabolismo , Animais , Asma/patologia , Divisão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Criança , Citocinas/genética , Expressão Gênica/efeitos dos fármacos , Humanos , Linfócitos/efeitos dos fármacos , Ácaros/imunologia , Mitógenos/farmacologia , RNA/biossíntese , RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Testes Cutâneos , Células Th2/efeitos dos fármacos , Células Th2/imunologiaRESUMO
PURPOSE: Lovastatin is an effective inhibitor of cholesterol synthesis. A previous study demonstrated that lovastatin can also suppress airway hyperresponsiveness (AHR) in murine model of asthma. We aimed to investigate the effect of lovastatin on mucus secretion and inflammation-associated gene expression in the lungs of murine model of asthma. METHODS: Female BALB/c mice were sensitized and challenged with ovalbumin (OVA) by intraperitoneal injection, and orally administered lovastatin from days 14 to 27 post-injection. Gene expression in lung tissues was analyzed using real-time polymerase chain reaction. AHR and goblet cell hyperplasia were also examined. BEAS-2B human bronchial epithelial cells were used to evaluate the effect of lovastatin on the expression of cell adhesion molecules, chemokines, and proinflammatory cytokines in vitro. RESULTS: We showed that lovastatin inhibits the expression of Th2-associated genes, including eotaxins and adhesion molecules, in the lungs of murine model of asthma. Mucin 5AC expression, eosinophil infiltration and goblet cell hyperplasia were significantly decreased in the lung tissue of murine model of asthma treated with lovastatin. Furthermore, lovastatin inhibited AHR and expression of Th2-associated cytokines in bronchoalveolar lavage fluid. However, a high dose (40 mg/kg) of lovastatin was required to decrease specific IgE to OVA levels in serum, and suppress the expression of Th2-associated cytokines in splenocytes. Activated BEAS-2B cells treated with lovastatin exhibited reduced IL-6, eotaxins (CCL11 and CCL24), and intercellular adhesion molecule-1 protein expression. Consistent with this, lovastatin also suppressed the ability of HL-60 cells to adhere to inflammatory BEAS-2B cells. CONCLUSIONS: These data suggest that lovastatin suppresses mucus secretion and airway inflammation by inhibiting the production of eotaxins and Th2 cytokines in murine model of asthma.
RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Matrine has been isolated from Sophora flavescens, and found to show anti-inflammatory effects in macrophages and anti-cachectic effects in hepatomas. The present study investigated whether matrine suppressed eosinophil infiltration and airway hyperresponsiveness (AHR) in mice, and decreased the inflammatory response of tracheal epithelial cells. MATERIALS AND METHODS: BALB/c mice were sensitized and challenged with ovalbumin to induce allergic asthma in mice. These asthmatic mice were given various doses of matrine by intraperitoneal injection. Additionally, activated human tracheal epithelial cells (BEAS-2B cells) were treated with matrine, and evaluated for levels of proinflammatory cytokines and chemokines. RESULTS: We found that matrine significantly decreased AHR, and suppressed goblet cell hyperplasia, eosinophil infiltration, and inflammatory response in the lung tissue of asthmatic mice. Matrine also reduced the levels of Th2 cytokines and chemokines in bronchoalveolar lavage fluid, and suppressed OVA-IgE production in serum. Furthermore, matrine treatment of activated BEAS-2B cells decreased production of proinflammatory cytokines and eotaxins, as well as suppressed ICAM-1 expression and thus adhesion of eosinophils to inflammatory BEAS-2B cells in vitro. CONCLUSIONS: Our findings suggest that matrine can improve allergic asthma in mice, and therefore has potential therapeutic potential in humans.
