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1.
Am J Chin Med ; 48(6): 1491-1509, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32924531

RESUMO

Astragalus membranaceus is the most popular traditional Chinese medicine for managing vital energy deficiency. Its injectable polysaccharide PG2 has been used for relieving cancer-related fatigue, and PG2 has immune-modulatory and anti-inflammatory effects. In this study, we explored the effects of PG2 in lung adenocarcinoma A549 and CL1-2 cells and investigated its anticancer activity, and the results were validated in severe combined immunodeficiency (SCID) mice. Although PG2 did not inhibit the growth of these cells, it dose-dependently suppressed their migration and invasion, accompanied by reduced vimentin and AXL and induced epithelial cadherin (E-cadherin) expression. Regarding the underlying molecular mechanism, PG2 treatment reduced the macrophage migration inhibitory factor (MIF), an inflammatory cytokine that promotes the epithelial-mesenchymal transition and aggressiveness of cancer cells. Consistent with the previous finding that MIF regulates matrix metalloproteinase-13 (MMP-13) and AMP-activated protein kinase (AMPK), treatment with PG2 reduced MMP-13 and activated AMPK in A549 and CL1-2 cells in this study. In SCID mice injected with A549 cells through the tail vein, intraperitoneal injection with PG2 reduced lung and abdominal metastases in parallel with decreased immunohistochemical staining of AXL, vimentin, MMP-13, and MIF in the tumor. Collectively, data revealed a potential application of PG2 in integrative cancer treatment through the suppression of MIF in cancer cells and their aggressiveness.


Assuntos
Adenocarcinoma/patologia , Astragalus propinquus/química , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Oxirredutases Intramoleculares/metabolismo , Neoplasias Pulmonares/patologia , Fatores Inibidores da Migração de Macrófagos/metabolismo , Fitoterapia , Polissacarídeos/administração & dosagem , Polissacarídeos/farmacologia , Células A549 , Adenocarcinoma/metabolismo , Animais , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Humanos , Injeções Intraperitoneais , Neoplasias Pulmonares/metabolismo , Camundongos SCID , Invasividade Neoplásica , Polissacarídeos/isolamento & purificação , Polissacarídeos/uso terapêutico
2.
Molecules ; 25(18)2020 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-32899754

RESUMO

The emergence of the Coronavirus Disease 2019 (COVID-19) caused by the SARS-CoV-2 virus has led to an unprecedented pandemic, which demands urgent development of antiviral drugs and antibodies; as well as prophylactic approaches, namely vaccines. Algae biotechnology has much to offer in this scenario given the diversity of such organisms, which are a valuable source of antiviral and anti-inflammatory compounds that can also be used to produce vaccines and antibodies. Antivirals with possible activity against SARS-CoV-2 are summarized, based on previously reported activity against Coronaviruses or other enveloped or respiratory viruses. Moreover, the potential of algae-derived anti-inflammatory compounds to treat severe cases of COVID-19 is contemplated. The scenario of producing biopharmaceuticals in recombinant algae is presented and the cases of algae-made vaccines targeting viral diseases is highlighted as valuable references for the development of anti-SARS-CoV-2 vaccines. Successful cases in the production of functional antibodies are described. Perspectives on how specific algae species and genetic engineering techniques can be applied for the production of anti-viral compounds antibodies and vaccines against SARS-CoV-2 are provided.


Assuntos
Antivirais/farmacologia , Produtos Biológicos/farmacologia , Chlamydomonas reinhardtii/genética , Infecções por Coronavirus/tratamento farmacológico , Lectinas/farmacologia , Pneumonia Viral/tratamento farmacológico , Polifenóis/farmacologia , Polissacarídeos/farmacologia , Antivirais/química , Antivirais/isolamento & purificação , Betacoronavirus/efeitos dos fármacos , Betacoronavirus/patogenicidade , Produtos Biológicos/química , Produtos Biológicos/isolamento & purificação , Núcleo Celular/química , Núcleo Celular/genética , Núcleo Celular/metabolismo , Chlamydomonas reinhardtii/química , Chlamydomonas reinhardtii/metabolismo , Cloroplastos/química , Cloroplastos/genética , Cloroplastos/metabolismo , Infecções por Coronavirus/prevenção & controle , Engenharia Genética/métodos , Humanos , Lectinas/química , Lectinas/isolamento & purificação , Coronavírus da Síndrome Respiratória do Oriente Médio/efeitos dos fármacos , Coronavírus da Síndrome Respiratória do Oriente Médio/patogenicidade , Pandemias , Polifenóis/química , Polifenóis/isolamento & purificação , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Vírus da SARS/efeitos dos fármacos , Vírus da SARS/patogenicidade , Síndrome Respiratória Aguda Grave/tratamento farmacológico , Vacinas Virais/biossíntese , Vacinas Virais/farmacologia
3.
J Chromatogr A ; 1625: 461194, 2020 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-32709309

