- L.J.L.D. Griensven van (5)
- M. Iacomini (1)
- M. Lacomini (3)
- A. Mepschen (1)
- J.J. Mes (1)
- T.R. Noordman (1)
- A.C. Ruthes (1)
- A.P. Santana-Filho (1)
- G.L. Sassaki (2)
- H.F.J. Savelkoul (1)
- R.G.M. Sman van der (1)
- F. Smiderle (1)
- F.R. Smiderle (3)
- M.Z. Tadra-Sfeir (1)
- H.A. Verhoeven (1)
- H.M. Vollebregt (1)
- H.J. Wichers (4)
Anti-inflammatory properties of the medicinal mushroom Cordyceps militaris might be related to its linear (1¿3)-ß-D-glucan.
Smiderle, F.R. ; Baggio, C.H. ; Borato, D.G. ; Santana-Filho, A.P. ; Sassaki, G.L. ; Iacomini, M. ; Griensven, L.J.L.D. van - \ 2014
PLoS One 9 (2014)10. - ISSN 1932-6203
formalin test - in-vivo - fruiting bodies - beta-glucans - congo-red - polysaccharides - mice - macrophages - extract - complex
The Ascomycete Cordyceps militaris, 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 ß-D-glucan was purified. This polymer was analysed by GC-MS and NMR spectrometry, showing a linear chain composed of ß-D-Glcp (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-1ß, TNF-a, 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 ß-(1¿3)-D-glucan, that seems to be the most potent anti-inflammatory compound present in the polysaccharide extracts of C. militaris. In vivo, ß-(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 ß-(1¿3)-D-glucan.
Phellinus linteus polysaccharide extracts increase the mitochondrial membrane potential and cause apoptotic death of THP-1 monocytes
Griensven, L.J.L.D. van; Verhoeven, H.A. - \ 2013
Chinese Medicine 8 (2013). - ISSN 1749-8546 - 13 p.
trail-induced apoptosis - cell-death - prostate-cancer - beta-glucans - tumor-growth - macrophage - necrosis - protein - fungi - differentiation
Background: The differentiation resp. death of human monocytic THP-1 cells induced by polysaccharide extracts of the medicinal mushrooms Phellinus linteus, Agaricus bisporus and Agaricus brasiliensis have been studied. This study aims to identify leads for the causal effects of these mushroom components on cell differentiation and death. Methods: THP-1 cells were treated with different polysaccharide extracts of mushrooms and controls. Morphological effects were observed by light microscopy. Flow cytometry was applied to follow the cell differentiation by cell cycle shifts after staining with propidium iodide, changes of mitochondrial membrane potential after incubation with JC-1, and occurrence of intracellular reactive oxygen species after incubation with hydroethidine. Principal component analysis of the data was performed to evaluate the cellular effects of the different treatments. Results: P. linteus polysaccharide extracts induced dose-dependent apoptosis of THP-1 cells within 24 h, while A. bisporus and A. brasiliensis polysaccharide extracts caused differentiation into macrophages. A pure P. linteus polysaccharide had no effect. Apoptosis was inhibited by preincubating THP-1 cells with human serum. The principal component analysis revealed that P. linteus, A. bisporus and A. brasiliensis polysaccharide extracts increased reactive oxygen species production. Both A. bisporus and A. brasiliensis polysaccharide extracts decreased the mitochondrial membrane potential, while this was increased by P. linteus polysaccharide extracts. Conclusions: P. linteus polysaccharide extracts caused apoptosis of THP-1 monocytes while A. bisporus and A. brasiliensis polysaccharide extracts caused these cells to differentiate into macrophages. The protective effects of human serum suggested that P. linteus polysaccharide extract induced apoptosis by extrinsic pathway, i.e. by binding to the TRAIL receptor. The mitochondrial membrane potential together with reactive oxygen species seems to play an important role in cell differentiation and cell death.
