RESUMO
This study aimed to assess the effects of polysaccharides extracted from Hericium erinaceus fruiting bodies (HEFPs) on the inflammatory response to oxidative stress in a mouse model of ulcerative colitis (UC) induced by ingestion of dextran sodium sulfate. The results indicated reduced oxidative damage in the HEFPs groups, as evidenced by significantly decreased malondialdehyde levels and significantly increased levels of the antioxidant enzymes superoxide dismutase and catalase in colon homogenates, compared with those in the Model Control (MC) group. Additionally, compared with the levels in the MC group, the levels of the pro-inflammatory factors IL-6, IL-1ß, and TNF-α in the positive-control (PC) and HEFPs groups were significantly lower, and that of the anti-inflammatory factor IL-10 was significantly higher. qRT-PCR analyses revealed that the colon expression patterns of IL-6, IL-1ß, TNF-α, and IL-18 were consistent with the serum levels. Western-blotting results indicated significantly lower levels of NLRP3, ASC, and caspase 1 P20 in the HEFPs and PC groups than in the MC group. These findings suggest that HEFPs alleviate UC by suppressing the NLRP3 inflammasome/Caspase-1 pathway. Lachnospiraceae, Clostridiales, Parabacteroides, Oscillibacter, and Clostridium XlVa genera were more abundant in the gut microbiota of the HEFPs group than that of the MC group.
Assuntos
Colite Ulcerativa , Hericium , Inflamassomos , Proteína 3 que Contém Domínio de Pirina da Família NLR , Animais , Colite Ulcerativa/tratamento farmacológico , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Camundongos , Inflamassomos/metabolismo , Inflamassomos/efeitos dos fármacos , Hericium/química , Masculino , Homeostase/efeitos dos fármacos , Polissacarídeos Fúngicos/farmacologia , Polissacarídeos Fúngicos/química , Modelos Animais de Doenças , Polissacarídeos/farmacologia , Polissacarídeos/química , Sulfato de Dextrana , Estresse Oxidativo/efeitos dos fármacos , Citocinas/metabolismo , Intestinos/efeitos dos fármacosRESUMO
Lentinula edodes has high nutritional value and abundant protein. In order to develop and utilize edible mushroom protein, this study was designed to investigate the effects of TGase-catalyzed glycosylation and cross-linking on the physicochemical and functional properties of Lentinus edodes protein fraction. The results showed that within a certain time, glycosylation and TGase-catalyzed glycosylation decreased the total sulfydryl, free sulfydryl, disulfide bond, surface hydrophobicity, ß-fold and α-helix, but increased the fluorescence intensity, random coil, ß-turn, particle size and thermal stability. The apparent viscosity and the shear stress of the protein with an increase in shear rate were increased, indicating that TGase-catalyzed glycosylation promoted the generation of cross-linked polymers. In addition, the TGase-catalyzed glycosylated proteins showed a compact texture structure similar to the glycosylated proteins at the beginning, indicating that they formed a stable three-dimensional network structure. The flaky structure of proteins became more and more obvious with time. Moreover, the solubility, emulsification, stability and oil-holding capacity of enzymatic glycosylated Lentinus edodes protein fraction were significantly improved because of the proper TGase effects of glycosylation grafting and cross-linking. These results showed that glycosylation and TGase-catalyzed glycosylation could improve the processing characteristics of the Lentinula edodes protein fraction to varying degrees.
RESUMO
Dietary supplements have improved the prevention of insulin resistance and metabolic diseases, which became a research hotspot in food science and nutrition. Obesity and insulin resistance, caused by a high-fat diet, eventually result in severe metabolic diseases, can be prevented with the dietary supplement D-chiro-inositol (DCI). In this work, we isolated mice primary hepatocytes with palmitic acid stimulation and DCI was applied to compare and contrast its effects of in primary hepatocyte biology. Before and after intervention with DCI, we used RNA-Seq technology to establish a primary hepatocyte transcriptome gene profile. We found that both PA and DCI cause a wide variation in gene expression. Particularly, we found that DCI plays critical role in this model by acting on glycolysis and gluconeogenesis. Overall, we generated extensive transcripts from primary hepatocytes and uncovered new functions and gene targets for DCI.