RESUMO

Quantitative analysis of glycans released from glycoproteins using high-performance liquid chromatography (HPLC) requires fluorescent tag labeling to enhance sensitivity and selectivity. However, the methods required to remove large amounts of excess labeling reagents from the reaction mixture are time-consuming. Furthermore, these methods, including solvent extraction and solid phase extraction (SPE), often impair quantitative analysis. Here, we developed an online sample cleanup procedure for HPLC analysis of 2-aminopyridine (AP)-labeled glycans using a six-port/two-way valve and two small columns: one packed with a strong cation exchange resin (SCX) and the other comprising ODS silica gel. AP-labeled glycans delivered from an injection port were separated from excess AP by passing through an SCX column (4.6 mm i.d., 1 cm long) regulated to 40°C. The AP-labeled glycans were trapped on an ODS column (4.6 mm i.d., 1 cm long) to further separate them from inorganic contaminants. By changing the valve position after 2 min to connect the ODS column to an analysis column, AP-labeled glycans trapped in the ODS column were eluted with an acetonitrile-containing eluent followed by hydrophilic interaction liquid chromatography (HILIC) separation on an amide column or reversed-phase mode separation on a C30 column. This method was successfully used to analyze N-linked glycans released from several glycoprotein samples.


Assuntos
Aminopiridinas/química , Cromatografia Líquida de Alta Pressão/métodos , Polissacarídeos/química , Cromatografia por Troca Iônica , Cromatografia de Fase Reversa , Glicoproteínas/metabolismo , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Peptídeo-N4-(N-acetil-beta-glucosaminil) Asparagina Amidase/metabolismo , Polissacarídeos/isolamento & purificação , Extração em Fase Sólida
4.
PLoS One ; 15(6): e0232972, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32512581

RESUMO

Various dietary fibers are considered to prevent obesity by modulating the gut microbiota. Cordyceps sinensis polysaccharide (CSP) is a soluble dietary fiber known to have protective effects against obesity and related diseases, but whether these effects induce any side effects remains unknown. The function and safety of CSP were tested in high-fat diet (HFD)-feding C57BL/6J mice. The results revealed that even though CSP supplementation could prevent an increase in body weight, it aggravated liver fibrosis and steatosis as evidenced by increased inflammation, lipid metabolism markers, insulin resistance (IR) and alanine aminotransferase (ALT) in HFD-induced obesity. 16S rDNA gene sequencing was used to analyze the gut microbiota composition, and the relative abundance of the Actinobacteria phylum, including the Olsenella genus, was significantly higher in CSP-treated mice than in HFD-fed mice. CSP supplementation may increase the proportion of Actinobacteria, which can degrade CSP. The high level of Actinobacteria aggravated the disorder of the intestinal flora and contributed to the progression from obesity to nonalcoholic steatohepatitis (NASH) and related diseases.


Assuntos
Cordyceps , Dieta Hiperlipídica/efeitos adversos , Fibras na Dieta/administração & dosagem , Microbioma Gastrointestinal , Hepatopatia Gordurosa não Alcoólica/metabolismo , Polissacarídeos/administração & dosagem , Tecido Adiposo/metabolismo , Tecido Adiposo/patologia , Animais , Peso Corporal , Cordyceps/metabolismo , Modelos Animais de Doenças , Inflamação/etiologia , Inflamação/metabolismo , Resistência à Insulina , Lipídeos/sangue , Fígado/metabolismo , Fígado/patologia , Masculino , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/patologia , Polissacarídeos/isolamento & purificação
5.
Arch Biochem Biophys ; 687: 108369, 2020 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-32335047

