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1.
Molecules ; 27(15)2022 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-35956965

RESUMO

Porous carbohydrate materials such as tobacco shreds readily absorb moisture and become damp during processing, storage, and consumption (smoking). Traditional humectants have the ability of moisture retention but moisture-proofing is poor. Polygonatum cyrtonema Hua polysaccharide (PCP 85-1-1) was separated by fractional precipitation and was purified by anion exchange and gel permeation chromatography. The average molecular weight (Mw) of PCP 85-1-1 was 2.88 × 103 Da. The monosaccharide composition implied that PCP 85-1-1 consisted of fucose, glucose, and fructose, and the molar ratio was 22.73:33.63:43.65. When 2% PCP 85-1-1 was added to tobacco shreds, the ability of moisture retention and moisture-proofing were significantly enhanced. The moisture retention index (MRI) and moisture-proofing index (MPI) increased from 1.95 and 1.67 to 2.11 and 2.14, respectively. Additionally, the effects of PCP 85-1-1 on the aroma and taste of tobacco shreds were evaluated by electronic tongue and gas chromatography-mass spectrometry (GC-MS). These results indicated that PCP 85-1-1 had the characteristics of preventing water absorption under high relative humidity and moisturizing under dry conditions. The problem that traditional humectants are poorly moisture-proof was solved. PCP 85-1-1 can be utilized as a natural humectant on porous carbohydrates, which provides a reference for its development and utilization.


Assuntos
Polygonatum , Carboidratos da Dieta , Higroscópicos , Polygonatum/química , Polissacarídeos/química , Porosidade
2.
Appl Environ Microbiol ; 87(13): e0053321, 2021 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-33893114

RESUMO

To systemically understand the biosynthetic pathways of bioactive substances, including triterpenoids and polysaccharides, in Ganoderma lucidum, the correlation between substrate degradation and carbohydrate and triterpenoid metabolism during growth was analyzed by combining changes in metabolite content and changes in related enzyme expression in G. lucidum over 5 growth phases. Changes in low-polarity triterpenoid content were correlated with changes in glucose and mannitol contents in fruiting bodies. Additionally, changes in medium-polarity triterpenoid content were correlated with changes in the lignocellulose content of the substrate and with the glucose, trehalose, and mannitol contents of fruiting bodies. Weighted gene coexpression network analysis (WGCNA) indicated that changes in trehalose and polyol contents were related to carbohydrate catabolism and polysaccharide synthesis. Changes in triterpenoid content were related to expression of the carbohydrate catabolic enzymes laccase, cellulase, hemicellulase, and polysaccharide synthase and to the expression of several cytochrome P450 monooxygenases (CYPs). It was concluded that the products of cellulose and hemicellulose degradation participate in polyol, trehalose, and polysaccharide synthesis during initial fruiting body formation. These carbohydrates accumulate in the early phase of fruiting body formation and are utilized when the fruiting bodies mature and a large number of spores are ejected. An increase in carbohydrate metabolism provides additional precursors for the synthesis of triterpenoids. IMPORTANCE Most studies of G. lucidum have focused on its medicinal function and on the mechanism of its activity, whereas the physiological metabolism and synthesis of bioactive substances during the growth of this species have been less studied. Therefore, theoretical guidance for cultivation methods to increase the production of bioactive compounds remains lacking. This study integrated changes in the lignocellulose, carbohydrate, and triterpenoid contents of G. lucidum with enzyme expression from transcriptomics data using WGCNA. The findings helped us better understand the connections between substrate utilization and the synthesis of polysaccharides and triterpenoids during the cultivation cycle of G. lucidum. The results of WGCNA suggest that the synthesis of triterpenoids can be enhanced not only through regulating the expression of enzymes in the triterpenoid pathway, but also through regulating carbohydrate metabolism and substrate degradation. This study provides a potential approach and identifies enzymes that can be targeted to regulate lignocellulose degradation and accelerate the accumulation of bioactive substances by regulating substrate degradation in G. lucidum.