Assuntos
Biomarcadores/sangue , Suplementos Nutricionais , Regulação da Expressão Gênica/efeitos dos fármacos , Hepatócitos/metabolismo , Inositol/farmacologia , Resistência à Insulina , Ácido Palmítico/toxicidade , Animais , Inibidores Enzimáticos/toxicidade , Gluconeogênese , Glicólise , Hepatócitos/efeitos dos fármacos , Camundongos , RNA-Seq , Complexo Vitamínico B/farmacologiaRESUMO
The Glistening Inkcap (Coprinellus micaceus) is a wild saprobic mushroom in China. In this study, we assembled and annotated its complete mitochondrial genome using raw data sequenced through Illumina NovaSeq 6000 platform (Illumina, San Diego, CA). The length of the C. micaceus mitochondrial genome is 65,450 bp with 33.05% GC content. Totally, 41 genes, including 14 protein-coding genes, 25 tRNAs and 2 rRNAs were identified in the mitochondrial genome. Phylogenetic analysis showed that the mitochondrial genome relationship between C. micaceus and Coprinopsis cinerea was the closest.
RESUMO
The fruiting bodies of Hericium erinaceus (Bull.) Pers. are commonly used in China in the treatment of digestive system diseases. In this work, the polysaccharides from the fruiting bodies of Hericium erinaceus (HEFPs) were extracted, and their effects on human colorectal cancer cells (HCT-116 and DLD1) were investigated in vitro. Our results showed that HEFPs were mainly composed of arabinose, galactose, glucose, and mannose at a molar ratio of 8.99 : 11.15 : 1.2 : 1.97. They significantly inhibited the growth of these cells by inducing apoptosis by the modulation of Bax and Bcl-2 expression, which in turn induced the loss of mitochondrial membrane potential, leading to the activation of cleaved-caspase-9 and cleaved-caspase-3. These results suggested that HEFPs induced apoptosis via the caspase-9-depedent intrinsic mitochondrial pathway. Furthermore, HEFPs increased the level of reactive oxygen species (ROS) in HCT-116 and DLD1 cells. The addition of the antioxidant N-acetyl-l-cysteine reduced the ability of HEFPs to trigger the intrinsic mitochondrial pathway, indicating the role of ROS generation in the upstream pathway of HEFP-induced apoptosis. Therefore, the results described in this study could be of interest for further studies in finding functional foods or alternative therapeutic agents against colorectal cancer.
Assuntos
Apoptose/efeitos dos fármacos , Hericium/metabolismo , Polissacarídeos/farmacologia , Transdução de Sinais , Acetilcisteína/metabolismo , Agaricales/química , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Neoplasias Colorretais/tratamento farmacológico , Células HCT116 , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Polissacarídeos/análise , Espécies Reativas de Oxigênio/metabolismoRESUMO
The fruiting body of Hericium erinaceus has been used to treat digestive system disorder-related diseases for over 2000 years in China. A novel polysaccharide, HEFP-2b, was obtained from H. erinaceus fruiting bodies. Physical and chemical analysis showed that HEFP-2b consisted of fucose, galactose, glucose, and mannose in molar ratio of 11.81:22.82:44.28:21.09, and that its molecular weight was 3.252 × 104 Da. The backbone of HEFP-2b consisted of â6)-linked-α-D-Glcp-(1â and â4)-ß-D-Galp-(1â and â3,6) -α-D-Manp linkage, with two side-branching units of (1â and â6)-ß-D-Galp and (1â and â4)-α-D-Manp, terminated by Glc and Fuc. The results of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cell cycle arrest experiments revealed that HEFP-2b considerably inhibited the growth of colon cancer cells (HCT-116) in vitro. The growth inhibitory effects of HEFP-2b correlated with their ability to arrest the cell cycle at the S-phase. Our results will provide valuable information for future studies on HEFP-2b as a novel health-promoting functional food ingredient that can be used for treating colon cancer.
Assuntos
Carpóforos/química , Hericium/química , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Pontos de Checagem da Fase S do Ciclo Celular/efeitos dos fármacos , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular , Neoplasias do Colo , Relação Dose-Resposta a Droga , Células HCT116 , Humanos , Monossacarídeos/análiseRESUMO
A novel 57-kDa acidic α-galactosidase designated as HEG has been purified from the dry fruiting bodies of Hericium erinaceus. The isolation protocol involved ion-exchange chromatography and gel filtration on a Superdex75 column. The purification fold and specific activity were 1251 and 46â¯units/mg, respectively. A BLAST search of internal peptide sequences obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis suggested that the enzyme belonged to the GH27 family. The activity of the enzyme reached its maximum at a pH of 6.0 or at 60⯰C. The enzyme was stable within an acidic pH range of 2.2-7.0 and in a narrow temperature range. The enzyme was strongly inhibited by Zn2+, Fe3+, Ag+ ions and SDS. The Lineweaver-Burk plot suggested that the mode of inhibition by galactose and melibiose were of a mixed type. N-bromosuccinimide drastically decreased the activity of the enzyme, whereas diethylpyrocarbonate and carbodiimide strengthened the activity slightly. Moreover, the isolated enzyme displayed remarkable resistance to acid proteases, neutral proteases and pepsin. The enzyme could also hydrolyse oligosaccharides and polysaccharides. In addition, acidic protease promoted the hydrolysis of RFOs by HEG. The Km values of the enzyme towards pNPGal, raffinose and stachyose were 0.36â¯mM, 40.07â¯mM and 54.71â¯mM, respectively. These favourable properties increase the potential of the enzyme in the food industry and animal feed applications.