RESUMO

A neutral tea polysaccharide (TPSN) was isolated from green tea. Gas chromatography analysis showed that TPSN was composed of d-glucose, l-arabinose and d-galactose residues at a molar ratio of 90.0: 9.1: 0.9. The weight-averaged molecular weight of TPSN was determined as about 2.0 × 105 g mol-1 using static light scattering analysis. The result of nuclear magnetic resonance (NMR) spectroscopy indicated that TPSN and water-soluble starch had similar structures. TPSN exhibited inhibitory activity towards α-amylase through the noncompetitive inhibition mechanism, but the tertiary structure of α-amylase related to enzymatic activity, analyzed using circular dichroism spectroscopy, was not affected by TPSN. Meanwhile, TPSN exhibited hydrolysis properties catalyzed by α-amylase. Molecular docking analysis revealed that the various behaviors of TPSN to α-amylase could be attributed to that the different chain segments of TPSN combined with different amino acid residues of α-amylase.


Assuntos
Inibidores Enzimáticos/química , Polissacarídeos/química , Chá/química , alfa-Amilases/antagonistas & inibidores , Animais , Camellia sinensis/química , Ensaios Enzimáticos , Inibidores Enzimáticos/isolamento & purificação , Inibidores Enzimáticos/metabolismo , Hidrólise , Cinética , Simulação de Acoplamento Molecular , Peso Molecular , Polissacarídeos/isolamento & purificação , Polissacarídeos/metabolismo , Ligação Proteica , Suínos , alfa-Amilases/metabolismo
6.
Mar Drugs ; 18(4)2020 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-32331442

RESUMO

The enzymatic depolymerization of fucoidans from brown algae allowed the production of their standardized derivatives with different biological activities. This work aimed to compare the antiviral activities of native (FeF) and modified with enzyme (FeHMP) fucoidans from F. evanescens. The cytotoxicity and antiviral activities of the FeF and FeHMP against herpes viruses (HSV-1, HSV-2), enterovirus (ECHO-1), and human immunodeficiency virus (HIV-1) in Vero and human MT-4 cell lines were examined by methylthiazolyltetrazolium bromide (MTT) and cytopathic effect (CPE) reduction assays, respectively. The efficacy of fucoidans in vivo was evaluated in the outbred mice model of vaginitis caused by HSV-2. We have shown that both FeF and FeHMP significantly inhibited virus-induced CPE in vitro and were more effective against HSV. FeF exhibited antiviral activity against HSV-2 with a selective index (SI) > 40, and FeHMP with SI ˃ 20, when they were added before virus infection or at the early stages of the HSV-2 lifecycle. Furthermore, in vivo studies showed that after intraperitoneal administration (10 mg/kg), both FeF and FeHMP protected mice from lethal intravaginal HSV-2 infection to approximately the same degree (44-56%). Thus, FeF and FeHMP have comparable potency against several DNA and RNA viruses, allowing us to consider the studied fucoidans as promising broad-spectrum antivirals.


Assuntos
Antivirais/farmacologia , Fucus/química , Polissacarídeos/farmacologia , Vírus/efeitos dos fármacos , Animais , Antivirais/isolamento & purificação , Chlorocebus aethiops , Vírus de DNA/efeitos dos fármacos , Modelos Animais de Doenças , Feminino , Humanos , Camundongos , Polissacarídeos/isolamento & purificação , Vírus de RNA/efeitos dos fármacos , Vaginite/tratamento farmacológico , Vaginite/virologia , Células Vero
7.
Food Chem ; 324: 126857, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32344342

RESUMO

Glucagon-like peptide-1 (GLP-1) is an important signal in the peripheral and neural systems, which contributes to the maintenance of glucose and energy homeostasis. In this study, 1H NMR validated polyphenols and polysaccharides extracted from sprouted quinoa yoghurt were used as isolates and conjugates to upregulate the stimulation of GLP-1 release in NCI-H716 cells. In addition, we explored their effect on proglucagon and prohormone convertase 3 mRNA expressions, HNF-3γ and CCK-2R gene protein expression, as well as cytosolic calcium release. Variations in concentration showed a dose-dependent GLP-1 stimulation, and were significantly optimized by germination. Proglucagon mRNA expression in NCI-H716 cells was upregulated, and was relatively highest with QYPSP1 treatments in a 2.68 fold. The results suggested that the conjugates had greater potential to stimulate GLP-1 release than their isolates. Sprouted quinoa yoghurt could therefore be a potential functional food useful to regulate glucose and energy homeostasis.


