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1.
Insect Biochem Mol Biol ; 116: 103261, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31698082

RESUMO

A network of serine proteases (SPs) and their non-catalytic homologs (SPHs) activates prophenoloxidase (proPO), Toll pathway, and other insect immune responses. However, integration and conservation of the network and its control mechanisms have not yet been fully understood. Here we present evidence that these responses are initiated through a conserved serine protease and negatively regulated by serpins in two species, Manduca sexta and Anopheles gambiae. We have shown that M. sexta serpin-12 reduces the proteolytic activation of HP6, HP8, proPO activating proteases (PAPs), SPHs, and POs in larval hemolymph, and we hypothesized that these effects are due to the inhibition of the immune pathway-initiating protease HP14. To test whether these changes are due to HP14 inhibition, we isolated a covalent complex of HP14 with serpin-12 from plasma using polyclonal antibodies against the HP14 protease domain or against serpin-12, and confirmed formation of the complex by 2D-electrophoresis, immunoblotting, and mass spectrometry. Upon recognition of bacterial peptidoglycans or fungal ß-1,3-glucan, the zymogen proHP14 became active HP14, which formed an SDS-stable complex with serpin-12 in vitro. Activation of proHP21 by HP14 was suppressed by serpin-12, consistent with the decrease in steps downstream of HP21, proteolytic activation of proPAP3, proSPH1/2 and proPO in hemolymph. Guided by the results of phylogenetic analysis, we cloned and expressed A. gambiae proSP217 (an ortholog of HP14) and core domains of A. gambiae serpin-11 and -17. The recombinant SP217 zymogen became active during expression, with cleavage between Tyr394 and Ile395. Both MsHP14 and AgSP217 cleaved MsSerpin-12 and AgSRPN11 at Leu*Ser (P1*P1') and formed complexes in vitro. ProPO activation in M. sexta plasma increased after recombinant AgSP217 had been added, indicating that it may function in a similar manner as the endogenous initiating protease HP14. Based on these data, we propose that inhibition of an initiating modular protease by a serpin may be a common mechanism in holometabolous insects to regulate proPO activation and other protease-induced immune responses.


Assuntos
Anopheles/imunologia , Manduca/imunologia , Serpinas/metabolismo , Animais , Anopheles/metabolismo , Catecol Oxidase/genética , Catecol Oxidase/metabolismo , Precursores Enzimáticos/genética , Precursores Enzimáticos/metabolismo , Hemolinfa/enzimologia , Proteínas de Insetos/metabolismo , Larva/genética , Larva/imunologia , Larva/metabolismo , Manduca/genética , Manduca/metabolismo , Peptidoglicano/farmacologia , Filogenia , Serina Proteases/genética , Serina Proteases/metabolismo , beta-Glucanas/farmacologia
2.
Molecules ; 24(19)2019 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-31590413

RESUMO

BACKGROUND: Inflammatory bowel diseases are an important health problem. Therefore, the aim of the present study was to compare the impact of isolated oat beta-glucan fractions of low and high molecular weight, taken as dietary supplementation, on inflammatory markers in the colitis model. METHODS: Two groups of Sprague-Dawley rats-control and with experimentally induced colitis-were subsequently divided into three subgroups and fed over 21 days feed supplemented with 1% of low (ßGl) or high (ßGh) molecular weight oat beta-glucan fraction or feed without supplementation. The level of colon inflammatory markers, cytokines, and their receptors' genes expressions and immune cells numbers were measured by ELISA, RT-PCR, and by flow cytometry methods, respectively. RESULTS: The results showed moderate inflammation affecting the colon mucosa and submucosa, with significant changes in the number of lymphocytes in the colon tissue, elevated cytokines and eicosanoid levels, as well as disruption of the main cytokine and chemokine cell signaling pathways in colitis rats. Beta-glucans supplementation caused a reverse in the percentage of lymphocytes with stronger effects of ßGh and reduction of the levels of the inflammatory markers, and improvement of cytokine and chemokine signaling pathways with stronger effects of ßGl supplementation. CONCLUSIONS: The results indicate the therapeutic effect of dietary oat beta-glucan supplementation in the colitis in evident relation to the molecular weight of polymer.


Assuntos
Anti-Inflamatórios/administração & dosagem , Avena/química , Colite/dietoterapia , Ácido Trinitrobenzenossulfônico/efeitos adversos , beta-Glucanas/administração & dosagem , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Colite/induzido quimicamente , Colite/genética , Colite/imunologia , Citocinas/genética , Citocinas/metabolismo , Suplementos Nutricionais , Modelos Animais de Doenças , Regulação da Expressão Gênica/efeitos dos fármacos , Contagem de Linfócitos , Masculino , Peso Molecular , Ratos , Ratos Sprague-Dawley , beta-Glucanas/química , beta-Glucanas/farmacologia
3.
J Med Food ; 22(10): 1009-1021, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31536448

