RESUMO
We found that a water-soluble extract of coriander (Coriandrum sativum L.) (leaves, petioles and stems) inhibits antigen-induced degranulation of RBL-2H3 cells, a rat basophil leukemia cell line. The aim of this study was to elucidate the anti-degranulation active components in the extract. The methanol-eluate fraction obtained by fractionation of the water-soluble extract using MCI gel column chromatography had strong activity, and eight components were isolated and identified. Two of them were identified as new compounds, (3S)-3-methyl-6-hydroxyisocoumarin 8-O-ß-D-glucopyranoside (compound 1) and (7S,8R)-7,8-dihydro-8-ß-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-fro[2,3-g][2]benzopyran-5-one (compound 2). As a result of evaluation of anti-degranulation activity of eight components, seven of them, such as tryptophan, phenylalanine, dihydroxycoumarin glucoside, quercetin glycoside, rutin, compound 1, and compound 2, had the activity. These results indicated that the water-soluble extract of coriander contains several anti-degranulation substances.
Assuntos
Coriandrum , Animais , Ratos , Estrutura Molecular , Extratos Vegetais/química , Folhas de Planta/química , Rutina , ÁguaRESUMO
Cuminum cyminum L. (cumin) is an annual plant of the Umbelliferae family native to Egypt. We previously showed that the aqueous extract of cumin seeds suppresses degranulation by downregulating the activation of antigen-induced intracellular signaling molecules in rat basophilic leukemia RBL-2H3 cells. However, the active substances in the extract have not yet been identified. Accordingly, herein, we aimed to ascertain the water-soluble substances present in cumin seeds that inhibit degranulation, which led to the identification of umbelliferose, a characteristic trisaccharide present in plants of the Umbelliferae family. Our study is the first to reveal the degranulation-suppressing activity of umbelliferose, and quantification studies suggest that cumin seed powder contains 1.6% umbelliferose. Raffinose, an isomer of umbelliferose, was also found to significantly suppress antigen-induced degranulation, but less so than umbelliferose. Both umbelliferose and raffinose contain sucrose subunits in their structures, with galactose moieties bound at different sites. These differences in structure suggest that the binding of galactose to the sucrose subunit at the α1-2 bond contributes to its strong degranulation-inhibiting properties.
Assuntos
Cuminum , Leucemia , Animais , Degranulação Celular , Cuminum/química , Galactose/análise , Extratos Vegetais/química , Rafinose/análise , Ratos , Sementes/química , Sacarose/análiseRESUMO
Trigonelline, one of the alkaloids contained in coffee, is important not only as one of the constituents of aroma and flavor in coffee but also as a useful source of nutrition. Its anti-microbial, anti-carcinogenic, and anti-hyperglycemic effects have been investigated in previous studies. However, there have not been any studies examining the anti-degranulation effect of trigonelline. In this study, the anti-degranulation effect of trigonelline was evaluated in in vitro and in vivo models using a rat basophilic leukemia cell line, RBL-2H3 cells, and a passive cutaneous anaphylaxis (PCA) reaction in mice, respectively. In the ß-hexosaminidase release assay, trigonelline effectively suppressed antigen-induced degranulation of RBL-2H3 cells in a dose-dependent manner without cytotoxicity. Trigonelline also inhibited FcεRI-mediated intracellular signaling pathways, such as phosphorylation of PLCγ1, PI3 K, and Akt, in antigen-stimulated RBL-2H3 cells and suppressed the PCA response in mice. Moreover, trigonelline also inhibited the microtubule formation in RBL-2H3 cells, indicating that trigonelline could inhibit IgE-sensitized mast cell degranulation by attenuating both the intracellular calcium-dependent and independent pathways. These results revealed that trigonelline possesses the anti-degranulation effect against the development of allergic diseases.