Assuntos
Chenopodium quinoa/química , Inibidores da Dipeptidil Peptidase IV/química , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Polifenóis/química , Polissacarídeos/química , Iogurte/análise , Cálcio/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Chenopodium quinoa/crescimento & desenvolvimento , Chenopodium quinoa/metabolismo , Cromatografia Líquida de Alta Pressão , Inibidores da Dipeptidil Peptidase IV/metabolismo , Inibidores da Dipeptidil Peptidase IV/farmacologia , Germinação , Humanos , Polifenóis/análise , Polifenóis/isolamento & purificação , Polissacarídeos/análise , Polissacarídeos/isolamento & purificação , Proglucagon/antagonistas & inibidores , Proglucagon/genética , Proglucagon/metabolismo , Pró-Proteína Convertase 1/antagonistas & inibidores , Pró-Proteína Convertase 1/genética , Pró-Proteína Convertase 1/metabolismo
8.
J Chromatogr A ; 1620: 461001, 2020 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-32151415

RESUMO

Mass spectrum (MS) is one of the most commonly used tools for qualitative and quantitative analysis of glycans. However, due to the complexity of biological samples and the low ionization efficiency of glycans, these need to be purified and derivatized prior to MS analysis. Existing purification strategies require a combination of multiple methods and are cumbersome to operate. Here, we propose a new method for the purification of glycoprotein N/O-glycans and their derivatives using a hand-packed absorbent cotton hydrophilic interaction chromatography column (HILIC). The method's reliability and applicability were verified by purifying N/O-glycans and the derivatives of standard glycoproteins, such as chicken albumin and porcine stomach mucin. Stable isotope labelling was used to compare the glycans' recovery following different purification methods. Absorbent cotton HILIC was also successfully applied for the analysis of human serum and fetal bovine serum glycoprotein N-glycans. Finally, testing revealed high binding capacity (9 mg/g-1 maltohexaose/absorbent cotton) and good recovery (average recovery was 91.7%) of glycans. Compared with traditional procedures, the proposed purification method offers considerable advantages, such as simplicity, high efficiency, economy, universality, and broad applicability for the pretreatment of glycans and their derivatives in biological samples prior to MS analysis.


Assuntos
Cromatografia/métodos , Polissacarídeos/isolamento & purificação , Animais , Galinhas , Fibra de Algodão , Glicoproteínas/química , Humanos , Interações Hidrofóbicas e Hidrofílicas , Marcação por Isótopo , Espectrometria de Massas , Mucinas/química , Polissacarídeos/sangue , Polissacarídeos/química , Suínos
9.
Food Chem ; 316: 126371, 2020 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-32062579

RESUMO

Two novel selenium polysaccharide fractions (SeTPS-1 and SeTPS-2) were isolated purified, characterized from Se-enriched tea. The results showed that the molecular weights and Se content of SeTPS-1 and SeTPS-2 were 1.7 × 104 Da, 1.3 × 104 Da, and 23.50 µg/g and 13.47 µg/g, respectively. SeTPS-1 and SeTPS-2 had absorption spectra typical of selenium esters. SeTPS-1 was composed of glucose and galactose at a molar ratio of 80.1:2.3, respectively, while SeTPS-2 was composed of arabinose, glucose, galactose and galacturonic acid with a molar ratio of 2.04: 48.83: 3.21: 1.30, respectively. Both SeTPS-1 and SeTPS-2 adopted a random coil conformation. Importantly, in vitro assessment of the antioxidant capacity revealed that SeTPS-1 is a more potent antioxidant compared to SeTPS-2. Both compounds were effective at reducing DNA damage induced by H2O2. The promising data suggesting that these compounds confer natural protection against DNA-damaging agents, thereby contributing to the functional food qualities of tea.


Assuntos
Polissacarídeos/isolamento & purificação , Selênio/análise , Chá/química , Antioxidantes/farmacologia , Peso Molecular , Polissacarídeos/química
10.
Carbohydr Polym ; 234: 115895, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32070515