Assuntos
Metabolismo dos Carboidratos , Lignina/metabolismo , Reishi , Triterpenos/metabolismo , Carpóforos/metabolismo , Regulação Fúngica da Expressão Gênica , Reishi/genética , Reishi/crescimento & desenvolvimento , Reishi/metabolismo , Transcriptoma
3.
Adv Exp Med Biol ; 1181: 107-134, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31677141

RESUMO

Ganoderma, named lingzhi in China, has been used for centuries as drug and nutraceutical to treat diseases. Based on our research and other literatures, the chapter summarizes the progress of preparation, structural features and properties, bioactivities of Ganoderma polysaccharides. The aim is to provide a comprehensive source of information for researchers and consumers of Ganoderma, so they can better understand Ganoderma polysaccharides and their biological activities. In addition, more clinical studies should be carried out to meet the criteria for new drug development, and more convincing scientific data should be provided. In addition, on the basis of a large number of studies on Ganoderma polysaccharides, we suggest that more clinical studies should be carried out so that Ganoderma can be better recognized and applied all over the world.


Assuntos
Ganoderma , Polissacarídeos , China , Ganoderma/química , Compostos Fitoquímicos/metabolismo , Polissacarídeos/química , Polissacarídeos/metabolismo
4.
Biomed Chromatogr ; 32(10): e4283, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29748985

RESUMO

Ganoderic acid S, ganoderic acid T and ganoderal B are the main bioactive triterpenes of Ganoderma lucidum. In this study, mycelia of G. lucidum were obtained by two-stage fermentation and then extracted by ethanol and petroleum ether sequentially to obtain crude triterpenes. The crude sample was further purified by recycling high-speed counter-current chromatography with n-hexane-ethyl acetate-methanol-water (7:12:11:5, v/v/v/v) as the optimized two-phase solvent system. A 16.4 mg aliquot of ganoderol B with a purity of 90.4% was separated from 300 mg of the crude sample in a single run. After employing the recycling elution mode of HSCCC with n-hexane-ethyl acetate-methanol-water (6:10:8:4.5, v/v/v/v) for five cycles, 25.7 mg ganoderic acid T and 3.7 mg ganoderic acid S with purities of 97.8 and 83.0%, respectively, were obtained. The purities of three compounds were determined by high-performance liquid chromatography and their chemical structures were identified by NMR and MS data.


Assuntos
Ganoderma/química , Lanosterol/análogos & derivados , Triterpenos , Cromatografia Líquida de Alta Pressão , Distribuição Contracorrente , Lanosterol/análise , Lanosterol/química , Lanosterol/isolamento & purificação , Triterpenos/análise , Triterpenos/química , Triterpenos/isolamento & purificação
5.
Phytochem Anal ; 28(2): 115-124, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28028844

RESUMO

INTRODUCTION: TLC bioautography for tyrosinase inhibitors has made recent progress; however, an assay with a relative low consumption of enzyme and quantitative capability would greatly advance the efficacy of related TLC bioautographic assays. OBJECTIVE: An improved TLC bioautographic assay for detecting tyrosinase inhibitors was developed and validated in this study. METHODS: L-DOPA (better water-solubility than L-tyrosine) was used as the substrate instead of reported L-tyrosine. The effects of enzyme and substrate concentrations, reaction temperatures and times, and pH values of the reaction system as well as different plate types on the TLC bioautographic assay were optimised. The quantitative analysis was conducted by densitometric scanning of spot areas, and expressed as the relative tyrosinase inhibitory capacity (RTIC) using a positive control (kojic acid) equivalent. RESULTS: The limit of detection (LOD) of this assay was 1.0 ng for kojic acid. This assay has acceptable accuracy (101.73-102.90%), intra- and inter-day, and intra- and inter-plate precisions [relative standard deviation (RSD), less than 7.0%], and ruggedness (RSD, less than 3.5%). The consumption of enzyme (75 U/mL) is relatively low. Two tyrosinase inhibitory compounds including naringenin and 1-O-ß-D-glucopyranosyl-4-allylbenzene have been isolated from Rhodiola sacra guided by this TLC bioautographic assay. CONCLUSION: Our improved assay is a relatively low-cost, sensitive, and quantitative method compared to the reported TLC bioautographic assays. Copyright © 2016 John Wiley & Sons, Ltd.