Assuntos
Alcaloides/farmacologia , Degranulação Celular/efeitos dos fármacos , Animais , Antialérgicos/farmacologia , Cálcio/metabolismo , Linhagem Celular Tumoral , Feminino , Hipersensibilidade/tratamento farmacológico , Hipersensibilidade/metabolismo , Imunoglobulina E/metabolismo , Leucemia/tratamento farmacológico , Leucemia/metabolismo , Mastócitos/efeitos dos fármacos , Mastócitos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Anafilaxia Cutânea Passiva/efeitos dos fármacos , Extratos Vegetais/farmacologia , Ratos , Transdução de Sinais/efeitos dos fármacosRESUMO
Immunoglobulin E (IgE) is involved in the onset of allergic reaction, and the suppression of IgE production leads to alleviation of allergic symptoms. We found that mango peel ethanol extract (MPE) significantly suppresses IgE production by human myeloma cell line U266 cells, suggesting that MPE has an anti-allergic effect by inhibiting the production of IgE. Although mangiferin is contained in mango, which suppresses IgE production by U266 cells, it was not contained in MPE. We investigated the suppressive effect of MPE in 2,4-dinitrofluorobenzene (DNFB)-induced allergic contact dermatitis model mice. The elevation of serum IgE level was significantly suppressed by oral administration of MPE. Intake of MPE also suppressed the expression level of IL-4 in the DNFB-challenged ears, suggesting that MPE suppresses the IL-4-mediated maturation into IgE-producing cells. Our findings indicate that MPE has a potential to alleviate the increase in serum IgE level that is feature of type I allergy.
Assuntos
Etanol/química , Imunoglobulina E/biossíntese , Mangifera/química , Extratos Vegetais/farmacologia , Animais , Linhagem Celular Tumoral , Dermatite Alérgica de Contato/imunologia , Dinitrobenzenos/toxicidade , Modelos Animais de Doenças , Orelha , Expressão Gênica/efeitos dos fármacos , Humanos , Switching de Imunoglobulina , Imunoglobulina E/sangue , Imunoglobulina E/genética , Interleucina-4/genética , Camundongos Endogâmicos BALB CRESUMO
Purpose: To evaluate the effects of mandarin orange yogurt containing nobiletin and ß-lactoglobulin on the allergic conjunctivitis induced by a conjunctival allergen challenge (CAC). Methods: Experiment 1 was performed on 26 asymptomatic patients (age, 25.3 ± 5.3 years) with proven seasonal allergic conjunctivitis due to cedar pollen. We compared the degree of conjunctivitis induced by CAC before and after ingesting mandarin orange yogurt for 2 weeks. Experiment 2 was a double-blind, placebo-controlled trial performed on 31 patients (age, 32.5 ± 12.2 years). A diet containing mandarin orange yogurt was compared to a diet containing yogurt lacking the mandarin orange on the conjunctivitis induced by CAC. The temperature of the inferior bulbar conjunctiva was measured before and 20 minutes after the CAC with an ocular surface thermographer (OST). The degree of conjunctival injection and chemosis was graded by slit-lamp biomicroscopy. The changes in the symptoms were evaluated by a questionnaire. Results: In experiment 1, the scores of redness (3.07 ± 3.03 vs. 1.05 ± 1.70), chemosis (2.84 ± 2.27 vs. 0.81 ± 1.11), itching (4.34 ± 3.05 vs. 1.39 ± 2.12), and temperature (0.73 ± 0.42°C vs. 0.45 ± 0.43°C) were significantly lower (P < 0.001) after a diet of mandarin orange yogurt for 2 weeks. In experiment 2, the scores of redness (1.03 ± 0.18 vs. 1.28 ± 0.52; P = 0.0156), itching (1.93 ± 1.92 vs. 2.82 ± 2.21; P = 0.0133), and surface temperature (0.54 ± 0.21°C vs. 0.31 ± 0.25°C; P < 0.001) were significantly lower in the mandarin orange yogurt group than in the control yogurt group. Conclusions: Mandarin orange yogurt can be an effective nutritional intervention for allergic conjunctivitis.