RESUMO

Fucosylated chondroitin sulfate (FCS) HeSp was isolated from the Patagonian sea cucumber Hemioedema spectabilis. Ion-exchange chromatography was applied for purification of the FCS from the crude extract of sulfated polysaccharides. Analysis of monosaccharide and sulfate content of HeSp revealed the molar ratio of GlcA:GalNAc:Fuc:SO3Na as 1.15:1:1.1:3.9. Molecular weight of HeSp (44.1 kDa) was determined by GPC. According to the NMR spectral data, the main fragment of HeSp was the trisaccharide →3)-ß-d-GalNAc-(1→4)-ß-d-GlcA(3-O-α-l-Fuc)-(1→, where GalNAc units were sulfated either at O-4, at O-6 or both at O-4 and O-6. The fucosyl branches attached to O-3 of GlcA showed also different patterns of sulfation: Fucp2S4S, Fucp4S and Fucp3S4S were found in a ratio of 3.8:1.5:1. Besides, small amounts of the disaccharide fragment →3)-ß-d-GalNAc-(1→4)-ß-d-GlcA3S-(1→ were observed in a structure of HeSp. The polysaccharide was found to block cancer cells adhesion to platelet-coated surface and to inhibit tubulogenesis, thus demonstrating the potential antitumor activity.


Assuntos
Antineoplásicos/farmacologia , Sulfatos de Condroitina/farmacologia , Polissacarídeos/farmacologia , Pepinos-do-Mar/química , Animais , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Adesão Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sulfatos de Condroitina/química , Sulfatos de Condroitina/isolamento & purificação , Ensaios de Seleção de Medicamentos Antitumorais , Células HeLa , Humanos , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Especificidade da Espécie , Células Tumorais Cultivadas
11.
Carbohydr Polym ; 234: 115896, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32070516

RESUMO

The preparation mainly composed of extraction, pre-purification and dehydration is essential for the research and development of natural polysaccharides. The methods or conditions used in the three procedures had significant effects on the composition, structure and function of the polysaccharides obtained. Temperature, pH, enzyme, ultrasound and microwave were the important factors associated with their physicochemical changes. Molecular degradation and intermolecular interaction were two of the main mechanisms responsible for the changes. The degradations of polysaccharides responding to hydrothermal and ultrasonic conditions could be partly descripted by multiple linear regression model, implying the possibility for the prediction and control of polysaccharide degradation. Moreover, the interactions between polysaccharide and other compounds, forming complexes natively or conditionally, could be selectively triggered or eliminated to obtain polysaccharides under certain functions. This work shows new insights into the preparation of polysaccharides, which could benefit the efficient utilization of their natural and modified properties.


Assuntos
Produtos Biológicos/química , Produtos Biológicos/metabolismo , Polissacarídeos/química , Polissacarídeos/metabolismo , Produtos Biológicos/isolamento & purificação , Físico-Química , Modelos Lineares , Polissacarídeos/isolamento & purificação
12.
Carbohydr Polym ; 234: 115906, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32070525

RESUMO

The present study was to investigate the molecular mechanism underlying lymphocyte activation by total polysaccharides from Atractylodis macrocephalae (RAMPtp). The results showed that RAMPtp significantly promoted the secretions of cytokines (IFN-γ, IL-1α, IL-21, IFN-α, CCL4, CXCL9 and CXCL10), increased the proportions of CD4+ and CD8+ subpopulations, and enhanced the expressions of c-JUN, NFAT4, STAT1 and STAT3. microRNA sequencing identified 67 differentially expressed miRNAs (DEMs) in RAMPtp-stimulated SMLN lymphocytes, including 55 up-regulated and 12 down-regulated. GO and KEGG enrichment analyses of the predicted DEMs-targeted genes indicated that they were associated with immune system pathways, including PI3K-Akt, MAPKs, Jak-STAT and Calcium signaling pathways, which were confirmed by western blot and pathway inhibition assays. RAMPtp was further observed to favor immunostimulatory effect on both T and B lymphocytes via binding to TCR and membrane Ig individually. These findings might explain the immunomodulatory mechanism of RAMPtp in ameliorating the bovine intramammary infection.