RESUMO

Constipation is a common gastrointestinal disorder characterized by changes in intestinal habits. Increasing evidence indicates that long-term use of irritant laxatives causes serious side effects. Meanwhile, more than 50% of patients are dissatisfied with sense of use of non-prescriptional laxatives. ß-glucans are natural polysaccharides widely found in yeast, fungus, and plants, which have been reported to exhibit various pharmacological effects. The aim of this study was to characterize the effect of ß-glucans extracted from the bread yeast cell wall on loperamide-induced constipation mice. Forty mice were fed with loperamide (10 mg/kg) to make the constipation model and a diet supplemented with 2.5, 5, and 10 mg/kg ß-glucan. We assessed the defecation frequency, intestinal transit function of mice, as well as used high-throughput sequencing to analyze the intestinal microbiota composition and functional biological profiles data. Meanwhile, we detected expression of neurotransmitters including acetylcholinesterase, substance P, and serotonin (5-HT) and expression of tight junction protein (TJP) including zonula occludens-1 and mucin-2 in distal colon to characterize the possible molecular mechanisms. ß-glucans significantly enhanced intestinal motility and provided a possibility to regulate the expression of neurotransmitters and TJP in mice. The intestinal microecological portion of the treatment group partially recovered and was closer to the normal group. This study showed that ß-glucans can influence the intestinal microbiota and restore microecological balance to regulate the express of neurotransmitters and TJP to recover intestinal epithelial mechanical barrier. We suggested that ß-glucans could be used as an active nutritional supplement to protect the damaged intestinal barrier and help patients who have constipation complications and dysbiosis.


Assuntos
Constipação Intestinal/tratamento farmacológico , Microbioma Gastrointestinal , Loperamida/efeitos adversos , Saccharomyces cerevisiae/química , beta-Glucanas/farmacologia , Acetilcolinesterase/metabolismo , Animais , Constipação Intestinal/induzido quimicamente , Defecação/efeitos dos fármacos , Trânsito Gastrointestinal/efeitos dos fármacos , Masculino , Metagenoma , Camundongos , Camundongos Endogâmicos BALB C , Mucina-2/metabolismo , Serotonina/metabolismo , Substância P/metabolismo , Proteína da Zônula de Oclusão-1/metabolismo
4.
J Microbiol Biotechnol ; 29(11): 1693-1706, 2019 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-31546298

RESUMO

Atopic dermatitis (AD) is a chronic inflammatory skin disease of mainly infants and children. Currently, the development of safe and effective treatments for AD is urgently required. The present study was conducted to investigate the immunomodulatory effects of yeast-extracted ß-1,3/1,6-glucan and/or Lactobacillus plantarum (L. plantarum) LM1004 against AD-like symptoms. To purpose, ß-1,3/1,6-glucan and/or L. plantarum LM1004 were orally administered to AD-induced animal models of rat (histamine-induced vasodilation) and mouse (pruritus and contact dermatitis) exhibiting different symptoms of AD. We then investigated the treatment effects on AD-like symptoms, gene expression of immune-related factors, and gut microbiomes. Oral administration of ß-1,3/1,6-glucan (0.01 g/kg initial body weight) and/or 2 × 1012 cells/g L. plantarum LM1004 (0.01 g/kg initial body weight) to ADinduced animal models showed significantly reduced vasodilation in the rat model, and pruritus, edema, and serum histamine in the mouse models (p < 0.05). Interestingly, ß-1,3/1,6- glucan and/or L. plantarum LM1004 significantly decreased the mRNA levels of Th2 and Th17 cell transcription factors, while the transcription factors of Th1 and Treg cells, galactin-9, filaggrin increased, which are indicative of enhanced immunomodulation (p < 0.05). Moreover, in rats with no AD induction, the same treatments significantly increased the relative abundance of phylum Bacteroidetes and the genus Bacteroides. Furthermore, bacterial taxa associated with butyrate production such as, Lachnospiraceae and Ruminococcaceae at family, and Roseburia at genus level were increased in the treated groups. These findings suggest that the dietary supplementation of ß-1,3/1,6-glucan and/or L. plantarum LM1004 has a great potential for treatment of AD as well as obesity in humans through mechanisms that might involve modulation of host immune systems and gut microbiota.


Assuntos
Dermatite Atópica/terapia , Fatores Imunológicos/administração & dosagem , Lactobacillus plantarum , Probióticos/administração & dosagem , beta-Glucanas/administração & dosagem , Administração Oral , Animais , Citocinas/genética , Dermatite Atópica/induzido quimicamente , Dermatite Atópica/patologia , Modelos Animais de Doenças , Microbioma Gastrointestinal/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Masculino , Camundongos , Probióticos/farmacologia , Ratos Sprague-Dawley , Pele/efeitos dos fármacos , Pele/patologia , Linfócitos T/imunologia , Fatores de Transcrição/genética , Resultado do Tratamento , beta-Glucanas/farmacologia
5.
Nutrients ; 11(9)2019 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-31470503