Assuntos
Alérgenos/imunologia , Citrus , Túnica Conjuntiva/patologia , Conjuntivite Alérgica/tratamento farmacológico , Óleos de Plantas/farmacologia , Iogurte , Adulto , Túnica Conjuntiva/efeitos dos fármacos , Conjuntivite Alérgica/imunologia , Conjuntivite Alérgica/patologia , Feminino , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade , Soluções Oftálmicas , Adulto JovemRESUMO
BACKGROUND: Coriandrum sativum L. seed is generally used as a spice and crude drug. Although many functions of the various components in C. sativum L. seed have been reported, the immunostimulatory effect of water-soluble components in C. sativum L. seed has not been studied. In the present study, we focused on the immunostimulatory effect of C. sativum L. seed aqueous extract (CAE) on macrophages as a novel health function of C. sativum L. seed components. RESULTS: CAE significantly enhanced the production of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in both RAW264.7 cells and peritoneal macrophages by enhancing the expression levels of these cytokine genes. CAE also stimulated nitric oxide (NO) production and the phagocytosis activity in RAW264.7 cells. We suggest that the activity of CAE is a result of the upregulation of mitogen-activated protein kinase and nuclear factor-κB cascades via TLR4. In addition, IL-6 production by peritoneal macrophages collected from CAE-administered mice was significantly enhanced, suggesting that CAE could stimulate macrophage activity in vivo. CONCLUSION: The findings of the present study suggest that CAE contains a novel water-soluble component with an immunostimulatory effect on macrophages. CAE would contribute to activating host defense against pathogens by stimulating the innate immunity. © 2017 Society of Chemical Industry.
Assuntos
Adjuvantes Imunológicos , Coriandrum/química , Imunidade/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Extratos Vegetais/farmacologia , Animais , Expressão Gênica/efeitos dos fármacos , Interleucina-6/biossíntese , Interleucina-6/genética , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/imunologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Óxido Nítrico/biossíntese , Fagocitose/efeitos dos fármacos , Células RAW 264.7 , Sementes/química , Solubilidade , Receptor 4 Toll-Like/fisiologia , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/genética , ÁguaRESUMO
Chlorophyll c2 extracted from Sargassum horneri improved allergic symptoms in an animal model of allergic rhinitis. In the present study, we explored the efficacy of chlorophyll c2 in patients with seasonal allergic rhinitis. This was a single-center, randomized, double-blind placebo-controlled trial. Sixty-six patients aged 20-43 years, each with a 2-year history of seasonal allergic rhinitis, were randomly assigned to receive either a single daily dose (0.7 mg) of chlorophyll c2 or placebo for 12 weeks. The use of medications including H1-antihistamines and topical nasal steroids was recorded by rescue medication scores (RMSs) noted after 4, 8, and 12 weeks of treatment. Disease-specific quality of life was measured using the Japan Rhinitis Quality of Life Questionnaire (JRQLQ) both before and after 4, 8, and 12 weeks of treatment. The RMS at 8 weeks was significantly better in the chlorophyll c2 than the placebo group (mean RMS difference = -3.09; 95 % confidence interval = -5.96 to -0.22); the mean RMS at 4 weeks was only slightly better in the chlorophyll c2 group. The JRQLQ scores did not differ significantly between the two groups. Chlorophyll c2 would have a potential to be an alternative treatment for allergic rhinitis.
Assuntos
Clorofila/uso terapêutico , Rinite Alérgica Sazonal/terapia , Sargassum , Adulto , Método Duplo-Cego , Feminino , Humanos , Japão , Masculino , Qualidade de Vida , Método Simples-Cego , Inquéritos e Questionários , Fatores de Tempo , Adulto JovemRESUMO
We herein report the immunostimulatory effect of spinach aqueous extract (SAE) on mouse macrophage-like J774.1 cells and mouse primary peritoneal macrophages. SAE significantly enhanced the production of interleukin (IL)-6 and tumor necrosis factor-α by both J774.1 cells and peritoneal macrophages by enhancing the expression levels of these cytokine genes. In addition, the phagocytosis activity of J774.1 cells was facilitated by SAE. Immunoblot analysis revealed that SAE activates mitogen-activated protein kinase and nuclear factor-κB cascades. It was found that SAE activates macrophages through not only TLR4, but also other receptors. The production of IL-6 was significantly enhanced by peritoneal macrophages from SAE-administered BALB/c mice, suggesting that SAE has a potential to stimulate macrophage activity in vivo. Taken together, these data indicate that SAE would be a beneficial functional food with immunostimulatory effects on macrophages.