Assuntos
Atractylodes/química , Linfócitos B/efeitos dos fármacos , Polissacarídeos/farmacologia , Linfócitos T/efeitos dos fármacos , Receptor 4 Toll-Like/metabolismo , Animais , Bovinos , Biologia Computacional , Citocinas/biossíntese , Ativação Linfocitária/efeitos dos fármacos , Tamanho da Partícula , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Transdução de Sinais/efeitos dos fármacos , Propriedades de Superfície
13.
Food Chem ; 315: 126266, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32000083

RESUMO

Flaxseeds are widely consumed for their desirable sensory attributes and health benefits. We focused on enhancing the sustainability and economic potential of flaxseeds by characterizing functional attributes of polysaccharides isolated from flaxseed hull residues. In particular, antioxidant and immune-stimulatory polysaccharides were isolated and purified from flaxseed hull. Infrared spectroscopy was used to identify the key functional groups. The polysaccharides were composed of mannose, rhamnose, galactose, glucose, galactose, xylose, arabinose, and fucose. In vitro studies showed certain flaxseed hull polysaccharide fractions exhibited strong antioxidant activities, increased nitric oxide levels, and enhanced the production of cytokines (TNF-α and IL-6). In the presence of 200 µg/mL of one of these fractions, the levels of p-ERK, p-JNK, and p-p38 increased significantly by 1.8-, 9.0-, and 6.7-fold. These polysaccharide fractions may exhibit their immune-regulatory properties partly by modulating the MAPK pathway. The flaxseed hull polysaccharides identified have potential application as natural antioxidants and immune-enhancing nutraceuticals.


Assuntos
Adjuvantes Imunológicos/farmacologia , Antioxidantes/farmacologia , Linho/química , Polissacarídeos/análise , Polissacarídeos/farmacologia , Adjuvantes Imunológicos/química , Animais , Antioxidantes/análise , Antioxidantes/química , Antioxidantes/isolamento & purificação , Citocinas/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Camundongos , Óxido Nítrico/biossíntese , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Células RAW 264.7 , Relação Estrutura-Atividade
14.
Food Chem ; 313: 126164, 2020 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-31935662

RESUMO

A thermoseparating aqueous two-phase system (ATPS) based on ethylene oxide-propylene oxide (EOPO) copolymer and deep eutectic solvents (DES) was developed for the extraction and preliminary purification of polysaccharides in Camellia oleifera Abel. seed cake. DESs were used as the extraction media for obtaining the crude extract. The maximum extraction yield (Y) of polysaccharides was 152.37 mg/g in the crude extract. Then the crude polysaccharides were extracted and preliminary purified by EOPO/DES ATPS. The maximum extraction efficiency (E.E.) of polysaccharides was 86.91% in EOPO-rich phase by the first ATPS extraction. The EOPO-rich phase was separated and the polysaccharides were recovered by temperature-induced phase separation. The maximum recovery efficiency (R.E.) was 84.92% in aqueous phase by the second ATPS extraction. This thermoseparating ATPS possesses the merits of good environment-friendlies, simple operation, and easy recovery of components, which can be used as an efficient method for the extraction and separation of polysaccharides.


Assuntos
Camellia/química , Fracionamento Químico/métodos , Polissacarídeos/isolamento & purificação , Solventes/química , Manipulação de Alimentos/métodos , Polietilenos/química , Polipropilenos/química , Polissacarídeos/química , Sementes/química , Temperatura , Água
15.
Carbohydr Polym ; 231: 115688, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31888812

RESUMO

Two new isolated polysaccharides, ARPP-40 (40 % ethanol precipitate) and ARPP-70 (70 % ethanol precipitate), were extracted from Anoectochilus roxburghii. The physicochemical properties of two polysaccharides were analyzed and the results showed the relative weight average molecular weights and contents of neutral sugar for ARPP-40 and ARPP-70 were 423 kDa and 97.4 %, 10.8 kDa and 51.4 %, respectively. In terms of monosaccharide composition, ARPP-40 contained only glucose, while ARPP-70 was composed of seven monosaccharides, of which glucose and galactose were the main components. Furthermore, the structure and conformation characteristics of ARPP-40 were systematically investigated. The results revealed that ARPP-40 was supposed to be a glucan and existed as a flexible chain with a polydispersity index of 1.02 in 0.1 M NaNO3 solution. The systematic information on structural and conformational properties of ARPP-40 was meaningful for its further application in food and medicinal industry.