Assuntos
Produtos Biológicos/química , Cromatografia em Camada Fina/métodos , Inibidores Enzimáticos/análise , Monofenol Mono-Oxigenase/antagonistas & inibidores , Animais , Linhagem Celular Tumoral , Camundongos , Reprodutibilidade dos Testes
6.
Phytochem Anal ; 27(1): 5-12, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26219532

RESUMO

INTRODUCTION: Lipase inhibitory assays based on TLC bioautography have made recent progress; however, an assay with greater substrate specificity and quantitative capabilities would advance the efficacy of this particular bioassay. OBJECTIVE: To address these limitations, a new TLC bioautographic assay for detecting lipase inhibitors was developed and validated in this study. METHODS: The new TLC bioautographic assay was based on reaction of lipase with ß-naphthyl myristate and the subsequent formation of the purple dye between ß-naphthol and Fast Blue B salt (FBB). The relative lipase inhibitory capacity (RLIC) was determined by a TLC densitometry with fluorescence detection, expressed as orlistat equivalents in millimoles on a per sample weight basis. Six pure compounds and three natural extracts were evaluated for their potential lipase inhibitory activities by this TLC bioautographic assay. RESULTS: The ß-naphthyl myristate as the substrate improved the detection sensitivity and specificity significantly. The limit of detection (LOD) of this assay was 0.01 ng for orlistat, the current treatment for obesity. This assay has acceptable accuracy (92.07-105.39%), intra-day and inter-day precisions [relative standard deviation (RSD), 2.64-4.40%], as well as intra-plate and inter-plate precisions (RSD, 1.8-4.9%). CONCLUSION: The developed method is rapid, simple, stable, and specific for screening and estimation of the potential lipase inhibitors.


Assuntos
Cromatografia em Camada Fina/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , Inibidores Enzimáticos/análise , Lipase/antagonistas & inibidores , Cromatografia em Camada Fina/instrumentação , Compostos de Diazônio/análise , Compostos de Diazônio/química , Inibidores Enzimáticos/farmacologia , Desenho de Equipamento , Hidrólise , Lactonas/análise , Limite de Detecção , Lipase/metabolismo , Miristatos/química , Miristatos/metabolismo , Orlistate , Extratos Vegetais/análise , Extratos Vegetais/farmacologia , Reprodutibilidade dos Testes
7.
Guang Pu Xue Yu Guang Pu Fen Xi ; 35(5): 1331-4, 2015 May.
Artigo em Zh | MEDLINE | ID: mdl-26415455

RESUMO

Chaenomeles speciosa fruits were extracted using water. The extracts were precipitated with 20%~95% (φ) ethanol, respectively. The amount of total polysaccharide was measured with phenol-sulfuric acid method. A method using high-performance size-exclusion chromatography (HPSEC) equipped with multiangle laser-light-scattering photometry (MALLS) and differential refractometry (RI) was presented for determining the molecular weight and molecular weigh distribution. RAW264.7 macrophage were cultured and stimulated with the polysaccharides in vitro and the production of nitric oxide in the cells was determined by the Griess assay. The aim of the study is to determine the amount and the molecular weight of the polysaccharides from Chaenomeles speciosa fruits, and preliminary investigate the immunomodulatory activity, The study provided the basis datas for the further research of Chaenomeles speciosa fruits. , and provided a simple and system method for the research of natural polysaccharide. The ethanol fractional precipitation showed that the order of total polysaccharide content was 95%>80%>40% ≥60%>20%. The results indicated that most polysaccharide from Chaenomeles speciosa fruits might be precipitated when ethanol concentration was up to 95% (T) and the crude polysaccharide purity had risen from 35. 1% to 45. 0% when the concentration of ethanol increased from 20% to 95%. HPSEC-MALLS-RI system showed that all the polysaccharide samples had the similar compositions. They appeared three chromatographic peaks and the retention time were not apparently different. The Mw were 6. 570 X 10(4) g . mol-1 and 1. 393 X 10(4) g . mol-1 respectively, and one less than 10 000 which was failure to obtain accurate values. The molecular weight of the first two polysaccharide distribution index(Mw/Mn)were 1. 336 and 1. 639 respectively. The polysaccharide samples had not exhibited immunomodulatory activity assessed on the basis of nitric oxide production by RAW264. 7 macrophage cells in the experiment.