Assuntos
Adjuvantes Imunológicos/farmacologia , Interleucina-6/agonistas , Macrófagos Peritoneais/efeitos dos fármacos , Extratos Vegetais/farmacologia , Spinacia oleracea/química , Fator de Necrose Tumoral alfa/agonistas , Animais , Linhagem Celular , Feminino , Expressão Gênica , Interleucina-6/genética , Interleucina-6/imunologia , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/agonistas , NF-kappa B/genética , NF-kappa B/imunologia , Fagocitose/efeitos dos fármacos , Cultura Primária de Células , RNA Mensageiro/agonistas , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
BACKGROUND: Equol is a metabolite of daidzein that is produced by intestinal microbiota. The oestrogenic activity of equol is stronger than daidzein. Equol-producing bacteria are believed to play an important role in the gut. The rod-shaped and Gram-positive anaerobic equol-producing intestinal bacterium Slackia TM-30 was isolated from healthy human faeces and its effects on urinary phyto-oestrogen, plasma and faecal lipids were assessed in adult mice. RESULTS: The urinary amounts of equol in urine were significantly higher in mice receiving the equol-producing bacterium TM-30 (BAC) group than in the control (CO) group (P < 0.05). However, no significant differences were observed between the urinary amounts of daidzein, dihydrodaidzein, enterodiol, and enterolactone between the BAC and CO groups. No significant differences in the plasma lipids were observed between the two groups. The lipid content (% dry weight) in the faeces sampled on the final day of the experiment tended to be higher in the BAC group than in the CO group (P = 0.07). CONCLUSION: Administration of equol-producing bacterium TM-30 affected the urinary amounts of phyto-oestrogens and the faecal lipid contents of mice. The equol-producing bacterium TM-30 likely influences the metabolism of phyto-oestrogen via changes in the gastrointestinal environment. © 2015 Society of Chemical Industry.
Assuntos
Actinobacteria/metabolismo , Equol/biossíntese , Fezes/microbiologia , Microbioma Gastrointestinal , Isoflavonas/metabolismo , 4-Butirolactona/análogos & derivados , 4-Butirolactona/urina , Actinobacteria/isolamento & purificação , Animais , Equol/administração & dosagem , Equol/urina , Fezes/química , Feminino , Humanos , Isoflavonas/urina , Lignanas/metabolismo , Lignanas/urina , Lipídeos/sangue , Camundongos , Camundongos Endogâmicos ICR , Fitoestrógenos/urinaRESUMO
Bengkoang (Pachyrhizus erosus (L.) Urban) is an edible root tuber containing fairly large amounts of carbohydrates and crude fibers. Our previous studies showed that the bengkoang fiber extract (BFE) stimulates activation of macrophages, leading to induction of phagocytotic activity and cytokine production. In the present study we investigated the mechanism underlying activation of murine macrophages by BFE. BFE increased production of TNF-α, IL-6, and nitric oxide by J774.1 cells. In addition BFE also facilitated the gene expression levels of inducible nitric oxide synthase. We examined the effect of a TLR4 inhibitor on cytokine production to investigate the membrane receptor of macrophage activation by BFE. Treatment of J774.1 cells with the TLR4 inhibitor significantly inhibited production of IL-6 and TNF-α, suggesting that TLR4 is the target membrane receptor for BFE. The main signal molecules located downstream of TLR4 such as JNK, p38, ERK, and NF-κB were activated by BFE treatment. The immunostimulatory effect of BFE was cancelled by the pectinase treatment, suggesting that the active ingredient in BFE is pectin-like molecules. Overall results suggested that BFE activates J774.1 cells via the MAPK and NF-κB signaling pathways.
Assuntos
Fibras na Dieta/farmacologia , Macrófagos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Receptor 4 Toll-Like/metabolismo , Animais , Linhagem Celular , Imunomodulação , Interleucina-6/metabolismo , Lipopolissacarídeos/efeitos adversos , Ativação de Macrófagos/efeitos dos fármacos , Camundongos , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Poligalacturonase/metabolismo , Transdução de Sinais , Receptor 4 Toll-Like/antagonistas & inibidores , Receptor 4 Toll-Like/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
We previously reported that jellyfish collagen stimulates both the acquired and innate immune responses. In the acquired immune response, jellyfish collagen enhanced immunoglobulin production by lymphocytes in vitro and in vivo. Meanwhile, in the innate immune response jellyfish collagen promoted cytokine production and phagocytotic activity of macrophages. The facts that jellyfish collagen plays several potential roles in stimulating cytokine production by macrophages have further attracted us to uncover its mechanisms. We herein describe that the cytokine production-stimulating activity of jellyfish collagen was canceled by a Toll-like receptor 4 (TLR4) inhibitor. Moreover, jellyfish collagen stimulated phosphorylation of inhibitor of κBα (IκBα), promoted the translocation of nucleus factor-κB (NF-κB), and activated c-Jun N-terminal kinase (JNK). A JNK inhibitor also abrogated the cytokine production-stimulating activity of jellyfish collagen. These results suggest that jellyfish collagen may facilitate cytokine production by macrophages through activation of NF-κB and JNK via the TLR4 signaling pathways.