Agaricus bisporus and Agaricus brasiliensis (1 ¿ 6)-ß-d-glucans show immunostimulatory activity on human THP-1 derived macrophages
Smiderle, F.R. ; Alquini, G. ; Tadra-Sfeir, M.Z. ; Lacomini, M. ; Wichers, H.J. ; Griensven, L.J.L.D. van - \ 2013
Carbohydrate Polymers 94 (2013)1. - ISSN 0144-8617 - p. 91 - 99.
beta-glucans - fungal metabolites - molecular-weight - polysaccharides - lipopolysaccharide - mushroom - activation - receptor - rats - expression
The (1 ¿ 6)-ß-d-glucans from Agaricus bisporus and Agaricus brasiliensis were purified to evaluate their effects on the innate immune system. THP-1 macrophages were used to investigate the induction of the expression of TNF-a, IL1ß, and COX-2 by RT-PCR. The purification of the polysaccharides gave rise to fractions containing 96–98% of glucose. The samples were analyzed by GC–MS, HPSEC and 13C NMR, which confirmed the presence of homogeneous (1 ¿ 6)-ß-d-glucans. The ß-glucans were incubated with THP-1 derived macrophages, for 3 h and 6 h to evaluate their effects on the expression of pro-inflammatory genes. Both ß-glucans stimulated the expression of such genes as much as the pro-inflammatory control (LPS). When the cells were incubated with LPS + ß-glucan, a significant inhibition of the expression of IL-1ß and COX-2 was observed for both treatments after 3 h of incubation. By the results, we conclude that the (1 ¿ 6)-ß-d-glucans present an immunostimulatory activity when administered to THP-1 derived macrophages
Characterization of polarized THP-1 macrophages and polarizing ability of LPS and food compounds
Chanput, W. ; Mes, J.J. ; Savelkoul, H.F.J. ; Wichers, H.J. - \ 2013
Food & Function 4 (2013)2. - ISSN 2042-6496 - p. 266 - 276.
adipose-tissue macrophages - alternative activation - beta-glucans - mononuclear-cells - gene-expression - kappa-b - monocyte - differentiation - induction - phenotype
Little is known about the polarizing potential of currently used human macrophage cell lines, while a better understanding phenomena can support the prediction of effects in vivo based on in vitro analysis. To test the polarization capability of PMA differentiated-THP-1 macrophages (M0), cells were stimulated with 20 ng/ml IFN¿+1µg/ml LPS and 20 ng/ml IL-4, which are known to influence macrophage polarization in vivo and ex vivo into the M1 and M2 state, respectively. Apart from several well-known M1 and M2 markers, also new possible markers for M1 and M2 polarization were analysed in this study. The expression of M1 marker genes was up-regulated in IFN¿+LPS stimulated-M0 THP-1 macrophages. The IL-4 stimulated-M0 THP-1 macrophages expressed M2 cell membrane receptor genes. However, M2 chemokine and their receptor genes were only slightly up-regulated which might be due to complexity of the secondary cell-cell interaction of the chemokine system. Lipopolysaccharide from E.coli (LPS) and food compounds (lentinan, vitamin D3 (vD3) and the combination of lentinan+vitaminD3 (Len+vD3)) were investigated for their polarizing ability on M0 THP-1 macrophages towards either the M1 or M2 state. LPS (700 ng/ml) was able to skew M0 THP-1 macrophages towards the M1 direction since all analysed M1 marker genes were strongly expressed. Lentinan, vD3 and Len+vD3 did not induce expression of either M1 or M2 markers indicating no polarizing ability of these compounds. Based on the expression of M1 and M2 marker genes we concluded that THP-1 macrophages could be successfully polarized into either the M1or M2 state. Therefore, they can be used as a new macrophage polarizing model to estimate the polarizing/switching ability of test food compounds.
Review of hypotheses for fouling during beer clarification using membranes
Mepschen, A. ; Sman, R.G.M. van der; Vollebregt, H.M. ; Noordman, T.R. - \ 2012
Journal of Membrane Science 396 (2012). - ISSN 0376-7388 - p. 22 - 31.
cross-flow microfiltration - governing permeate flux - pressure-driven flow - beta-glucans - transmembrane pressure - colloidal suspensions - ceramic membranes - protein rejection - self-diffusion - model
Hypotheses concerning the fouling of membranes during beer clarification via crossflow microfiltration are reviewed. Beer has been classified into three groups of components, each having a different kind of fouling mechanisms – but also having interactions with other modes of fouling. The membrane fouling also strongly depends on the characteristics of the membrane. An optimal pore diameter and membrane morphology can be identified. The various hypotheses have been formulated in terms of mathematical models, which are tested using experimental data of dead-end filtration of beer. This comparison shows that our hypotheses are quite likely to be valid, and form a good basis for further model-based exploration of the optimization of the beer clarification process. Due to the similarity of beer with biotechnological broths, the presented fouling hypotheses extend beyond the original application of beer microfiltration.
