Polysaccharides from Agaricus bisporus and Agaricus brasiliensis show similarities in their structures and their immunomodulatory effects on human monocytic THP-1 cells

<p>Abstract</p> <p>Background</p> <p>Mushroom polysaccharides have traditionally been used for the prevention and treatment of a multitude of disorders like infectious illnesses, cancers and various autoimmune diseases. Crude mushroom extracts have been tested without detailed chemical analyses of its polysaccharide content. For the present study we decided to chemically determine the carbohydrate composition of semi-purified extracts from 2 closely related and well known basidiomycete species, i.e. <it>Agaricus bisporus </it>and <it>A. brasiliensis </it>and to study their effects on the innate immune system, in particular on the <it>in vitro </it>induction of pro-inflammatory cytokines, using THP-1 cells.</p> <p>Methods</p> <p>Mushroom polysaccharide extracts were prepared by hot water extraction and precipitation with ethanol. Their composition was analyzed by GC-MS and NMR spectroscopy. PMA activated THP-1 cells were treated with the extracts under different conditions and the production of pro-inflammatory cytokines was evaluated by qPCR.</p> <p>Results</p> <p>Semi-purified polysaccharide extracts of <it>A. bisporus </it>and <it>A. brasiliensis </it>(= <it>blazei</it>) were found to contain (1→6),(1→4)-linked α-glucan, (1→6)-linked β-glucan, and mannogalactan. Their proportions were determined by integration of ¹H-NMR signs, and were considerably different for the two species. <it>A. brasiliensis </it>showed a higher content of β-glucan, while <it>A. bisporus </it>presented mannogalactan as its main polysaccharide. The extracts induced a comparable increase of transcription of the pro-inflammatory cytokine genes IL-1β and TNF-α as well as of COX-2 in PMA differentiated THP-1 cells. Pro-inflammatory effects of bacterial LPS in this assay could be reduced significantly by the simultaneous addition of <it>A. brasiliensis </it>extract.</p> <p>Conclusions</p> <p>The polysaccharide preparations from the closely related species <it>A. bisporus </it>and <it>A. brasiliensis </it>show major differences in composition: <it>A. bisporus </it>shows high mannogalactan content whereas <it>A. brasiliensis </it>has mostly β-glucan. Semi-purified polysaccharide extracts from both <it>Agaricus </it>species stimulated the production of pro-inflammatory cytokines and enzymes, while the polysaccharide extract of <it>A. brasiliensis </it>reduced synthesis of these cytokines induced by LPS, suggesting programmable immunomodulation.</p

Caracterización bioquímica y funcional de extractos ricos en fibra dietética obtenidos de Lentinula edodes

Resumen del trabajo presentado a las III Jornadas Científicas CIAL Fórum, celebradas del 22 al 23 de noviembre de 2018 en el Instituto de Investigación en Ciencias de la Alimentación (CIAL).Las setas comestibles han formado parte de la dieta desde hace siglos y sus propiedades nutricionales y beneficiosas para la salud han estimulado un crecimiento exponencial de su consumo en las últimas décadas. En particular, la seta shiitake (Lentinula edodes) es una de las más valoradas, no solo por sus características culinarias sino también por su contenido en compuestos bioactivos. La obtención de la fracción rica en fibra dietética se realizó sometiendo a cuerpos fructíferos de shiitake pulverizados a elevadas temperaturas (98ºC) en medio acuoso para eliminar compuestos hidrosolubles y concentrar aquellos insolubles, tales como polímeros de difícil degradación durante la digestión humana, que suelen ser fermentados por el microbioma del colon, es decir, actúan como fibra dietética. El extracto obtenido era rico en quitinas y ciertos tipos de β-glucanos y también contenía ergosterol, un lípido que reduce la absorción de colesterol y puede dar lugar a vitamina D2 mediante la irradiación con luz ultravioleta. La quitina presente puede ser sometida a protocolos de desacetilación para dar lugar a quitosano, polímero bien estudiado por sus propiedades hipolipidémicas, antimicrobianas y antitumorales. En este trabajo, además de su caracterización bioquímica, se llevó a cabo la evaluación de su capacidad antioxidante e hipocolesterolémica y sus propiedades reológicas. Además, se evaluaron diferentes condiciones para elaborar un protocolo de irradiación con luz ultravioleta para la conversión de ergosterol en vitamina D2 y de desacetilación química para la conversión de quitina en quitosano.Los autores agradecen la financiación otorgada por el Ministerio de Ciencia, Innovación y Universidades a través del proyecto AGL2014-56211-R y a la Comunidad de Madrid (S2013/ABI-2728).Peer reviewe

Vitamin D-enriched extracts obtained from shiitake mushrooms (Lentinula edodes) by supercritical fluid extraction and UV-irradiation