Assuntos
Colágeno/imunologia , Proteínas Quinases JNK Ativadas por Mitógeno/imunologia , Macrófagos/imunologia , NF-kappa B/imunologia , Receptor 4 Toll-Like/imunologia , Animais , Antracenos/farmacologia , Linhagem Celular , Colágeno/farmacologia , Endotoxinas/imunologia , Proteínas I-kappa B/metabolismo , Interleucina-6/biossíntese , Proteínas Quinases JNK Ativadas por Mitógeno/antagonistas & inibidores , Camundongos , Inibidor de NF-kappaB alfa , Fosforilação , Cifozoários/imunologia , Cifozoários/metabolismo , Receptor 4 Toll-Like/antagonistas & inibidores , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/biossínteseRESUMO
The cytotoxic activities of sesquilignans, (7S,8S,7'R,8'R)- and (7R,8R,7'S,8'S)-morinol A and (7S,8S,7'S,8'S)- and (7R,8R,7'R,8'R)-morinol B were compared, showing no significant difference between stereoisomers (IC50=24-35 µM). As a next stage, the effect of substituents at 7, 7', and 7"-aromatic ring on the activity was evaluated to find out the higher activity of (7S,8S,7'R,8'R)-7,7',7"-phenyl derivative 18 (IC50=6-7 µM). In the research on the structure-activity relationship of 7"-position of (7S,8S,7'R,8'R)-7,7',7"-phenyl derivative 18, the most potent compounds were 7,7',7"-phenyl derivative 18 (IC50=6 µM) against HeLa cells. Against HL-60 cells, 7"-(4-nitrophenyl)-7,7'-phenyl derivative 33 and 7"-hexyl-7,7'-phenyl derivative 37 (IC50=5 µM) showed highest activity. We discovered the compounds showed four to sevenfold potent activity than that of natural (7S,8S,7'R,8'R)-morinol A. It was also confirmed that the 7'-benzylic hydroxy group have an important role for exhibiting activity, on the other hand, the resonance system of cinnamyl structure is not crucial for the potent activity.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Lignanas/farmacologia , Piranos/farmacologia , Antineoplásicos Fitogênicos/química , Sobrevivência Celular/efeitos dos fármacos , Medicamentos de Ervas Chinesas/química , Células HL-60 , Células HeLa , Humanos , Lignanas/química , Neoplasias/tratamento farmacológico , Piranos/química , Relação Estrutura-AtividadeRESUMO
The inhibitory effect of an aqueous extract from spinach on degranulation of RBL-2H3 cells is herein reported. The extract significantly suppressed antigen-induced degranulation in a dose-dependent manner without affecting cell viability. Active substances in the extract were heat-stable and trypsin-resistant with molecular weights ranging from 500 Da to 14 kDa. The extract inhibited elevation of the intracellular Ca(2+) concentration caused by stimulation by antigen, while not suppressing degranulation induced by a calcium ionophore A23187. Immunoblot analysis revealed that the inhibitory effect results from downregulation of phosphorylation of both Syk kinase and phosphatidylinositol 3-kinase in the signalling pathways involved in degranulation caused by the antigen-antibody interaction. Taken together, these findings suggest that aqueous spinach extract has an anti-allergic activity that controls degranulation.