Assuntos
Orchidaceae/química , Polissacarídeos/química , Açúcares/química , Antioxidantes/química , Fracionamento Químico , Galactose/química , Glucose/química , Peso Molecular , Monossacarídeos/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/ultraestrutura , Açúcares/isolamento & purificação
16.
Carbohydr Polym ; 231: 115732, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31888819

RESUMO

Cibotium barometz, an important traditional Chinese medicine, is used in strengthening bones and tendons. We found that C. barometz crude polysaccharides (CB70) could alleviate bone loss and markedly improve the biomechanical properties of OVX rats. Thus, to clarify biological active ingredient(s) of CB70, two homogeneous polysaccharides (CBP70-1-1 and CBP70-1-2) were purified from CB70. A combination of monosaccharide composition, FT-IR, GC-MS and NMR analysis indicated that CBP70-1-1 was composed of →6)-D-Galp-(1→, D-Glcp-(1→, →3,6)-D-Manp-(1→, →4)-D-Glcp-(1→ and →6)-D-Glcp-(1→ with relative molecular weights of 12,724 Da, and CBP70-1-2 was composed of →4)-D-Glcp-(1→, D-Glcp-(1→, →3,6)-D-Manp-(1→, →6)-D-Galp-(1→, →4,6)-D-Glcp-(1→ and →3)-L-Araf-(1→ with relative molecular weights of 3611 Da. Morphological analyses revealed that CBP70-1-1 and CBP70-1-2 appeared as a sheet that were irregular in size and shape, while the surface of CBP70-1-1 was full of sharp protuberances and CBP70-1-2 was smooth. Furthermore, the effects of CBP70-1-1 and CBP70-1-2 on the proliferation, differentiation and mineralization of mouse pre-osteoblastic MC3T3-E1 cells were assessed via CCK-8 assay, alkaline phosphatase activity assay, and alizarin red-based assay, respectively. These results revealed that CBP70-1-1 and CBP70-1-2 significantly promoted the proliferation, differentiation and mineralization of MC3T3-E1 cells, even better than E2. More importantly, quantitative real-time PCR and Western blot analysis indicated that CBP70-1-2 pronouncedly promoted the expression of osteogenic-related marker genes (Runx2, Osx, Ocn and Opn) and proteins (BMP2, RUNX2, OSX and p-SMAD1), which implies that the osteogenic activity of CBP70-1-2 is accomplished mainly by activating the BMP2/SMAD1 signaling pathway. These findings suggest CBP70-1-2 as a potential natural anti-osteoporotic agent for pharmacotherapy.


Assuntos
Embriófitas/química , Osteogênese/efeitos dos fármacos , Osteoporose/tratamento farmacológico , Polissacarídeos/química , Células 3T3 , Animais , Proteína Morfogenética Óssea 2/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Camundongos , Osteoporose/patologia , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Rizoma/química , Transdução de Sinais/efeitos dos fármacos , Proteína Smad1/genética , Espectroscopia de Infravermelho com Transformada de Fourier
17.
Carbohydr Polym ; 231: 115728, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31888838

RESUMO

A water-soluble acidic polysaccharide, XB-PS3, was isolated from the twigs of Juniperus pingii var. Wilsonii with a molecular weight of 86.04 kDa. By means of monosaccharide composition analysis, methylation, 2D NMR spectroscopy and UPLC-MS analysis, we concluded that XB-PS3 had a backbone composed of →2,4)-α-Manp-(1→ and →4)-α-GalpA-(1→ (60 % esterified), with an araban branch attached to O-2 of →2,4)-α-Manp-(1→. The possible repeating units were further validated by oligosaccharide analysis and partial acid hydrolysis. XB-PS3 exhibited potent anticomplement activity with CH50 value of 117.23 ± 18.74 µg/mL and interacted with C3, C4, C5 and C9 in the complement activation cascade. However, the anticomplement activity was significantly weakened when the galacturonic acids were reduced (CH50: 268.55 ± 16.82 µg/mL) or the branches were removed by partial hydrolysis (CH50: 197.76 ± 21.81 µg/mL), indicating the important role of uronic acids and branch structure in the polysaccharide's anticomplement activity.


Assuntos
Proteínas Inativadoras do Complemento/química , Juniperus/química , Polissacarídeos/química , Ácidos/química , Cromatografia Líquida , Complemento C3/antagonistas & inibidores , Complemento C3/química , Proteínas Inativadoras do Complemento/isolamento & purificação , Proteínas Inativadoras do Complemento/farmacologia , Carboidratos da Dieta/farmacologia , Humanos , Hidrólise , Peso Molecular , Monossacarídeos/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Espectrometria de Massas em Tandem , Ácidos Urônicos/química , Água/química
18.
Carbohydr Polym ; 232: 115766, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-31952583