The combination of supercritical fluid extraction followed by UV-irradiation resulted in an interesting strategy to obtain ergosterol- and Vitamin D2-enriched extracts from Lentinula edodes to design novel functional food. Extractions carried out following a specific experimental design pointed out that extraction yields and ergosterol concentrations were more influenced by pressure than by the extraction temperature although, high temperatures (55–75 °C) might induce transformation of ergosta-7,22-dienol and fungisterol into ergosterol. After extraction, the ergosterol-enriched extracts should be dissolved in methanol or ethanol and irradiated (25 °C, 4 cm) to partially transform ergosterol into vitamin D2. Irradiation at 365 nm was less effective than at 254 nm. The fastest transformation was obtained using a UV lamp covering the complete UV spectrum for a maximum of 1 h. However, this lamp also induced vitamin D4 formation although in lower amounts than vitamin D2 or lumisterol2 while with irradiation at 254 nm most of ergosterol was transformed into vitamin D2.This research was supported by national R + D program from the Spanish Ministry of Science and Innovation (project AGL2014-56211-R) and the regional program from the Community of Madrid, Spain (S2013/ABI-2728). F.R.S. thanks to the Brazilian funding agency CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico).Peer reviewe

Water-soluble polysaccharide extracts from the oyster culinary-medicinal mushroom pleurotus ostreatus (Agaricomycetes) with HMGCR inhibitory activity

Water extracts from Pleurotus ostreatus containing no statins showed 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR) inhibitory activity (in vitro) that might be due to specific water-soluble polysaccharides (WSPs); when isolated and deproteinized, increasing concentrations of the WSP extract induced higher inhibition. The WSP extract contained mainly β-glucans, mannogalactans, and glycogen (e.g., α-glucans), although derivatives or fragments with lower molecular weights (between 14 and 3.5 kDa) were present and were able to induce the inhibitory activity. The extract contained more β-(1→3)-glucans than β-(1→3),(1→6)-glucans, and they partially survived digestion and managed to pass through Caco2 cell monolayers to the lower compartment after in vitro digestion and transport experiments. The WSP might also modulate Caco2 membrane integrity

Simple and effective purification approach to dissociate mixed water-insoluble α- and β-D-glucans and its application on the medicinal mushroom Fomitopsis betulina

Differences in anomericity and in the branching degree of glucans lead to characteristic intermolecular association that influences their solubility in water or other solvents. A simple purification approach, based on the glucan solubility in aq. 0.1 M NaOH solution, was applied for the separation of mixed water-insoluble α-D-glucans from β-D-glucans extracted from fruiting bodies of Fomitopsis betulina, which is an underexploited medicinal mushroom. The results indicated that the β-D-glucan is constituted by (1→3)-linked β-D-Glcp units substituted at O-6 by non-reducing β-D-Glcp and (1→6)-linked β-D-Glcp units, while the α-D-glucan has a linear (1→3)-linked glucan structure. Thus, the 0.1 M NaOH treatment proved to be a simple, efficient and low-cost purification method for separation of water-insoluble glucans with different anomeric configurations and degree of branching that were interacting by intermolecular forces

Simple and effective purification approach to dissociate mixed water-insoluble α- and β-D-glucans and its application on the medicinal mushroom Fomitopsis betulina

Differences in anomericity and in the branching degree of glucans lead to characteristic intermolecular association that influences their solubility in water or other solvents. A simple purification approach, based on the glucan solubility in aq. 0.1 M NaOH solution, was applied for the separation of mixed water-insoluble α-D-glucans from β-D-glucans extracted from fruiting bodies of Fomitopsis betulina, which is an underexploited medicinal mushroom. The results indicated that the β-D-glucan is constituted by (1→3)-linked β-D-Glcp units substituted at O-6 by non-reducing β-D-Glcp and (1→6)-linked β-D-Glcp units, while the α-D-glucan has a linear (1→3)-linked glucan structure. Thus, the 0.1 M NaOH treatment proved to be a simple, efficient and low-cost purification method for separation of water-insoluble glucans with different anomeric configurations and degree of branching that were interacting by intermolecular forces.</p

Phytochemical Evaluation and Anti-Inflammatory Potential of <i>Miconia albicans</i> (Sw.) Triana Extracts