RESUMO

Chronic obstructive pulmonary disease (COPD), a lung disease caused by chronic exposure to cigarette smoke, increases the number of inflammatory cells such as macrophages and neutrophils and emphysema. Isoflavone is a polyphenolic compound that exists in soybeans. Daidzein and genistein, two types of isoflavones, have been reported to have anti-inflammatory effects in various organs. We hypothesized that the daidzein-rich soy isoflavone aglycones (DRIAs) attenuate cigarette smoke-induced emphysema in mice. Mice were divided into four groups: the (i) control group, (ii) isoflavone group, (iii) smoking group, and (iv) isoflavone + smoking group. The number of inflammatory cells in the bronchoalveolar lavage fluid (BALF) and the airspace enlargement using the mean linear intercept (MLI) were determined 12 weeks after smoking exposure. Expressions of neutrophilic inflammatory cytokines and chemokines were also examined. In the isoflavone + smoking group, the number of neutrophils in BALF and MLI was significantly less than that in the smoking group. Furthermore, the gene-expressions of TNF-α and CXCL2 (MIP-2) in the isoflavone + smoking group were significantly less than those in the smoking group. Supplementation of the COPD murine model with DRIAs significantly attenuates pathological changes of COPD via suppression of neutrophilic inflammation.


Assuntos
Anti-Inflamatórios/farmacologia , Isoflavonas/farmacologia , Pulmão/efeitos dos fármacos , Infiltração de Neutrófilos/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Pneumonia/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Enfisema Pulmonar/tratamento farmacológico , Fumaça , Produtos do Tabaco , beta-Glucanas/farmacologia , Animais , Citocinas/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Mediadores da Inflamação/imunologia , Mediadores da Inflamação/metabolismo , Pulmão/imunologia , Pulmão/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Neutrófilos/imunologia , Neutrófilos/metabolismo , Pneumonia/etiologia , Pneumonia/imunologia , Pneumonia/metabolismo , Doença Pulmonar Obstrutiva Crônica/etiologia , Doença Pulmonar Obstrutiva Crônica/imunologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , Enfisema Pulmonar/etiologia , Enfisema Pulmonar/imunologia , Enfisema Pulmonar/metabolismo , Transdução de Sinais
6.
Fish Shellfish Immunol ; 94: 427-433, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31536766

RESUMO

Our study is considered to attempt reducing the immune-toxic and antioxidant impacts of exposure to fipronil (FP) on Nile tilapia, Oreochromis niloticus using the ß-glucan (ßG). Two hundred and seventy fingerlings of Nile tilapia were divided randomly into six groups (45 tilapias of each, in 3 replicates): group I control (CT) group nourished on a basal diet. Group II (ßG) nourished a basal diet supplemented with 0.4% ßG. Groups III (1/20 FP) and V (1/10 FP) was exposed to 1/20 and 1/10 of the 96 h LC50 of FP in water and nourished the basal diet respectively. Groups IV (1/20 FP+ ßG) and VI (1/10 FP+ ßG) were exposed to 1/20 and 1/10 FP concomitantly with 0.4% ßG supplementation for 90 successive days. Growth performance metrics were higher in ßG group than CT. While those metrics were fallen at exposure to 1/20 or 1/10 FP. Supplementation with ßG elevated the IgM and lysozyme levels.Whereas, tilapias exposed to FP only at different concentration showed lowering of those compared to CT. Supplementation with ßG was effectively augmented IgM and lysozyme in 1/20 FP exposed tilapias. Furthermore, in a minor grade at 1/10 FP exposed tilapias. Exposure to FP increased the activities of hepatic markers chiefly at 1/10, however the ßG supplementation was successfully improved these markers. There was imbalance of cortisol level at FP exposure where, ßG combining to FP alleviate this disparity. There was fallen in LDH, MDH and FDPase in ßG tilapias where continuing raise in 1/10 FP followed by 1/20 FP. ßG supplementation raise the level of GSH, without significant variations in MDA conversely occurs in FP alone. Genes expression of ßG caused raise of both GPx and GR, without fluctuations in CAT and SOD. Exposure to FP diminishes all evaluated antioxidant genes. It could fulfilled that supplementation with ßG successfully alleviated the immune-toxic and antioxidant impact of FP in tilapias.


Assuntos
Adjuvantes Imunológicos/farmacologia , Antioxidantes/metabolismo , Ciclídeos/imunologia , Expressão Gênica/efeitos dos fármacos , Inseticidas/efeitos adversos , Pirazóis/efeitos adversos , beta-Glucanas/farmacologia , Ração Animal/análise , Animais , Ciclídeos/genética , Ciclídeos/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga
7.
Fish Shellfish Immunol ; 93: 732-742, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31415901