Polysaccharides from Agaricus bisporus and Agaricus brasiliensis show similarities in their structures and their immunomodulatory effects on human monocytic THP-1 cells
Smiderle, F.R. ; Ruthes, A.C. ; Arkel, J. van; Chanput, W. ; Lacomini, M. ; Wichers, H.J. ; Griensven, L.J.L.D. van - \ 2011
BMC Complementary & alternative medicine 11 (2011). - ISSN 1472-6882 - 11 p.
mediated antitumor polysaccharides - medicinal mushroom extracts - blazei murill - fruiting body - macrophage activation - beta-glucans - tnf-alpha - himematsutake - pleurotus - mannogalactan
Background 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. Agaricus bisporus and A. brasiliensis and to study their effects on the innate immune system, in particular on the in vitro induction of pro-inflammatory cytokines, using THP-1 cells. Methods 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. Results Semi-purified polysaccharide extracts of A. bisporus and A. brasiliensis (= blazei) were found to contain (1¿6),(1¿4)-linked a-glucan, (1¿6)-linked ß-glucan, and mannogalactan. Their proportions were determined by integration of 1H-NMR signs, and were considerably different for the two species. A. brasiliensis showed a higher content of ß-glucan, while A. bisporus presented mannogalactan as its main polysaccharide. The extracts induced a comparable increase of transcription of the pro-inflammatory cytokine genes IL-1ß and TNF-a 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 A. brasiliensis extract. Conclusions The polysaccharide preparations from the closely related species A. bisporus and A. brasiliensis show major differences in composition: A. bisporus shows high mannogalactan content whereas A. brasiliensis has mostly ß-glucan. Semi-purified polysaccharide extracts from both Agaricus species stimulated the production of pro-inflammatory cytokines and enzymes, while the polysaccharide extract of A. brasiliensis reduced synthesis of these cytokines induced by LPS, suggesting programmable immunomodulation.
High molecular weight glucan of the culinary medicinal mushroom Agaricus bisporus is an a-glucan that forms complexes with low molecular weight galactan
Smiderle, F. ; Sassaki, G.L. ; Arkel, J. van; Lacomini, M. ; Wichers, H.J. ; Griensven, L.J.L.D. van - \ 2010
Molecules 15 (2010)8. - ISSN 1420-3049 - p. 5818 - 5830.
beta-glucans - flammulina-velutipes - edible mushroom - in-vitro - polysaccharide - pleurotus - extracts - glycogen - purification - spectroscopy
An a-glucan was isolated from the culinary medicinal mushroom A. bisporus by hot water extraction, ethanol precipitation and DEAE-cellulose chromatography. The resulting material showed a single HMW peak excluded from a Sephadex G50 column that could completely be degraded by a-amylase treatment. After heating in 1% SDS a small additional peak of low MW eluted from the G50 column. The monosaccharide composition of the main peak was evaluated by HPLC, and was found to consist of a majority of glucose (97.6%), and a minor proportion of galactose (2.4%). Methylation analysis and degradation by a-amylase indicated the presence of an a-glucan with a main chain consisting of (1®4)-linked units, substituted at O-6 by a-D-glucopyranose single-units in the relation 1:8. Mono- (13C-, 1H-NMR) and bidimensional [1H (obs.),13C-HSQC] spectroscopy analysis confirmed the a-configuration of the Glcp residues by low frequency resonances of C-1 at d 100.6, 100.2, and 98.8 ppm and H-1 high field ones at d 5.06, 5.11, and 4.74 ppm. The DEPT-13C-NMR allowed assigning the non-substituted and O-substituted –CH2 signals at d 60.3/60.8 and 66.2 ppm, respectively. Other assignments were attributed to C-2, C-3, C-4, C-5 and C-6 of the non-reducing ends at d 71.8; 72.8; 70.0; 71.3 and 60.3/60.8 ppm, respectively. The minor proportion of galactose that was demonstrated was probably derived from a complex between the a-glucan and a low molecular weight galactan