Assuntos
Alcaloides/farmacologia , Asma/tratamento farmacológico , Citocinas/metabolismo , Eosinófilos/efeitos dos fármacos , Quinolizinas/farmacologia , Animais , Asma/imunologia , Adesão Celular , Linhagem Celular , Citocinas/genética , Eosinófilos/fisiologia , Regulação da Expressão Gênica , Células Caliciformes/efeitos dos fármacos , Humanos , Hipersensibilidade , Pulmão/citologia , Pulmão/efeitos dos fármacos , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina , MatrinasRESUMO
BACKGROUND: Th2 cells are overexpressed in the skin and serum of atopic dermatitis (AD) patients. Previously, we found that dehydroepiandrosterone (DHEA) decreased eosinophil infiltration in asthmatic mice through the suppression of Th2-associated cytokines. Therefore, we hypothesized that DHEA might improve the symptoms of AD syndrome. OBJECTIVE: In this study, we evaluated the symptom improvement and anti-inflammatory response that result from the modulation of immunity by DHEA modulated in AD-like mice. METHODS: Female BALB/c mice were sensitized and challenged with 1-chloro-2,4-dinitrobenzene. On days 14-29 after sensitization, mice were treated with cutaneous (skin smear) or oral administration of DHEA. In addition, human keratinocyte (HaCat) cells were used to evaluate the effect of DHEA on the in vitro production of proinflammatory cytokines and chemokines. RESULTS: Both cutaneous and oral DHEA were able to decrease ear swelling and skin inflammation in AD-like mice. DHEA also attenuated eosinophil and mast cell infiltration into ear and skin tissue. Additionally, Th2-associated cytokines were inhibited in splenocyte culture, and suppressed the levels of IgE and interleukin 4 in serum. Oral and cutaneous administration of DHEA reduced the inflammatory response, as evidenced by AD-like skin lesions, in a similar manner. DHEA significantly reduced inflammatory cytokines and chemokines through the nuclear factor-κB and mitogen-activated protein kinases pathways in tumor necrosis factor-α activated HaCat cells. CONCLUSION: DHEA ameliorates AD-like mouse skin inflammation and reduces eosinophil and mast cell infiltration by reducing the production of Th2-associated cytokines and chemokines.
Assuntos
Adjuvantes Imunológicos/farmacologia , Desidroepiandrosterona/farmacologia , Dermatite Atópica/tratamento farmacológico , Dinitroclorobenzeno/farmacologia , Pele/efeitos dos fármacos , Animais , Anti-Inflamatórios/farmacologia , Quimiocinas/metabolismo , Citocinas/metabolismo , Eosinófilos/metabolismo , Feminino , Humanos , Inflamação , Queratinócitos/efeitos dos fármacos , Mastócitos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Fosforilação , Baço/citologia , Células Th2/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
AIM OF THE STUDY: Viscum coloratum Nakai is used in traditional Chinese medicine to treat various diseases, including hemorrhage, hypertension, and inflammatory diseases. A previous study demonstrated a partially purified extract (PPE-SVC) and viscolin from Viscum coloratum Nakai inhibited phosphodiesterase activity. In this study, we evaluated the anti-asthmatic effects of PPE-SVC and viscolin, from Viscum coloratum Nakai, in OVA-sensitized mice. MATERIALS AND METHODS: Female BALB/c mice were sensitized and challenged with ovalbumin (OVA). The mice were randomized into groups and treated with PPE-SVC, viscolin, or rolipram by intraperitoneal injection on 1h before each inhalation of OVA and airway hyperresponsiveness (AHR). RESULTS: PPE-SVC and viscolin suppressed AHR and reduced eosinophil infiltration of the lungs in OVA-sensitized mice. Moreover, PPE-SVC and viscolin inhibited chemokines, including CCL11 and CCL24, and Th2-associated cytokines in bronchoalveolar lavage fluid. However, PPE-SVC and viscolin could not decrease IL-4, IL-5, and IL-13 levels in cultures of OVA-activated spleen cells. CONCLUSION: PPE-SVC and viscolin attenuate airway inflammation and eosinophil infiltration in OVA-sensitized mice.