RESUMO

This study evaluates the effect of dietary supplementation of immunostimulants on the Chinese mitten crab (Eriocheir sinensis) with a single administration of mannan oligosaccharide (MOS), or its combination with either ß-glucan or with inulin for 8 weeks. Four diets included an untreated control diet (C), MOS alone (3 g kg-1, M), MOS with ß-glucan (3 g kg -1 MOS + 1.5 g kg -1 ß-glucan, MB), and MOS with inulin (3 g kg -1 MOS + 10 g kg -1 inulin, MI). The weight gain and specific growth rate of the crabs fed M, MB, and MI diets were improved by lowing feed conversion ratio. The growth and feed utilization of the crabs fed the MB diet were improved compared with the other three groups. The crabs fed the M, MB and MI diets showed a higher intestinal trypsin activity than that in the M and control groups. The highest trypsin activity in the hepatopancreas was observed in the MB group. Crabs fed M, MB and MI diets increased antioxidant system-related enzyme activities, but reduced malondialdehyde. The highest activities of alkaline phosphatase, acid phosphatase, lysozyme and phenol oxidase in the gut and the respiratory burst of the crabs were found in the MB group. The MB diet promoted the mRNA expression of E. sinensis immune genes (ES-PT, ES-Relish, ES-LITAF, p38MAPK and Crustin) compared with the control. After 3 days of infection with Aeromonas hydrophila, the highest survival of crabs was also found in the MB group. This study indicates that the combination of MOS with ß-glucan or with inulin can improve growth, antioxidant capacity, non-specific immunity and disease resistance in E. sinensis.


Assuntos
Adjuvantes Imunológicos/farmacologia , Braquiúros/imunologia , Imunidade Inata/efeitos dos fármacos , Inulina/farmacologia , Mananas/farmacologia , Oligossacarídeos/farmacologia , beta-Glucanas/farmacologia , Aeromonas hydrophila/fisiologia , Animais , Antioxidantes/metabolismo , Braquiúros/crescimento & desenvolvimento , Braquiúros/metabolismo , Braquiúros/microbiologia , Distribuição Aleatória
8.
Fish Shellfish Immunol ; 94: 10-16, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31465869

RESUMO

In crustaceans, it has been suggested that specific protection against pathogens could be triggered by vaccines and biological response modifiers; although the specific mechanisms of this protection have not been clarified yet. In the crayfish Cherax quadricarinatus, a humoral lectin (CqL) binds its own granular hemocytes through a specific receptor (CqLR) and increases the production of reactive oxygen species (ROS). In the present study, we challenged in vivo crayfishes with immunostimulants, ß-glucan (200 µg/kg) or LPS (20 µg/kg), and identified the participation of cellular and humoral mechanisms. The stimulants generated a complex modification in the total hemocytes count (THC), as well as in the proportion of hemocyte subsets. At 2 h after the challenge, the largest value in THC was observed in either challenged crayfishes. Furthermore, at the same time, hyaline hemocytes were the most abundant subset in the hemolymph; after 6 h, granular hemocytes (GH) were the most abundant hemocyte subset. It has been observed that a specific subset of GH possesses a CqLR that has been related to ROS production. After 2 and 6 h of the ß-glucan challenge, a significant increase in CqLR expression was observed in the three circulating hemocyte subsets; also, an increased expression of CqL was detected in a granular hemocytes sub-population. After 2 and 6 h of stimulation, the specific activity of the serum lectin challenged with ß-glucan was 250% and 160% higher than in the LPS-treated-group, respectively (P < 0.05). Hemocytes from challenged crayfishes were stimulated ex vivo with CqL, ROS production was 180% higher in hemocytes treated with ß-glucan + CqL than in hemocytes treated with LPS + CqL (P < 0.05). The results evidence the effectivity of immune stimulators to activate specific crayfish defense mechanisms, the participation of CqL and its receptor (CqLR) could play an important role in the regulation of immune cellular functions, like ROS production, in Cherax quadricarinatus.


Assuntos
Astacoidea/genética , Astacoidea/imunologia , Expressão Gênica/efeitos dos fármacos , Imunidade Celular/genética , Imunidade Humoral/genética , Lectinas/genética , Receptores Mitogênicos/genética , Adjuvantes Imunológicos/farmacologia , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Astacoidea/efeitos dos fármacos , Hemócitos , Hemolinfa/metabolismo , Lectinas/metabolismo , Lipopolissacarídeos/farmacologia , Receptores Mitogênicos/metabolismo , beta-Glucanas/farmacologia
9.
J Agric Food Chem ; 67(32): 9070-9078, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31343168

RESUMO

In this study, an immunologically active novel microparticulate mushroom ß-glucan (PRA-1p) was prepared using an alkali-soluble glucan PRA-1 by an emulsification and cross-linking method. PRA-1 was a hyperbranched (1→3),(1→6)-ß-d-glucan with a degree of branching of 0.89, isolated from the sclerotia of Polyporus rhinocerus. PRA-1 had a rod-like conformation, while PRA-1p exhibited a monodisperse and homogeneous spherical conformation with a diameter ranging from 0.3 to 2.0 µm in water. PRA-1p significantly induced nitric oxide and reactive oxygen species production as well as morphological changes of murine macrophages (RAW 264.7 cells) and upregulated their phagocytic activity. Furthermore, PRA-1p treatment markedly enhanced the secretion of cytokines, including cutaneous T cell-attracting chemokine 27, granulocyte-colony-stimulating factor, monocyte chemoattractant protein 1, macrophage inflammatory protein 1α, macrophage inflammatory protein 2, regulated on activation, normal T cell expressed and secreted, soluble tumor necrosis factor receptor 1, and tissue inhibitors of metalloproteinases. Activation of RAW 264.7 cells triggered by PRA-1p was associated with activation of inducible nitric oxide synthase, nuclear factor κB, extracellular signal-regulated kinase, and protein kinase B. This work suggests that novel PRA-1p derived from the mushroom sclerotia of P. rhinocerus has potential application as an immunostimulatory agent.