Assuntos
Frutas/química , Polissacarídeos/química , Rosaceae/química , Animais , Linhagem Celular , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Etanol , Precipitação Fracionada , Macrófagos/efeitos dos fármacos , Camundongos , Peso Molecular , Óxido Nítrico/química , Refratometria , Ácidos Sulfúricos , Água
8.
Polymers (Basel) ; 16(14)2024 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-39065289

RESUMO

A polysaccharide (CP2-S), consisting of glucose with a weight average molecular weight of 5.9 × 106, was purified from the fruit bodies of Cordyceps militaris. In this work, the corresponding structure and anti-tumor activity in vivo were investigated. Methylation and NMR analysis revealed that CP2-S was composed of a →4)-α-D-Glcp-(1→ backbone with partial substitution occurring at O-6 by T-linked α-D-Glcp in every ten residues, which has not been reported in previous reports. In vivo anti-tumor experiments showed that CP2-S could inhibit the growth of Lewis lung carcinoma in mice. Tumor inhibition rates were 17.8%, 24.5%, and 29.5% at dosages of 12.5, 50, and 100 mg/kg/d, respectively. Compared with the cisplatin group, mice treated with CP2-S exhibited a significant increase in spleen index (increased 22.7-42.4%) and thymus index (increased 47.7-36.8%). Additionally, serum levels of IgM and IgG in tumor-bearing mice increased by approximately 6.11~10.75-folds and 1.31~1.38-folds, respectively. These findings prove that CP2-S significantly inhibited the growth of Lewis lung carcinoma through immune-enhancing activity in mice.

9.
Carbohydr Polym ; 330: 121829, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38368108

RESUMO

The herbal medicine Polygonatum cyrtonema is highly regarded in China for its medicinal and dietary properties. However, further research is needed to elucidate the structure of its polysaccharide and understand how it promotes human health by modulating the gut microbiota. This study aims to investigate a homogeneous polysaccharide (PCP95-1-1) from Polygonatum cyrtonema and assess its susceptibility to digestion as well as its utilization by intestinal microbiota. The results confirmed that PCP95-1-1 is an agavin-type fructan, which possesses two fructose chains, namely ß-(2 â†’ 6) and ß-(2 â†’ 1) fructosyl-fructose, attached to the sucrose core, and has branches of ß-D-Fruf residues. Moreover, PCP95-1-1 demonstrated resistance to digestion and maintained its reducing sugar content throughout the digestive system, indicating it could reach the gut without being digested. In vitro fermentation of PCP95-1-1 significantly decreased the pH value (p < 0.05) while notably increasing the production of short-chain fatty acids (SCFAs), confirming its utilization by human gut microbiota. Additionally, PCP95-1-1 exhibited a significant ability (p < 0.05) to beneficial bacteria such as Megamonas and Bifidobacterium, while reducing the presence of facultative or conditional pathogens such as Escherichia-Shigella and Klebsiella at the genus level. Consequently, PCP95-1-1 has the potential to positively influence physical well-being by modulating the gut microbiota environment and can be developed as a functional food.