Assuntos
Antiasmáticos/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Compostos de Bifenilo/uso terapêutico , Hiper-Reatividade Brônquica/prevenção & controle , Eosinófilos/metabolismo , Extratos Vegetais/uso terapêutico , Propano/análogos & derivados , Viscum/química , Animais , Antiasmáticos/isolamento & purificação , Antiasmáticos/farmacologia , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Compostos de Bifenilo/isolamento & purificação , Compostos de Bifenilo/farmacologia , Hiper-Reatividade Brônquica/imunologia , Hiper-Reatividade Brônquica/metabolismo , Líquido da Lavagem Broncoalveolar/química , Quimiocinas/metabolismo , Citocinas/metabolismo , Feminino , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/prevenção & controle , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Medicina Tradicional Chinesa , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina , Fitoterapia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Propano/isolamento & purificação , Propano/farmacologia , Propano/uso terapêutico , Distribuição Aleatória , Rolipram , Baço/citologia , Baço/efeitos dos fármacosRESUMO
In a variety of countries, juvenile idiopathic arthritis (JIA) is the most common cause of chronic arthritis in childhood, yet its etiology is still unknown. In recent years, etanercept, an effective inhibitor of tumor necrosis factor alpha (TNF-alpha), was used as an alternative in certain oligoarticular JIA patients resistant to conventional nonsteroidal anti-inflammatory drugs (NSAIDs), disease-modifying anti-rheumatic drugs (DMARDs), and corticosteroid therapies, and it resulted in sustained improvement in JIA symptoms. This pilot study explores the alterations of specific panels of cytokines and protein profiles in plasma for two Taiwanese pediatric cases with diagnosed enthesitis-related arthritis (ERA), a type of JIA. The patients were studied before and after taking etanercept alone, using a high-content screening approach employing membrane-based human cytokine antibody microarray and the conventional two-dimensional gel electrophoresis (2-DE) proteomic technique. Specifically, 2-DE in combination with mass spectrometry (MALDI-MS) revealed the functional roles of plasma proteins associated with the regulation of immune responses during short-term etanercept treatment of children with ERA. Our study shows that this biotherapy improved clinical ERA manifestations through the regulation of inflammatory mediators, including several cytotoxic cellular cytokines (IL-2/IFN-g), chemokines (MCP-1), and growth factors (GRO) that affect the expression of specific acute phase proteins such as haptoglobins, immunoglobulin A, and fibrinogen-gamma chain. Meanwhile, an up-regulation of antithrombin chain I, vitamin-D binding protein (VDBP), and the various apolipoproteins was also observed after the administration of etanercept in both studied children. These results may be interpreted as the relevant predictive biomarkers of therapeutic responses to etanercept. They suggest that etanercept, which is still rarely used in Taiwan, is a viable treatment for JIA patients, without adverse health effects and increased risk of secondary infections.
Assuntos
Artrite Juvenil/sangue , Artrite Juvenil/tratamento farmacológico , Proteínas Sanguíneas/análise , Imunoglobulina G/uso terapêutico , Proteômica , Receptores do Fator de Necrose Tumoral/uso terapêutico , Artrite Juvenil/imunologia , Biomarcadores/sangue , Proteínas Sanguíneas/imunologia , Criança , Eletroforese em Gel Bidimensional , Etanercepte , Ensaios de Triagem em Larga Escala , Humanos , Imunoglobulina G/administração & dosagem , Masculino , Projetos Piloto , Valor Preditivo dos Testes , Receptores do Fator de Necrose Tumoral/administração & dosagem , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Our previous report demonstrated that the oral administration of short-term high dose Gynostemma pentaphyllum extract (5 g/kg per day for 7days) decreased allergic reactions in ovalbumin (OVA)-sensitized mice. The aim of this study was to determine whether long-term oral administration of G. pentaphyllum attenuated airway inflammation in OVA-sensitized mice. Mice were sensitized and challenged with normal saline or OVA. OVA-sensitized mice were fed with 1.75 g/kg (low dose, GPL) or 5 g/kg (high dose, GPH) G. pentaphyllum extract, five days a week for 4 weeks. The airway hyperresponsiveness (AHR) and eosinophilia in bronchoalveolar lavage fluid (BALF) were examined. The cytokine levels or antibodies in BALF, serum and spleen cell culture supernatants were also determined. Both high and low dose extracts reduced AHR, serum OVA-IgE, and Th2-associated cytokine levels in spleen cell supernatants and BALF in OVA-sensitized mice. These results show that long-term orally administered G. pentaphyllum extract reduced allergic reactions in OVA-sensitized mice.