Assuntos
Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Polyporus/química , beta-Glucanas/química , beta-Glucanas/farmacologia , Animais , Quimiocina CCL27/genética , Quimiocina CCL27/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Fatores Imunológicos/isolamento & purificação , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos , NF-kappa B/genética , NF-kappa B/imunologia , Óxido Nítrico/imunologia , Fagocitose/efeitos dos fármacos , Extratos Vegetais/isolamento & purificação , Polyporus/imunologia , Células RAW 264.7 , Espécies Reativas de Oxigênio/imunologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , beta-Glucanas/isolamento & purificação
10.
Fish Shellfish Immunol ; 93: 1076-1083, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31352115

RESUMO

In this study, we show that ß-glucan can modulate cortisol release in fish. We simulated a common situation in aquaculture: the transport of fish followed by contact with an opportunistic pathogen and observed what effect glucan had on the immune and stress response in these conditions. Pacu (Piaractus mesopotamicus) were fed with a diet containing ß-glucan (0.1%) for 15 days prior to transport followed by an injection with heat-killed Aeromonas hydrophila. We sampled fish before transport, at arrival and at 3 and 24 h after bacterial injection. ß-Glucans are used in aquaculture and have a known immunostimulatory effect, which was observed in this study. The results showed that ß-glucan modulated the plasma cortisol levels differently by increasing these levels up to 24 h after transport and preventing the increase caused by bacterial inoculum injection. In addition, ß-glucan enhanced the activity of the complement system at 24 h and reduced the monocytes and lymphocytes number in peripheral blood at 3 and 24 h after bacterial inoculation. Our results suggest that ß-glucan modulated a bidirectional interaction between the stress and the immune responses. The modulation of cortisol levels and the immunostimulation by ß-glucan at different moments in our study suggest the compound has a protective effect by avoiding higher levels of the hormone and improving resistance against bacterial infection in pacu. These results add evidence to support the use of ß-glucan as an immunomodulator in the aquaculture industry.


Assuntos
Adjuvantes Imunológicos/farmacologia , Caraciformes/imunologia , Doenças dos Peixes/imunologia , Hidrocortisona/sangue , Imunidade Inata/efeitos dos fármacos , beta-Glucanas/farmacologia , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Dieta/veterinária , Suplementos Nutricionais/análise , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Distribuição Aleatória , Estresse Fisiológico
11.
Carbohydr Polym ; 222: 114996, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31320070

RESUMO

H6PC20, a water-soluble ß-D-glucan, with a molecular mass of 2390 kDa (HPSEC-MALLS-RI), was obtained by extracting the fresh fruiting bodies of Hericium erinaceus at mature stage with hot water, freeze-thawing, precipitating and further rinsing with 20% (v/v) of ethanol. The primary structure of H6PC20 was proposed as a ß-(1→3)-linked-D-glucan with a (1→6)-ß-linked-D- glucopyranosyl side-branching unit on every third residue by using methylation analysis, 1H and 13C NMR spectroscopy, including COSY, TOCSY, HSQC and HMBC. The ρ value (Rg/Rh) and the exponent (α) of [η] = k Mwα of H6PC20 were calculated as 1.86 and 0.54, respectively, indicating flexible random coil conformation of H6PC20 in aqueous solution. H6PC20 could induce the proliferation of lymphocytes and enhance the expression of inflammatory cytokines IL-6, TNF-α and IL-1ß secreted by THP-1 macrophages. The present study indicated that the macromolecular ß-glucan H6PC20 possessed potent immune-enhancing bioactivities and could be a potential ingredient for healthy food or medicine development.


Assuntos
Agaricales/química , Fatores Imunológicos/farmacologia , beta-Glucanas/farmacologia , Animais , Sequência de Carboidratos , Citocinas/metabolismo , Carpóforos/química , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/isolamento & purificação , Linfócitos/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Peso Molecular , Células THP-1 , beta-Glucanas/química , beta-Glucanas/isolamento & purificação
12.
Carbohydr Polym ; 222: 114990, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31320072

RESUMO

A water insoluble ß-glucan (PS), with molecular mass ∼9.16 × 104 Da was isolated from the 4% alkaline extract of an edible mushroom, Pleurotus djamor and found to consist of (1→3)-ß-d-glucopyranosyl moiety. The structure of the PS was elucidated on the basis of total hydrolysis, methylation analysis, periodate oxidation, and NMR experiments (1H, 13C, DQF-COSY, DEPT-135, and HSQC). The structure of the repeating unit of the polysaccharide was established as: →3)-ß-d-Glcp-(1→. The water insoluble ß-glucan showed cytotoxic effect against PA1 cells, where˜50% population was destroyed at 100 µg/mL concentration, and almost all cells at 250 µg/mL concentration. The wound healing assay showed significant anticarcinogenic effect against ovarian carcinoma PA1 cells after 48 h of treatment.