Assuntos
Antialérgicos/farmacologia , Basófilos/efeitos dos fármacos , Degranulação Celular/efeitos dos fármacos , Extratos Vegetais/farmacologia , Spinacia oleracea/química , Animais , Basófilos/imunologia , Cálcio/imunologia , Linhagem Celular Tumoral , RatosRESUMO
Heavy oil is one of the most serious pollutants in marine ecosystem. The poisonous influences of the chemical substances contained in heavy oil on many kinds of marine organisms are widely studied. However, the influence of the chemical compounds in heavy oil on our health has not been cleared yet. In order to reveal the poisonous influences of these chemical compounds on mammalian reproductive system, water-soluble fraction (WSF) extracted from heavy oil was administrated to mice for 2 weeks. WSF-administrated mice were crossed with either WSF- or distilled water-administrated group for mating experiment. When WSF-administrated male mice were used as a father, it reduced not only mating ratio, but also neonatal male ratio. The numbers of sperms of WSF-administrated male mice were decreased. In addition, abnormality of sperms such as bent or twisted tail was increased approximately 6-fold by WSF intake. The level of testosterone in serum from WSF-administrated mice was lower than that from control mice. Testosterone is the most important for the spermatogenesis in vertebrate. It is supposed from these findings, the decrease in the number of sperms may relate with the reduction of sex hormone level in serum. It is suggested from these results that the chemical substances in WSF affected the sperm function in reproductive system of male mice.
Assuntos
Poluentes Ambientais/toxicidade , Petróleo/toxicidade , Comportamento Sexual Animal/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Administração Oral , Animais , Poluentes Ambientais/química , Feminino , Masculino , Camundongos , Camundongos Endogâmicos ICR , Razão de Masculinidade , Solubilidade , Motilidade dos Espermatozoides/efeitos dos fármacos , Cauda do Espermatozoide/efeitos dos fármacos , Cauda do Espermatozoide/patologia , Espermatozoides/patologia , Testosterona/sangueRESUMO
The water-soluble fraction of kale (Brassica oleracea L. var. acephala DC.) had immunoglobulin (Ig) production stimulating activity in human hybridoma HB4C5 cells and human peripheral blood lymphocytes. The biochemical and physical properties of the main active substance in kale were found to be a heat-stable protein with a molecular weight higher than 50 kDa. The Ig production-stimulating factors were assumed to act on the translational and/or secreting processes of Igs. This Ig production-stimulating effect was also observed in lymphocytes from the mesenteric lymph node and Peyer's patches of mice that had been administered with the kale extract for 14 d. The partially purified kale extract was analyzed by LC-ESI-MS/MS, the result indicating ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) as an active substance. Rubisco from spinach indeed exhibited Ig production-stimulating activity in HB4C5 cells. These findings provide another beneficial aspect of kale as a health-promoting foodstuff.
Assuntos
Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/farmacologia , Brassica/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Água/química , Sequência de Aminoácidos , Animais , Citocinas/biossíntese , Feminino , Humanos , Hibridomas/citologia , Imunoglobulinas/biossíntese , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Linfócitos/metabolismo , Camundongos , Dados de Sequência Molecular , Solubilidade , Transcrição Gênica/efeitos dos fármacosRESUMO
Epithelial sodium channel (ENaC) is a heteromultimeric Na(+) channel at the apical membrane in the kidney, colon, and lung. Because ENaC plays a crucial role in regulating Na(+) absorption and extracellular fluid volume, its dysregulation causes severe phenotypes including hypertension, hypokalemia, and airway obstruction. Despite the importance of ENaC, its protein quality control mechanism remains less established. Here we firstly show the role of calreticulin (CRT), a lectin-like molecular chaperone in the endoplasmic reticulum (ER), on the regulation of ENaC. Overexpression and knockdown analyses clearly indicated that CRT positively affects the expression of each ENaC subunit (alpha, beta and gamma). CRT overexpression also up-regulated the cell surface expression of alpha-, beta- and gamma-ENaC. Moreover, we found that CRT directly interacts with each ENaC subunit. Although CRT knockdown did not affect the de novo synthesis of ENaC subunits, CRT overexpression decreased alpha-, beta- and gamma-ENaC expression in the detergent (RIPA)-insoluble fraction, suggesting that CRT enhanced the solubility of ENaC subunits. Consistent with the increased intracellular and cell surface expression of ENaC subunits, increased channel activity of ENaC was also observed upon overexpression of CRT. Our study thus identifies CRT as an ER chaperone that regulates ENaC expression and function.