Assuntos
Microbioma Gastrointestinal , Polygonatum , Humanos , Frutanos/farmacologia , Polygonatum/química , Polissacarídeos , Frutose
10.
Int J Med Mushrooms ; 15(2): 145-52, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23557366

RESUMO

A novel small-molecule polysaccharide with a molecular mass of 2.6 kDa, was isolated from the culinary-medicinal Maitake mushroom Grifola frondosa. GFPS is composed of fucose, rhamnose, galactose, glucose, and mannose; galactose, glucose, and mannose were the dominant monosaccharides. Absorption peaks at 1077 cm-1, 1024 cm-1, and 873 cm-1, as revealed by infrared spectrum, suggesting that GFPS consists of pyranoside. GFPS significantly enhanced the production of nitric oxide and secretion of cytokines (tumor necrosis factor-α, interleukin-1ß, and interleukin-δ) from macrophages in vitro. These results indicate that this novel small-molecule polysaccharide might be beneficial for immune defense.


Assuntos
Polissacarídeos Fúngicos/química , Grifola/química , Animais , Configuração de Carboidratos , Linhagem Celular , Regulação da Expressão Gênica , Grifola/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Óxido Nítrico , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo
11.
Int J Med Mushrooms ; 15(4): 407-14, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23796222

RESUMO

To obtain a low-molecular-weight polysaccharide with immuno-enhancing activity, hot water extract of Ganoderma lucidum fruit bodies was separated by membrane ultrafiltration, anion exchange, and gel filtration chromatography, and the immunological activities of fractions were assessed on the basis of nitric oxide production by RAW 264.7 macrophages. A novel polysaccharide (TB3-2-2) was successfully isolated and purified. TB3-2-2 is a homogeneous polysaccharide, with a relative molecular weight of 5.11 × 103 Da, identified by high-performance liquid chromatography and was composed of galactose and glucose in a molar ratio of 2:3 determined by high-performance anion exchange chromatography. TB3-2-2 had a carbohydrate content of 99%, as measured using the phenol-sulfuric acid method. Proliferation of mouse spleen lymphocytes and the expression level of interleukin-6 was significantly increased by TB3-2-2. Results indicate that the low-molecular-weight polysaccharide with immunological activity in G. lucidum is worthy of further research and development.


Assuntos
Ganoderma/química , Macrófagos/efeitos dos fármacos , Polissacarídeos/química , Animais , Linhagem Celular , Regulação da Expressão Gênica/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Interleucina-6/metabolismo , Linfócitos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Baço/citologia
12.
PLoS One ; 18(9): e0291363, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37682981

RESUMO

Cordyceps militaris is a medicinal mushroom and has been extensively used as a traditional medicine in East Asia. After the chrysalis seeds are matured and harvested, the spent substrate of C. militaris still contains active ingredients but is usually discarded as waste. This study aimed to determine the antioxidant and anti-inflammatory activities of C. militaris spent substrate extract and its inhibitory activity on the Malassezia commensal yeasts that can cause dandruff and seborrheic dermatitis. Active substances in the spent substrate of C. militaris were extracted using a hot water extraction method and were used for the determination of antioxidant activity by measuring their ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radicals, hydrogen peroxide, and superoxide anions. The ability to inhibit Malassezia was analyzed using the broth microdilution method, and the reparative effect on oxidative damage in HaCaT cells was measured using in vitro cell analysis. Respiratory burst evaluation was used to determine the anti-inflammatory capacity of extracts. Analysis of the Malassezia-inhibiting activity of the extracts showed that the minimum inhibitory concentration was 6.25 mg/mL. The half maximal inhibitory concentration (IC50) values of DPPH, O2-, H2O2 and OH- were 3.845 mg/mL, 2.673 mg/mL, 0.037 mg/mL and 0.046 mg/mL, respectively. In the concentration range of 2 to 50%, the extract was non-toxic to cells and was able to protect HaCaT cells from H2O2 damage. When the volume fraction of the extract was 20.96%, its anti-inflammatory ability reached 50%. These results demonstrated that the extract may be a safe and efficacious source for pharmaceutical or cosmetic applications, with Malassezia-inhibiting, antioxidant and anti-inflammatory activities.