Assuntos
Antineoplásicos/farmacologia , Neoplasias Ovarianas/tratamento farmacológico , Pleurotus/metabolismo , beta-Glucanas , Sequência de Carboidratos , Feminino , Humanos , Solubilidade , Células Tumorais Cultivadas , beta-Glucanas/química , beta-Glucanas/farmacologia
13.
PLoS One ; 14(6): e0218118, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31185060

RESUMO

We investigated whether supplementation with the barley line BARLEYmax (Tantangara; BM), which contains three fermentable fibers (fructan, ß-glucan, and resistant starch), modifies the microbiota in cecal and distal colonic digesta in addition to short-chain fatty acids (SCFAs) production more favorably than supplementation with a high-ß-glucan barley line (BG012; BG). Male Sprague-Dawley rats were randomly divided into 3 groups that were fed an AIN-93G-based diet that contained 5% fiber provided by cellulose (control), BM or BG. Four weeks after starting the respective diets, the animals were sacrificed and digesta from the cecum, proximal colon and distal colon were collected and the SCFA concentrations were quantified. Microbiota in the cecal and distal colonic digesta were analyzed by 16S rRNA sequencing. The concentrations of acetate and n-butyrate in cecal digesta were significantly higher in the BM and BG groups than in the control group, whereas the concentration of total SCFAs in cecal digesta was significantly higher only in the BM group than in the control group. The concentrations of acetate and total SCFAs in the distal colonic digesta were significantly higher only in the BM group than in the control group. The abundance of Bacteroidetes in cecal digesta was significantly higher in the BM group than in the control group. In contrast, the abundance of Firmicutes in cecal digesta was significantly lower in the BM and BG groups than in the control group. These results indicated that BM increased the concentration of total SCFAs in the distal colonic digesta. These changes might have been caused by fructan and resistant starch in addition to ß-glucan. In conclusion, fermentable fibers in BM reached the distal colon and modified the microbiota, leading to an increase in the concentration of total SCFAs in the distal colonic digesta, more effectively compared with the high-ß-glucan barley line (BG).


Assuntos
Bacteroidetes/metabolismo , Ceco/microbiologia , Colo/microbiologia , Ácidos Graxos/biossíntese , Hordeum , Microbiota/efeitos dos fármacos , beta-Glucanas/farmacologia , Ração Animal , Animais , Ratos , Ratos Sprague-Dawley
14.
Int J Biol Macromol ; 136: 1169-1175, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31170489

RESUMO

Gemcitabine (2'-deoxy-2',2'-difluorocytidine, dFdC) is one of the most effective chemotherapy drugs commonly used for treatment of various tumors. Despite its significant anticancer effects, some adverse effects create obstacles to treatment. The main toxicity of gemcitabine is myelosuppression, which not only reduces patient quality of life, but also hinders further anticancer treatment. In this respect, immunotherapy can address these drawbacks because of its ability to enhance the patient's immune system. To improve immune system function, yeast-derived ß-glucans, which are well-known biologic response modifiers, were administered to gemcitabine-treated mice. The in vivo experiment revealed that orally administered yeast (1 → 3)-(1 → 6)-ß-d-glucan effectively alleviated myelosuppression associated with gemcitabine-induced pancytopenia. Moreover, analysis of myelopoiesis-related cytokine expression through real-time PCR demonstrated that ß-glucan treatment significantly upregulated hematopoietic responses in gemcitabine-treated mice. Furthermore, orally administered ß-glucan significantly induced the expression of IFN-γ and IL-2 in splenocytes of gemcitabine-treated mice. It also restored the cytotoxicity of splenocytes against YAC-1 in gemcitabine-treated mice and displayed a positive effect on gemcitabine-damaged bone marrow tissue. In conclusion, yeast ß-glucans have the potential to be used as adjuvants for alleviating chemotherapy-induced immunosuppression in patients.


Assuntos
Desoxicitidina/análogos & derivados , Imunossupressão/efeitos adversos , Leveduras/química , beta-Glucanas/farmacologia , Animais , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/patologia , Linhagem Celular Tumoral , Citocinas/metabolismo , Desoxicitidina/efeitos adversos , Desoxicitidina/antagonistas & inibidores , Hematopoese/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pancitopenia/induzido quimicamente , Pancitopenia/tratamento farmacológico , Pancitopenia/imunologia , Baço/efeitos dos fármacos , Baço/imunologia , beta-Glucanas/uso terapêutico
15.
Int J Mol Sci ; 20(9)2019 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-31083511