Assuntos
Calreticulina/farmacologia , Canais Epiteliais de Sódio/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Animais , Células CHO , Calreticulina/genética , Cricetinae , Cricetulus , DNA Complementar , Retículo Endoplasmático , Canais Epiteliais de Sódio/metabolismo , Canais Epiteliais de Sódio/fisiologia , Chaperonas Moleculares , Ligação Proteica , Subunidades Proteicas , TransfecçãoRESUMO
Sorghum, Sorghum bicolor (L.) Moench, is the fifth most important cereal crop in the world. In this study, we identified the IgE production-suppressing activity of white sorghum bran extracts. White sorghum is one of the genotypes of sorghum. White sorghum bran extracts in 10 mM sodium phosphate buffer (pH 7.4) suppressed IgE production in human myeloma cell line U266. The extracts suppressed IgE production by decreasing mRNA transcription level of IgE, but they did not affect IgA or IgG production of mice splenocytes in vitro. Heat treatment and trypsin digestion did not affect IgE production-suppressing activity. The white sorghum bran extracts were fractionated by ultrafiltration, and the molecular weight of the active substance was estimated to be less than 1,000.
Assuntos
Imunoglobulina E/biossíntese , Mieloma Múltiplo/metabolismo , Extratos Vegetais/farmacologia , Sorghum/química , Animais , Sequência de Bases , Linhagem Celular Tumoral , Primers do DNA , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina E/genética , Camundongos , Mieloma Múltiplo/patologia , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Baço/efeitos dos fármacos , Baço/metabolismoRESUMO
We have previously demonstrated that collagen from jellyfish simulated immunoglobulin and cytokine production by human-human hybridoma line HB4C5 cells and by human peripheral blood lymphocytes (hPBL). The mode of action of the collagen as an immunostimulatory factor was investigated. The expression levels of immunoglobulin mRNAs in HB4C5 cells, and those of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, and transforming growth factor (TGF)-beta in hPBL were up-regulated by jellyfish collagen. In addition, this collagen activated IgM production by transcription-suppressed HB4C5 cells that had been treated with actinomycin D. This collagen also enhanced IgM production by translation-suppressed HB4C5 cells that had been treated with sodium fluoride, but was ineffective in accelerating IgM production by HB4C5 cells treated with cycloheximide. Moreover, the intracellular IgM level in HB4C5 cells treated with the post-translation inhibitor, monensin, was increased by this collagen. These results suggest that collagen from jellyfish stimulated not only the transcription activity, but also the translation activity for enhanced immunoglobulin and cytokine production.
Assuntos
Adjuvantes Imunológicos/farmacologia , Colágeno/imunologia , Cifozoários/química , Adjuvantes Imunológicos/isolamento & purificação , Animais , Bovinos , Linhagem Celular , Colágeno/isolamento & purificação , Citocinas/genética , Humanos , Imunoglobulina M/genética , Biossíntese de Proteínas/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transcrição Gênica/efeitos dos fármacosRESUMO
The 7',8'-stereochemistry of the tetrahydropyran sesquineolignans morinols A and B was determined as threo via synthetic studies and by comparison of NMR data of 7',8'-threo-morinol and 7',8'-erythro-morinol. This study also confirmed that the biosynthetic process produces enantiomeric mixtures of morinols A and B. This was ascertained by comparing the specific rotations of synthesized morinols A and B with those of naturally occurring morinols A and B.
Assuntos
Lignanas/química , Magnoliopsida/química , Plantas Medicinais/química , Piranos/química , Lignanas/síntese química , Lignanas/isolamento & purificação , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Piranos/síntese química , Piranos/isolamento & purificaçãoRESUMO
Certain edible large jellyfishes belonging to the order Rhizostomeae are consumed in large quantities in China and Japan. The exumbrella part of the edible jellyfish Stomolophus nomurai was cut and soaked in dilute hydrochloric acid solution (pH 3.0) for 12 h, and heated at 121 degrees C for 20 min. The immunostimulation effects of the jellyfish extract were examined. The jellyfish extract enhanced IgM production of human hybridoma HB4C5 cells 34-fold. IgM and IgG production of human peripheral blood lymphocytes (PBL) were also accelerated, 2.8- and 1.4-fold respectively. Moreover, production of interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha by human PBL was stimulated 100- and 17-fold respectively. Collagenase treatment inactivated the immunostimulation activity of the jellyfish extract. In addition, purified collagen from bovine Achilles' tendon accelerated IgM production of hybridoma cells. These facts mean that collagen has an immunostimulation effect, and that the active substance in jellyfish extract is collagen.