Assuntos
Basidiomycota/enzimologia , Peptídeo Hidrolases/metabolismo , alfa-Galactosidase/isolamento & purificação , alfa-Galactosidase/metabolismo , Sequência de Aminoácidos , Inibidores Enzimáticos/farmacologia , Hidrólise , Cinética , Peso Molecular , Rafinose/metabolismo , Especificidade por Substrato , alfa-Galactosidase/antagonistas & inibidores , alfa-Galactosidase/químicaRESUMO
In some industrial processes, cold and light stresses are recognized as two important environmental triggers for the transformation of mycelia into fruit-bodies via intermediate primordia in Flammulina velutipes cultivation. To gain insights into the mechanism of regulation of F. velutipes mycelia in response to cold and light stress, proteins expressed abundantly and characteristically at particular stress states were investigated by using the isobaric tags for the relative and absolute quantitation labeling technique. Among the 1046 nonredundant proteins identified with a high degree of confidence, 264 proteins, which were detected as differentially expressed proteins, were associated with 176 specific KEGG pathways. In-depth data analysis revealed that the regulatory network underlying the cold and light response mechanisms of F. velutipes mycelia was complex and multifaceted, as it included varied functions such as rapid energy supply, the biosynthesis of lysine, phenylalanine, tyrosine, and γ-aminobutyric acid, the calcium signal transduction process, dynein-dependent actin and microtubule cytoskeleton formation, autolysis, oxidative stress adaptation, pigment secretion, tissue and organ morphogenesis, and other interesting stress-related processes. Insights into the proteins might shed light on an intuitive understanding of the cold and light stress response mechanism underlying the fruiting processes of F. velutipes. Furthermore, the data might also provide further insights into the stress response mechanism of macro-fungi and valuable information for scientific improvement of some mushroom cultivation techniques in practice.
Assuntos
Flammulina/efeitos da radiação , Proteínas Fúngicas/química , Micélio/metabolismo , Proteoma/química , Proteômica/métodos , Cromatografia Líquida de Alta Pressão , Flammulina/química , Flammulina/genética , Flammulina/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Luz , Espectrometria de Massas , Micélio/química , Micélio/genética , Micélio/efeitos da radiação , Proteoma/genética , Proteoma/metabolismo , TemperaturaRESUMO
Understanding the molecular mechanisms regulating the fruiting process in macro-fungi, especially industrially cultivated mushrooms, has long been a goal in mycological research. To gain insights into the events accompanying the transformation of mycelia into fruit-bodies in Flammulina velutipes, proteins expressed characteristically and abundantly at primordium and fruit-body stages were investigated by using the iTRAQ labeling technique. Among the 171 differentially expressed proteins, a total of 68 displayed up-regulated expression levels that were associated with 84 specific KEGG pathways. Some up-regulated proteins, such as pyruvate carboxylase, aldehyde dehydrogenase, fatty acid synthase, aspartate aminotransferase, 2-cysteine peroxiredoxin, FDS protein, translation elongation factor 1-alpha, mitogen-activated protein kinases (MAPKs), and heat-shock protein 70 that are involved in carbohydrate metabolism, carotenoid formation, the TCA cycle, MAPK signaling pathway, and the biosynthesis of fatty acids and branched-chain amino acids, could serve as potential stage-specific biomarkers to study the fruiting process in F. velutipes. Knowledge of the proteins might provide valuable evidence to better understand the molecular mechanisms of fruit-body initiation and development in basidiomycete fungi. Furthermore, this study also offers valuable evidence for yield improvement and quality control of super golden-needle mushroom in practice.