RESUMO

Exocellular (1→6)-ß-d-glucan (lasiodiplodan) produced by the fungus Lasiodiplodia theobromae MMPI was derivatized by carboxymethylation using different concentrations of a derivatizing agent. Lasiodiplodan was derivatized by carboxymethylation in an attempt to increase its solubility and enhance its biological activities. Carboxymethylglucans with degrees of substitution (DS) of 0.32, 0.47, 0.51, 0.58, and 0.68 were produced and characterized. FTIR analysis showed a band of strong intensity at 1600 cm-1 and an absorption band at 1421 cm-1, resulting from asymmetric and symmetrical stretching vibrations, respectively, of the carboxymethyl group COO- in the carboxymethylated samples. Thermal analysis showed that native lasiodiplodan (LN) and carboxymethylated derivatives (LC) exhibited thermal stability up to 200-210 °C. X-ray diffractometry demonstrated that both native and carboxymethylated lasiodiplodan presented predominantly an amorphous nature. Scanning electron microscopy revealed that carboxymethylation promoted morphological changes in the biopolymer and increased porosity, and alveolar structures were observed along the surface. The introduction of carboxymethyl groups in the macromolecule promoted increased solubility and potentiated the hydroxyl radical-scavenging activity, suggesting a correlation between degree of substitution and antioxidant activity.


Assuntos
Antioxidantes/farmacologia , Ascomicetos/química , beta-Glucanas/metabolismo , beta-Glucanas/farmacologia , Depuradores de Radicais Livres/química , Metilação , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura Ambiente , Termogravimetria , Difração de Raios X , beta-Glucanas/química
16.
Int J Biol Macromol ; 135: 619-629, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31132443

RESUMO

The aim of this study was to investigate the effects of biomacromolecules mannan/ß-glucans from yeast cell wall (BYCW) to alleviate Deoxynivalenol(DON)-induced injury. Considering that DON has strong oxidizing effect and stimulates autophagy and apoptosis, we examined the effects of BYCW on consequent oxidative stress damage indicators, cells autophagy and apoptosis induced by DON using the porcine jejunum epithelial cell lines (IPEC-J2) as a cell culture model. The results showed that application of BYCW could reverse the decrease of cell viability by DON significantly, and suppress the levels of tumor necrosis factor-α (TNF-α) and interleukin-8 and -6 (IL-8 and IL-6), except IL-1ß. Further experiments revealed that BYCW treatment counteracted the DON-induced down-regulation of intracellular glutathione (GSH) and up-regulation of reactive oxygen species (ROS) and malondialdehyde (MDA). Through western blot analysis, we observed that BYCW treatment was able to down-regulate the expression of autophagy protein LC3-II and up-regulate the expression of P62 protein against DON, which suggested that autophagy induced by DON may be suppressed. Altogether, these results indicated a potential ability of supplementation of BYCW to improve cell growth and metabolism as well as the preventive properties of BYCW against the DON-induced cell damage by activating antioxidant system.


Assuntos
Autofagia/efeitos dos fármacos , Parede Celular/química , Mananas/química , Estresse Oxidativo/efeitos dos fármacos , Tricotecenos/toxicidade , beta-Glucanas/química , beta-Glucanas/farmacologia , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citoproteção/efeitos dos fármacos , Glutationa/metabolismo , Malondialdeído/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Saccharomyces cerevisiae/citologia
17.
Fish Shellfish Immunol ; 91: 315-324, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31129185

RESUMO

An 8-week trial was conducted to evaluate the effect of dietary ß-glucan supplement (0, 0.01%, 0.02%, or 0.04%) on growth and health of Pacific white shrimp Litopenaeus vannamei at low salinity of 3 practical salinity unit (psu). The L. vannamei fed 0.02% and 0.04% ß-glucan gained more weight and showed higher activities of protease, amylase, superoxide dismutase, and glutathione peroxidase in the intestine than in the control (0% ß-glucan). The L. vannamei fed 0.04% ß-glucan had a higher condition factor than those fed the control diet. Amylase activity in the hepatopancreas of L. vannamei fed 0.02% ß-glucan was higher than those fed the control diet. Dietary ß-glucan supplement increased the mRNA expressions of Toll-like receptor, myostatin, immune deficiency or heat shock protein 70, but decreased the mRNA expressions of tumor necrosis factor-α and C-type lectin 3 in both hepatopancreas and intestine. The response of intestine microbiota in L. vannamei fed 0.04% ß-glucan was further compared to the control. The 0.04% ß-glucan supplement reduced richness and diversity of the intestinal microbial community as indicated by the low values of Chao1 estimator, ACE estimator, Simpson index and Shannon diversity index. Abundances of Bacillus, Chitinibacter, Geobacillus and Vibrio in the intestine increased, while Flavobacterium, Microbacterium and Mycobacterium decreased significantly in L. vannamei fed 0.04% ß-glucan compared to the control. This study indicates that dietary ß-glucan supplement at 0.02%-0.04% can significantly improve digestibility, antioxidant capacity and immunity in L. vannamei, and thus improve growth performance and survival at low salinity. These beneficial effects of ß-glucan probably are related to the dominance of probiotics over potential pathogens in the intestine.