Assuntos
Anti-Infecciosos , Antineoplásicos , Cordyceps , Malassezia , Antioxidantes/farmacologia , Peróxido de Hidrogênio , Anti-Inflamatórios/farmacologia
13.
Int J Med Mushrooms ; 14(4): 411-7, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23510179

RESUMO

A method of detecting carbohydrates (fucose, trehalose, mannitol, arabitol, mannose, glucose, galactose, fructose, and ribose) by high-performance anion chromatography-pulsed amperometric detection (HAPEC-PAD) was established. The conditions are: CarboPac MA1 column, NaOH as the eluent, temperature 30°C, Au working electrode, Ag/AgCl reference electrode, and flow rate 0.4 mL/min. These nine analytes, which yielded high resolution by this method, could be detected in 40 minutes. Mushrooms were tested and good precision, stability, and reproducibility were achieved. This method is suitable for mushroom samples and could support research and development on sugar and sugar alcohol, which contains special effects.


Assuntos
Agaricales/química , Carboidratos/química , Cromatografia Líquida de Alta Pressão/métodos , Técnicas Eletroquímicas/métodos , Análise de Alimentos/métodos
14.
Front Nutr ; 9: 1028727, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36245525

RESUMO

To investigate the influence of molecular weight (M w) on the anti-inflammatory activity of ß-D-glucan from Ganoderma lucidum, ultrasonic irradiation was applied to treat the ß-D-glucan (GLP, 2.42 × 106 g/mol) solution to obtain two degraded fractions with molecular weight of 6.53 × 105 g/mol (GLPC) and 3.49 × 104 g/mol (GLPN). Structural analysis proved that the degraded fractions possessed similar repeated units with the original ß-D-glucan. The in vitro anti-inflammatory activity studies showed that all fractions could significantly inhibit LPS-induced expression of cytokines including TNF-α, IL-8, MIF and MCP-1 in Caco-2 cells at certain concentrations. Moreover, GLPC and GLPN exhibited better anti-inflammatory activity than GLPC. The intestinal anti-inflammatory activity evaluated by dextran sulfate sodium (DSS)-induced colitis mice model showed that intragastric administration of GLPN (lower M w fraction) could significantly recover inflamed tissues of mice. Compared with GLP and GLPC, GLPN exhibited stronger ability to inhibit the secretion of pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-6). The results revealed that M w of ß-D-glucan influenced its anti-inflammatory activity and decreasing of M w would improve the activity, which provided evidence for the potential use of ß-D-glucan from G. lucidum as anti-colitis ingredients.

15.
J Fungi (Basel) ; 8(6)2022 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-35736097

RESUMO

The effects of oleic acid addition methods on the metabolic flux distribution of ganoderic acids R, S and T's biosynthesis from Ganoderma lucidum were investigated. The results showed that adding filter-sterilized oleic acid in the process of submerged fermentation and static culture is of benefit to the synthesis of ganoderic acids R, S and T. The metabolic fluxes were increased by 97.48%, 78.42% and 43.39%, respectively. The content of ganoderic acids R, S and T were 3.11 times, 5.19 times and 1.44 times higher, respectively, than they were in the control group, which was without additional oleic acid. Ganoderic acids R, S and T's synthesis pathways (GAP), tricarboxylic acid cycles (TCA), pentose phosphate pathways (PP) and glycolysis pathways (EMP) were all enhanced in the process. Therefore, additional oleic acid can strengthen the overall metabolic flux distribution of G. lucidum in a submerged fermentation-static culture and it can reduce the accumulation of the by-product mycosterol. This study has laid an important foundation for improving the production of triterpenes in the submerged fermentation of G. lucidum.

