RESUMO
The aim of this study was to investigate the surface morphological features and in vivo immunomodulatory activities of a hetero polysaccharide fraction (HEP-W) from Hericium erinaceus. SEM and AFM images revealed that HEP-W displayed a flexible random coil conformation, and these flexible winding chains further formed continuous fiber network structure. Meanwhile, Congo red assay and XRD further proved that HEP-W mainly exhibited amorphous structure with non-triple-helical conformation in solution. In vivo immunomodulatory experiments demonstrated that HEP-W possessed protective effects against cyclophosphamide-induced immunosuppression in mice by significantly enhancing immune organ index, splenocyte proliferation, NK cell activity, IL-2 production as well as improving the macrophage phagocytosis. These findings suggest that HEP-W could be explored as a natural and effective immunomodulatory agent.
Assuntos
Ciclofosfamida/efeitos adversos , Ciclofosfamida/antagonistas & inibidores , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/farmacologia , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Imunossupressores/efeitos adversos , Imunossupressores/antagonistas & inibidores , Animais , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/imunologia , Configuração de Carboidratos , Vermelho Congo , Feminino , Polissacarídeos Fúngicos/ultraestrutura , Hericium/química , Interleucina-2/sangue , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Leucócitos/efeitos dos fármacos , Leucócitos/imunologia , Ativação Linfocitária/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Fagocitose/efeitos dos fármacos , Propriedades de Superfície , Difração de Raios XRESUMO
The immunomodulatory effects of the four extracellular polysaccharides, namely WPA, WPB, AP2A, and TP1A, which were isolated from the fermented broth of Aspergillus aculeatus, A. terreus and Trichoderma sp. KK19L1, were investigated in vitro. WPA, WPB, AP2A, and TP1A were not toxic to RAW264.7 cells. These polysaccharides enhanced cell viability. WPA, WPB, AP2A, and TP1A showed increased immunomodulatory effect by strengthening the phagocytic activity and enhancing the release of NO, TNF-α and IL-6 from RAW264.7 cells. WPA, WPB, AP2A, and TP1A exhibited different immunomodulatory activity in vitro due to their different structural characterizations, and their immunoregulatory effects decreased successively in the following order: WPA, WPB, AP2A, and TP1A. The extracellular polysaccharides WPA, WPB, AP2A, and TP1A had potent immunomodulatory effects and could be used as potential immunomodulatory agents in the fields of functional food and medicine.
Assuntos
Aspergillus/ultraestrutura , Polissacarídeos Fúngicos/ultraestrutura , Imunomodulação/efeitos dos fármacos , Animais , Aspergillus/química , Sobrevivência Celular/efeitos dos fármacos , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/farmacologia , Macrófagos/química , Macrófagos/efeitos dos fármacos , Macrófagos/ultraestrutura , Camundongos , Células RAW 264.7 , Fator de Necrose Tumoral alfa/genéticaRESUMO
The structural properties and the inhibition on α-d-glucosidase activity of the low molecular weight (LCMPs-II) obtained from the optimized acid hydrolysis of the Cordyceps militaris polysaccharides (CMPs) were investigated in this paper. The LCMPs-II with a molecular weight of 28 KDa mainly composed of rhamnose, xylose and glucose with the molar ratio of 1: 2.19: 6.73 was separated from LCMPs-I which was the acid hydrolysis product of CMPs by chromatography on Sephadex G-100 column. The solubility of LCMPs-II was tested to be 32.12±1.05g in 100mL distilled water under 25°C. Its solubility was almost as twice as that of CMPs. Afterward, the structural features of LCMPs-II was investigated by a combination of chemical and instrumental analysis such as the specific rotation determination, FT-IR, periodate oxidation-Smith degradation, Congo-red, GC, scanning electron microscope and NMR. The results showed that the optical rotation of LCMPs-II was +25° and it was 1,3-branched-rhamnoxyloglucan which had a linear backbone of (1â4)-linked α-d-glucopyranose (α-d-Glcp units).
Assuntos
Polissacarídeos Fúngicos/química , Inibidores de Glicosídeo Hidrolases/química , alfa-Glucosidases/química , Cordyceps/química , Polissacarídeos Fúngicos/ultraestrutura , Hidrólise , Peso Molecular , SolubilidadeRESUMO
A novel polysaccharide (CMPA90-1; compound 1) was isolated from the cultured fruiting bodies of Cordyceps militaris. The chemical structure of compound 1 was elucidated by acid hydrolysis, periodate oxidation, Smith degradation, and methylation analysis, along with Fourier transform infrared spectroscopy, high-performance anion-exchange chromatography coupled with pulsed amperometric detection, gas chromatography-mass spectrometry, and one-dimensional [(1)H and (13)C nuclear magnetic resonance (NMR)] and two-dimensional NMR (heteronuclear single-quantum coherence and heteronuclear multiple-bond correlation). Sulfation of compound 1 by the chlorosulfonic acid-pyridine (CSA-Pyr) method led to synthesis of its sulfated analogue (CMPA90-M1; compound 2). The ultrastructures of both compounds 1 and 2 were further characterized by scanning electron microscopy and atomic force microscopy. The results of antioxidant assays showed that compounds 1 and 2 exhibited free-radical-scavenging effects, ferrous-ion-chelating ability, and reducing power. Also, in the cytotoxicity assay, compounds 1 and 2 showed inhibitory activity against A549 cells, with IC50 values of 39.08 and 17.33 µg/mL, respectively.
Assuntos
Cordyceps/química , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/farmacologia , Sulfatos/química , Sulfatos/farmacologia , Antineoplásicos , Antioxidantes , Linhagem Celular Tumoral , Sequestradores de Radicais Livres , Carpóforos/química , Polissacarídeos Fúngicos/ultraestrutura , Humanos , Quelantes de Ferro/química , Microscopia Eletrônica de VarreduraRESUMO
AIM: Confocal laser-scanning microscopy (CLSM) was carried out to investigate the exopolysaccharide matrix of Candida albicans (C. albicans) biofilms developed on denture material under dietary carbohydrate exposure. METHODS: Biofilms were developed on poly(methyl methacrylate) discs in culture media without (control) or with supplementation by glucose or sucrose for 72 h. For the CLSM analysis, biofilms were labeled with concanavalin A (ConA) during its development. Afterwards, biofilms were also labeled with SYTO-9. To confirm the results, the matrix was investigated by the phenol-sulfuric method. Data were analyzed by anova, followed by Tukey's test, with the level of significance set at 5%. RESULTS: The use of ConA during biofilm development provided effective labeling of the exopolysaccharide matrix. The exposure to sucrose resulted in biofilms with the highest exopolysaccharide matrix biovolume (P < 0.05). The characterization obtained by CLSM was confirmed by the phenol-sulfuric method. CONCLUSION: Confocal laser-scanning microscopy was found to be an effective tool for investigating the exopolysaccharide matrix of C. albicans biofilms, and exposure to sucrose resulted in increased matrix production.