Assuntos
Flammulina/metabolismo , Carpóforos/crescimento & desenvolvimento , Proteínas Fúngicas/metabolismo , Proteoma/metabolismo , Flammulina/química , Flammulina/genética , Flammulina/crescimento & desenvolvimento , Carpóforos/química , Carpóforos/genética , Carpóforos/metabolismo , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , Micélio/química , Micélio/genética , Micélio/metabolismo , Proteoma/química , Proteoma/genéticaRESUMO
Temperature is one of the pivotal factors influencing mycelium growth and fruit-body formation of Flammulina velutipes. To gain insights into hyphae growth and fruit-body formation events and facilitate the identification of potential stage-specific biomarker candidates, we investigated the proteome response of F. velutipes mycelia to cold stresses using iTRAQ-coupled two-dimensional liquid chromatography tandem mass spectrometry (2D LC-MS/MS) technique. Among 1198 proteins identified with high confidence, a total of 63 displayed altered expression level after cold stress treatments. In-depth data analysis reveals that differentially expressed proteins were involved in a variety of cellular processes, particularly metabolic processes. Among the 31 up-regulated proteins, 24 (77.42%) were associated with 22 specific KEGG pathways. These up-regulated proteins could possibly serve as potential biomarkers to study the molecular mechanisms of F. velutipes mycelia response to cold stresses. These data of the proteins might provide valuable evidences to better understand the molecular mechanisms of mycelium resistance to cold stress and fruit-body formation in fungi. BIOLOGICAL SIGNIFICANCE: Low-temperature is one of the pivotal factors in some Flammulina velutipes industrial processes influencing mycelium growth, inducing primordia and controlling fruit-body development. Preliminary study has indicated that effectively regulating cultivation could augment the yield by controlling optimal cold stress level on mycelia. However, we are still far from understanding the molecular and physiological mechanisms of adaptation of these fungi at cold stress. In the present study, the experiments reported above were undertaken to investigate chronological changes of protein expression during F. velutipes mycelia in response to cold stress by using iTRAQ-coupled 2D LC-MS/MS technique. This result would provide new insights to the underlying mycelium growth and fruit-body formation mechanisms of basidiomycetes under cold stress.
Assuntos
Temperatura Baixa , Flammulina/química , Proteoma/análise , Proteômica/métodos , Estresse Fisiológico , Cromatografia Líquida , Flammulina/crescimento & desenvolvimento , Flammulina/fisiologia , Proteínas Fúngicas/análise , Regulação Fúngica da Expressão Gênica , Estágios do Ciclo de Vida , Micélio/crescimento & desenvolvimentoRESUMO
Flammulina velutipes is a potentially excellent fungus to study basic mechanisms of basidiomycete mycelium biology. To provide a better understanding of the mechanism of hyphae growth and fruit-body formation, the biological functions of the differentially abundant proteins between the fruiting dikaryon and the non-fruiting monokaryon of F. velutipes were investigated at the proteomic level using iTRAQ-coupled two-dimensional liquid chromatography tandem mass spectrometry technique. Among the 1198 proteins identified with high confidence, a total of 472 proteins were detected differentially abundant at least one of the mycelium development stages. In-depth data analysis revealed that differentially expressed proteins were influenced a variety of cellular processes, particularly metabolic processes. Functional pathway analysis indicated that 63 up-regulated proteins at only the fruiting dikaryon (Fv13) stage were mainly distributed in 51 specific Kyoto Encyclopedia of Genes and Genome pathways, such as amino acids biosynthesis and metabolism, signaling pathway, and central carbon metabolism. These up-regulated proteins could possibly serve as potential biomarkers to study the mycelium development pathways as well as provide new insights on the mycelium heterogenic compatibility and fruit-body formation mechanisms of basidiomycetes.
Assuntos
Flammulina/crescimento & desenvolvimento , Carpóforos/crescimento & desenvolvimento , Proteínas Fúngicas/química , Cromatografia Líquida/métodos , Flammulina/química , Flammulina/genética , Flammulina/metabolismo , Carpóforos/química , Carpóforos/genética , Carpóforos/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Hifas/química , Hifas/genética , Hifas/crescimento & desenvolvimento , Hifas/metabolismo , Micélio/química , Micélio/genética , Micélio/crescimento & desenvolvimento , Micélio/metabolismo , Proteômica/métodos , Espectrometria de Massas em Tandem/métodosRESUMO
To evaluate the immune activation and reactive oxygen species scavenging activity of Cordyceps militaris polysaccharides (CMP) in vivo, 24 male and 24 female Kunming mice were randomly divided into four groups. The mice in the four experimental groups were administered 0 (normal control), 50, 100, or 200mg/kg/d body weight CMP via gavage. After 30 days, the viscera index, leukocyte count, differential leukocyte count, immunoglobulin (IgG) levels, and biochemical parameters were measured. The effect of CMP on the expression of tumor necrosis (TNF)-α, interferon (IFN)-γ, and interleukin (IL)-1ß in the spleens of experimental mice was investigated by real-time polymerase chain reaction. The results showed that the administration of CMP improved the immune function in mice, significantly increased the spleen and thymus indices, the spleen lymphocyte activity, the total quantity of white blood cells, and IgG function in mice serum. CMP exhibited significant antioxidative activity in mice, and decreased malondialdehyde levels in vivo. CMP upregulated the expression of TNF-α, IFN-γ, and IL-1ß mRNA in high-dose groups compared to that observed for the control mice. We can thus conclude that CMP effectively improved the immune function through protection against oxidative stress. CMP thus shows potential for development as drugs and health supplements.