16.
Exp Cell Res ; 316(9): 1600-9, 2010 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-20109444

RESUMO

Fibroblast migration plays an important role in the normal wound healing process; however, dysregulated cell migration may contribute to the progressive formation of fibrotic lesions in the diseased condition. To examine the role of focal-adhesion-kinase (FAK)-related non-kinase (FRNK) in regulation of fibrotic lung fibroblast migration, we examined cell migration, FRNK expression, and activation of focal adhesion kinase (FAK) and Rho GTPase (Rho and Rac) in primary lung fibroblasts derived from both idiopathic pulmonary fibrosis (IPF) patients and normal human controls. Fibrotic (IPF) lung fibroblasts have increased cell migration when compared to control human lung fibroblasts. FRNK expression is significantly reduced in IPF lung fibroblasts, while activation of FAK, Rho and Rac is increased in IPF lung fibroblasts. Endogenous FRNK expression is inversely correlated with FAK activation and cell migration rate in IPF lung fibroblasts. Forced exogenous FRNK expression abrogates the increased cell migration, and blocked the activation of FAK and Rho GTPase (Rho and Rac), in IPF lung fibroblasts. These data for the first time provide evidence that downregulation of endogenous FRNK plays a role in promoting cell migration through FAK and Rho GTPase in fibrotic IPF lung fibroblasts.


Assuntos
Movimento Celular , Fibrose Pulmonar Idiopática/metabolismo , Fibrose Pulmonar Idiopática/patologia , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/metabolismo , Western Blotting , Adesão Celular , Células Cultivadas , Regulação para Baixo , Fibroblastos/metabolismo , Fibroblastos/patologia , Imunofluorescência , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Humanos , Fibrose Pulmonar Idiopática/genética , Pulmão/metabolismo , Pulmão/patologia , Fenótipo , Proteínas Tirosina Quinases/genética , Transdução de Sinais , Proteínas rho de Ligação ao GTP/metabolismo
17.
Int J Med Mushrooms ; 13(5): 441-8, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22324409

RESUMO

The immunomodulatory effect of GLIS (Lingzhi or Reishi medicinal mushroom Ganoderma lucidum immunomodulating substance) on macrophages has been investigated as part of ongoing research into the anticancer properties of this mushroom. Proliferation of bone marrow macrophages (BMMs) was enhanced by GLIS in a dose-dependent manner. Microscopic examination revealed that numerous GLIS-treated BMMs were enlarged and formed pseudopodia. Exposure of BMMs to GLIS resulted in significant increases in NO production, induction of cellular respiratory burst activity, and increased levels of IL-1ß, IL-6, IL-12p35, IL-12p40, IL-18, and TNF-α gene expression and levels of TNF-α, IL-1ß, and IL-12 secretion. Our data indicate that GLIS activates the immune system by modulating cytokine production.


Assuntos
Células da Medula Óssea/efeitos dos fármacos , Proteoglicanas/química , Proteoglicanas/farmacologia , Reishi/química , Animais , Citocinas/genética , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Imunomodulação , Camundongos , Camundongos Endogâmicos BALB C , Explosão Respiratória/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Organismos Livres de Patógenos Específicos
18.
Int J Med Mushrooms ; 13(2): 145-52, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22135890

RESUMO

Ethanolic extracts of fruit bodies of 5 species (8 strains) of the genus Phellinus, and isolated fractions derived from 1 of these extracts (Ph. baumii PB-10), were evaluated for antioxidant activity, inhibitory effects on the growth of human tumor cells, and the capacity to protect PC12 cells against H2O2-induced oxidative damage. Extracts of all 8 strains of Phellinus spp. exhibited antioxidant activity and protected PC12 cells against oxidative damage at different magnitudes of potency. The strongest antioxidant activity was exhibited by extracts of Ph. baumii PB-10, with recorded IC50 values for superoxide radical and hydrogen peroxide scavenging activity of 3.76 microg/mL and 4.24 microg/mL, respectively. Radical-scavenging activity and protection levels against H2O2-induced damage to PC12 cells were highly correlated with the flavonoid content of the extracts and isolated fractions. All the extracts inhibited L1210, SW620, and MCF-7 tumor cell proliferation at 200 microg/mL concentrations, but inhibition was not correlated with the flavone content of the test samples and was clearly dependent upon the presence of other, as yet, unidentified components. Our data indicate that fruit bodies of species of the genus Phellinus represent a potentially valuable source of natural antioxidants of relevance to both the health and food industries.