RESUMO

Type 2 diabetes mellitus plagues many people in China and the world, and its nephritis complication is the leading cause of death for patients. Paecilomyces hepiali contained various functional components, especially polysaccharides, which possesses well pharmacological activities. In this study, polysaccharide purified from Paecilomyces hepiali fermented mycelium entitled PHEA was obtained, and its structure was systemically characterized using fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance (NMR). In C57BL/KsJ (BKS).Cg-Dock7m +/+ Leprdb/JNju mice (db/db mice), via detecting the alternations on biochemical criterions, pathological indicators and protein expressions related to nuclear factor-E2-related factor 2 (Nrf2) and nuclear factor-κB (NF-κB) signaling in serum and/or tissues including muscle, liver and kidney after 8-week PHEA administration, the hypoglycemic, hypolipidemic, and anti-diabetic nephropathic activities of PHEA were confirmed. The purified P. Hepiali polysaccharide with the anti-diabetic and -nephritic properties was first reported in this study via regulating Nrf2-meadited NF-κB signaling in db/db mice.


Assuntos
Diabetes Mellitus Tipo 2/tratamento farmacológico , Nefropatias Diabéticas/tratamento farmacológico , Hipoglicemiantes/uso terapêutico , Micélio/química , Polissacarídeos/uso terapêutico , Animais , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Nefropatias Diabéticas/metabolismo , Modelos Animais de Doenças , Fermentação , Hipoglicemiantes/química , Hipoglicemiantes/isolamento & purificação , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Micélio/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Tamanho da Partícula , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Transdução de Sinais/efeitos dos fármacos , Propriedades de Superfície
19.
Carbohydr Polym ; 230: 115680, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31887951

RESUMO

Three main fractions, one neutral polysaccharide (ACPN-1a) and two acidic polysaccharides (ACPA-1a and ACPA-2a), were isolated from crude polysaccharides of Auricularia cornea var. Li. (ACP). The physicochemical properties and structural characteristics of these fractions were investigated using High Performance Gel Permeation Chromatography (HPGPC), High Performance Liquid Chromatography (HPLC), Scanning Electron Microscopy (SEM), Fourier Transform Infrared (FTIR), and Nuclear Magnetic Resonance (NMR) spectra. The results showed that ACPN-1a with a higher molecular weight (∼2.18 × 106 Da) was a ß-(1→3)-glucan with (1→6)-linked ß-Glcp residues for side chains. ACPA-1a was a heteropolysaccharide and had the backbone of (1→3)-linked ß-Glcp and (1→6)-linked α-Manp and side chains with ß-Xylp or ß-GlcpA residues. ACPA-2a had the average molecular weight of 8.5 × 105 Da and was composed of a (1→6)-linked α-Manp backbone and side chains with ß-Xylp or ß-GlcpA residues. The data provided useful information for further investigation on the structure-activity relationship of Auricularia cornea var. Li. polysaccharides.


Assuntos
Basidiomycota/metabolismo , Polissacarídeos/química , Polissacarídeos/isolamento & purificação
20.
Carbohydr Polym ; 230: 115561, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31887876

RESUMO

The current hydrocolloid industry requires new techniques for biomass characterization, which can quickly and ecologically characterize contained sugars. This work proposes the use of Fourier Transform Infrared microspectroscopy in combination with multivariate methods, to localize and identify the main carbohydrates and other components present in fresh brown seaweeds, avoiding time-consuming samples pre-treatments. Infrared images of Macrocystis pyrifera samples were analyzed by Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) and Principal Component Analysis (PCA) as chemometrics techniques to identify the compounds. MCR-ALS was the best strategy, delivering pure spectra of chemical compound that PCA did not. The carbohydrates identified by this method were 1-3-ß-glucans divided into endofibers and laminarin; two types of fucoidans (rich in fucose or mannuronic acid), alginate and mannitol, besides other compounds such as proteins. This technique represents an opportunity for the hydrocolloid industry for a modern, rapid and environmentally-friendly characterization of macroalgal biomass to enhance its use.


Assuntos
Carboidratos/isolamento & purificação , Polissacarídeos/isolamento & purificação , Alga Marinha/química , Espectroscopia de Infravermelho com Transformada de Fourier , Alginatos/química , Carboidratos/química , Análise dos Mínimos Quadrados , Análise Multivariada , Polissacarídeos/química , Polissacarídeos/classificação , Análise de Componente Principal , Açúcares/química , Açúcares/isolamento & purificação
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