Assuntos
Penaeidae/crescimento & desenvolvimento , Penaeidae/imunologia , Probióticos/farmacologia , Salinidade , beta-Glucanas/farmacologia , Ração Animal/análise , Animais , Dieta , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Penaeidae/metabolismo , Probióticos/administração & dosagem , Distribuição Aleatória , beta-Glucanas/administração & dosagem
18.
Molecules ; 24(8)2019 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-31022848

RESUMO

The purpose of this study is to develop a robust approach to obtain ß glucans from Lentinus edodes and to characterize their structural and biological properties for sustainable utilization. The alkali extraction was optimized with an orthogonal experimental design, and a concise process for obtaining specific targeting polysaccharides from Lentinus edodes was developed in this study. After purification with a Q-Sepharose Fast Flow strong anion-exchange column, the monosaccharide composition, a methylation analysis, and NMR spectroscopy were employed for their structural characterizations. LeP-N2 was found to be composed of (1→6)-ß-d-glucans with minor ß-(1→3) glucosidic side chains. Atomic force microscopy (AFM) and high-performance gel permeation chromatography-refractive index-multi-angle laser light scattering (HPGPC-RI-MALLS) also revealed LeP-N2 exhibiting a compact unit in aqueous solution. This (1→6)-ß-d-glucan was tested for antioxidant activities with IC50 at 157 µg/mL. Moreover, RAW 264.7 macrophage activation indicated that the release of nitric oxide (NO) and reactive oxygen species (ROS) was markedly increased with no cytotoxicity at a dose of 100 µg/mL. These findings suggest that the (1→6)-ß-d-glucans obtained from Lentinus edodes could serve as potential agents in the fields of functional foods or medicine.


Assuntos
Antioxidantes/química , Polissacarídeos/química , Cogumelos Shiitake/química , beta-Glucanas/química , Animais , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Proliferação de Células/efeitos dos fármacos , Cromatografia em Gel , Espectroscopia de Ressonância Magnética , Camundongos , Microscopia de Força Atômica , Estrutura Molecular , Polissacarídeos/isolamento & purificação , Células RAW 264.7 , Espécies Reativas de Oxigênio/química , Água/química , beta-Glucanas/isolamento & purificação , beta-Glucanas/farmacologia
19.
Fish Shellfish Immunol ; 89: 132-140, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30930276

RESUMO

Toll-like receptors (TLRs) are a category of most well recognized pattern recognition molecules that act on a vital role in both innate and adaptive immunity. In the present study, a novel toll-like receptor (McTLRw) was identified and characterized in thick shell mussel Mytilus coruscus. McTLRw possesses one intracellular Toll/interleukin-1 (IL-1) receptor (TIR) domain, one transmembrane region (TM), one leucine rich repeat N-terminal domain (LRR_NT) and a few of leucine-rich repeats (LRRs), which all are common in typical TLRs. McTLRw transcripts were constitutively expressed in all examined tissues with higher expression levels in immune related tissues, and were significantly induced in haemocytes with the challenges of live Vibrio alginolyticus, lipopolysaccharide (LPS), peptidoglycans (PGN) and ß-glucan (GLU), but not induced by polyinosinic-polycytidylic acid (poly I:C). rMcTLRw exhibited affinity to LPS, PGN and GLU while no affinity to poly I:C. Further, the downstream of TLR signaling pathway myeloid differentiation factor 88a (MyD88a), interleukin-1 receptor-associated kinase-4 (IRAK4) and tumor necrosis factor receptor-associated factor 6 (TRAF6) were significantly repressed in McTLRw silenced mussels while challenged with LPS. These results collectively indicated that McTLRw is one member of TLR family and involved in immune response to against invaders by taking participate in TLR mediated signaling pathway.


Assuntos
Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Mytilus/genética , Mytilus/imunologia , Receptores Toll-Like/genética , Receptores Toll-Like/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Perfilação da Expressão Gênica , Lipopolissacarídeos/farmacologia , Peptidoglicano/farmacologia , Filogenia , Receptores Toll-Like/química , Vibrio alginolyticus/fisiologia , beta-Glucanas/farmacologia
20.
Molecules ; 24(7)2019 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-30935016

RESUMO

Glucans are part of a group of biologically active natural molecules and are steadily gaining strong attention not only as an important food supplement, but also as an immunostimulant and potential drug. This paper represents an up-to-date review of glucans (ß-1,3-glucans) and their role in various immune reactions and the treatment of cancer. With more than 80 clinical trials evaluating their biological effects, the question is not if glucans will move from food supplement to widely accepted drug, but how soon.


Assuntos
Adjuvantes Imunológicos/química , Antineoplásicos/química , Suplementos Nutricionais , beta-Glucanas/química , Adjuvantes Imunológicos/farmacologia , Agaricales/química , Animais , Antineoplásicos/farmacologia , Regulação da Expressão Gênica , Humanos , Sistema Imunitário/efeitos dos fármacos , Valor Nutritivo , Transdução de Sinais , beta-Glucanas/farmacologia
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