Assuntos
Agaricales/química , Antioxidantes/farmacologia , Produtos Biológicos/farmacologia , Sequestradores de Radicais Livres/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Animais , Antioxidantes/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Flavonas/análise , Humanos , Peróxido de Hidrogênio/metabolismo , Camundongos , Neoplasias , Células PC12 , Ratos
19.
Int J Biol Macromol ; 180: 484-493, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-33689774

RESUMO

Two polysaccharide fractions (GLSB50 and GLSB70) with total sugar content of 82.07 wt% and 53.79 wt%, respectively, were obtained from the water extracts of unbroken Ganoderma lucidum spores by sequential ethanol precipitation treatment. Compared with GLSB70, GLSB50 exhibited better activity on stimulation of humoral immune responses in immunosuppressed mice. A novel ß-D-glucan (GLSB50A-III-1) with weight average molecular weight (Mw) of 1.93 × 105 g/mol was purified from GLSB50 through chromatography separation. The exponent α value of Mark-Houwink-Sakurada equation was calculated to be 0.13, indicating that GLSB50A-III-1 presented globular spheres conformation in aqueous solution. Structural analysis showed that GLSB50A-III-1 mainly consisted of (1 â†’ 3), (1 â†’ 4), (1 â†’ 6)-linked ß-d-glucose residues in the backbone, with two single ß-D-Glcp attached at O-6 of ß-(1 â†’ 3) and ß-(1 â†’ 4)-linked residues separately as side chains. The repeat unit of GLSB50A-III-1 was deduced as follows.


Assuntos
Polissacarídeos/química , Reishi/química , Esporos Fúngicos/química , beta-Glucanas/química , Animais , Sequência de Carboidratos , Cromatografia Gasosa-Espectrometria de Massas , Espectroscopia de Ressonância Magnética , Metilação , Camundongos Endogâmicos ICR , Conformação Molecular , Dados de Sequência Molecular , Estrutura Molecular , Peso Molecular , Espectroscopia de Infravermelho com Transformada de Fourier , Água/química , beta-Glucanas/isolamento & purificação
20.
Int J Med Mushrooms ; 23(3): 43-53, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33822498

RESUMO

Ganoderma lucidum mycelia are rich in active substances such as triterpenoids and sterols. However, reports on the development of effective submerged fermentation processes are lacking and the resulting total triterpene and sterol yield is still quite low. In this study, a new G. lucidum strain G0017 mycelium isolated by screening was studied in a 3-L fermenter to investigate the effect of aeration rate in liquid submerged fermentation production of triterpenoids and sterols. By fitting the specific mycelial growth rate and the specific production rate of the triterpenoid and sterol model, an effective multistage aeration rate control process for triterpenoid and sterol fermentation production was developed. This process was validated and proven in 3-L and 50-L fermenters. The resulting yields of triterpenoids and sterols were 3.34 and 3.46 g/L, respectively, which were 69.54% and 75.63% higher than the fixed aeration rate of 1.50 volume of air per volume of liquid per minute. This optimized fermentation production process conceivably could be applied to larger-scale industrial production and perhaps also to improve liquid submerged fermentation processes with relevant edible and medicinal mushrooms.


Assuntos
Agaricales/química , Reishi , Esteróis/metabolismo , Triterpenos/metabolismo , Fermentação , Micélio/crescimento & desenvolvimento
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