The plant Miconia albicans (Sw.) Triana has been popularly used in Brazil to treat chronic inflammatory disturbances, such as osteoarthritis. This disease affects 250 million people worldwide, and is associated with intense pain and loss of articular function. There is a lack of information about the phytochemistry and bioactivity of M. albicans. Therefore, this study determined the chemical composition of some extracts and evaluated their cytotoxicity, along with their antioxidant and anti-inflammatory, activities using in vitro models. Aqueous and ethanolic extracts were prepared. Afterwards, a liquid–liquid partition was developed using chloroform, ethyl acetate, and n-butanol. The extracts were characterized by LC–MS, and their biological activities were evaluated on epithelial cells (Vero), tumoral hepatic cells (Hep-G2), and THP-1 macrophages. LC–MS analyses identified several flavonoids in all fractions, such as quercetin, myricetin, and their glycosides. The crude extracts and n-butanol fractions did not present cytotoxicity to the cells. The non-toxic fractions presented significant antioxidant activity when evaluated in terms of DPPH scavenging activity, lipid peroxidation, and ROS inhibition. THP-1 macrophages treated with the n-butanol fraction (250 µg/mL) released fewer pro-inflammatory cytokines, even in the presence of LPS. In the future, it will be necessary to identify the phytochemicals that are responsible for anti-inflammatory effects for the discovery of new drugs. In vivo studies on M. albicans extracts are still required to confirm their possible mechanisms of action

Chemical characterization and wound healing property of a β-D-glucan from edible mushroom Piptoporus betulinus

A water-soluble β-D-glucan was obtained from fruiting bodies of Piptoporus betulinus, by hot aqueous extraction followed by freeze-thawing procedure and dialysis. Its molar mass distribution and conformational behavior in solution was assessed by size-exclusion chromatography coupled with multiangle laser light scattering, showing a polysaccharide with an average molecular weight of 2.5 × 105 Da with a random coil conformation for molecular weights below 1 × 106 Da. Typical signals of β-(1 → 3)-linkages were observed in NMR spectrum (δ 102.7/4.76; 102.8/4.74; 102.9/4.52; and δ 85.1/3.78; 85.0/3.77) and also signals of O-6 substitution at δ 69.2/4.22 and 69.2/3.87. The analysis of partially O-methylated alditol acetates corroborates the NMR results, indicating the presence of a β-D-glucan with a main chain (1 → 3)-linked, substituted at O-6 by single-units of glucose. The β-D-glucan showed no toxicity on human colon carcinoma cell line (Caco-2) up to 1000 μg mL-1 and promoted cell migration on in vitro scratch assay, demonstrating a potential wound healing capacity

Chemical characterization and wound healing property of a β-D-glucan from edible mushroom Piptoporus betulinus

A water-soluble β-D-glucan was obtained from fruiting bodies of Piptoporus betulinus, by hot aqueous extraction followed by freeze-thawing procedure and dialysis. Its molar mass distribution and conformational behavior in solution was assessed by size-exclusion chromatography coupled with multiangle laser light scattering, showing a polysaccharide with an average molecular weight of 2.5 × 105 Da with a random coil conformation for molecular weights below 1 × 106 Da. Typical signals of β-(1 → 3)-linkages were observed in NMR spectrum (δ 102.7/4.76; 102.8/4.74; 102.9/4.52; and δ 85.1/3.78; 85.0/3.77) and also signals of O-6 substitution at δ 69.2/4.22 and 69.2/3.87. The analysis of partially O-methylated alditol acetates corroborates the NMR results, indicating the presence of a β-D-glucan with a main chain (1 → 3)-linked, substituted at O-6 by single-units of glucose. The β-D-glucan showed no toxicity on human colon carcinoma cell line (Caco-2) up to 1000 μg mL-1 and promoted cell migration on in vitro scratch assay, demonstrating a potential wound healing capacity

Anti-Inflammatory Properties of the Medicinal Mushroom <i>Cordyceps militaris</i> Might Be Related to Its Linear (1→3)-β-D-Glucan

<div><p>The Ascomycete <i>Cordyceps militaris</i>, an entomopathogenic fungus, is one of the most important traditional Chinese medicines. Studies related to its pharmacological properties suggest that this mushroom can exert interesting biological activities. Aqueous (CW and HW) and alkaline (K5) extracts containing polysaccharides were prepared from this mushroom, and a β-D-glucan was purified. This polymer was analysed by GC-MS and NMR spectrometry, showing a linear chain composed of β-D-Glc<i>p</i> (1→3)-linked. The six main signals in the ¹³C-NMR spectrum were assigned by comparison to reported data. The aqueous (CW, HW) extracts stimulated the expression of IL-1β, TNF-α, and COX-2 by THP-1 macrophages, while the alkaline (K5) extract did not show any effect. However, when the extracts were added to the cells in the presence of LPS, K5 showed the highest inhibition of the pro-inflammatory genes expression. This inhibitory effect was also observed for the purified β-(1→3)-D-glucan, that seems to be the most potent anti-inflammatory compound present in the polysaccharide extracts of <i>C. militaris. In vivo</i>, β-(1→3)-D-glucan also inhibited significantly the inflammatory phase of formalin-induced nociceptive response, and, in addition, it reduced the migration of total leukocytes but not the neutrophils induced by LPS. In conclusion, this study clearly demonstrates the anti-inflammatory effect of β-(1→3)-D-glucan.</p></div