The inhibitory effect of oral administration of lentinan on DSS-induced inflammation is exerted by the migration of T cells activated in the ileum to the colon.

Oral administration of lentinan ameliorated dextran sulfate sodium (DSS)-induced colitis through Dectin-1 receptor on intestinal epithelial cells. However, it is unclear where lentinan affects in the intestine to prevent the inflammation. We found that the administration of lentinan has induced migration of CD4+ cells from the ileum to the colon by using Kikume Green-Red (KikGR) mice in this study. This result suggests that the oral lentinan treatment could accelerate the migration of Th cells in lymphocyte from ileum into the colon during lentinan intake. Then, C57BL/6 mice were administered 2% DSS to induce colitis. The mice were administered lentinan daily via oral or rectal route before DSS administration. Its rectal administration also suppressed DSS-induced colitis, but its suppressive effects were lower compared to when orally administered, indicating that the biological responses to lentinan in the small intestine contributed to the anti-inflammatory effects. In normal mice (without DSS treatment), the expression of Il12b was significantly increased in the ileum by the oral administration of lentinan, but not by rectal one. On the other hand, no change was observed in the colon by either administration method. In addition, Tbx21 was significantly increased in the ileum. These suggested that IL-12 was increased in the ileum and Th1 cells differentiated in dependence on it. Therefore, Th1 predominant condition in the ileum could influence immunity in the colon and improve the colitis.

Anti-Allergic Activity of Fucoidan Can Be Enhanced by Coexistence with Quercetin.

In recent years, the incidence of type I hypersensitivity including hay fever has been increasing year by year in Japan. Our previous study using mice showed that only oral, but not intraperitoneal, administration of fucoidan extracted from seaweed (Saccharina japonica) suppressed type I hypersensitivity by secretion of galectin-9, which has a high affinity for IgE in the blood. However, the amount of seaweed required to achieve this activity is quite high (12 g dry weight per person per day). Therefore, the purpose of this study was to search for food ingredients in vegetables that enhance type I hypersensitivity suppression effect when consumed together with fucoidan. As a result, the enhanced effect was observed in extracts from Welsh onions and onions among vegetables. When we compared the polyphenols in the vegetables that showed activity with those that did not, flavonols such as quercetin and kaempferol were found as candidates. When quercetin or kaempferol (100 µg each) were orally administered at the same time, even at amounts where fucoidan alone showed no anti-allergic activity, anti-allergic effects were observed. More interestingly, when both flavonols were combined and administered simultaneously at half the amount of each of the above flavonols (50 µg), while the fucoidan amount remained the same, a similar effect was observed as when each flavonol (100 µg) was administered alone. The simultaneous intake of fucoidan and vegetables containing high contents of quercetin or kaempferol may reduce fucoidan intake while maintaining the allergy suppression effect, suggesting the importance of food combination.

Anti-allergic property of 4,8-sphingadienine stereoisomers in vivo and in vitro model.

4,8-Sphingadienines (SD), metabolites of glucosylceramides (GlcCer), are sometimes determined as key mediators of the biological activity of dietary GlcCer, and cis/trans geometries of 4,8-SD have been reported to affect its activity. Since regulating excessive activation of mast cells seems an important way to ameliorate allergic diseases, this study was focused on cis/trans stereoisomeric-dependent inhibitory effects of 4,8-SD on mast cell activation. Degranulation of RBL-2H3 was inhibited by treatment of 4-cis-8-trans- and 4-cis-8-cis-SD, and their intradermal administrations ameliorated ear edema in passive cutaneous anaphylaxis reaction, but 4-trans-8-trans- and 4-trans-8-cis-SD did not. Although the activation of mast cells depends on the bound IgE contents, those stereoisomers did not affect IgE contents on RBL-2H3 cells after the sensitization of anti-TNP IgE. These results indicated that 4-cis-8-trans- and 4-cis-8-cis-SD directly inhibit the activation of mast cells. In conclusion, it was assumed that 4,8-SD stereoisomers with cis double bond at C4-position shows anti-allergic activity by inhibiting downstream pathway after activation by the binding of IgE to mast cells.

Pleurotus citrinopileatus polysaccharide stimulates anti-inflammatory properties during monocyte-to-macrophage differentiation.

Polysaccharides from edible mushrooms possess important immunomodulating effects on immune cells including monocytes and macrophages. Macrophages activated by LPS/IFNγ are polarized toward inflammatory macrophages, whereas the anti-inflammatory properties of alternative activated macrophages play an important regulatory role in the innate immune system. We here show that the Pleurotus citrinopileatus mushroom polysaccharide (PCPS) can modulate the monocyte-to-macrophage differentiation early at the monocyte stage. Using both human THP-1 monocytic cells as well as human peripheral monocytes, we showed that PCPS inhibits the secreted levels of the pro-inflammatory cytokines TNF and IL-6, after stimulation of macrophages derived from PCPS-treated monocytes, with IFNγ + LPS. In addition, the glucan induced a tendency to increase the secreted levels of the anti-inflammatory cytokine IL-10, enhanced the expression levels of CCL2 and CCL8 mRNAs, and inhibited expression of CCR2 mRNA in the IFNγ/LPS activated macrophages. Interestingly, these data suggest that PCPS can induce a long-lasting anti-inflammatory effect in monocytes. Treatment of monocytes with laminarin and antibodies against Dectin-1 and TLR2 during PCPS treatment affected the glucan-modulated macrophage differentiation. In summary, the results of this study indicate that the glucan directs the differentiation of monocytes toward a macrophage cell population with reduced pro-inflammatory capacity via Dectin-1 and TLR2.

A Proinflammatory Effect of the ß-Glucan from Pleurotus cornucopiae Mushroom on Macrophage Action.

PCPS from P. citrinopileatus mushroom extract is a ß-1,6-glucan possessing a proinflammatory effect on innate immune cells. The PCPS stimulated THP-1 macrophages to secrete significant levels of TNF. Moreover, the mRNA expressions of TNF and IL-1ß were significantly enhanced by PCPS treatment. However, the PCPS did not induce to express both IL-12 and IL-10 mRNA in the macrophages. Next, the P. cornucopiae extract (containing mainly PCPS) treatment against mice showed significant increases in TNF and IL-1ß mRNA expressions in the peritoneal macrophages of them. In this study, the expression levels of IFNγ mRNA in the spleen were almost the same between the extract- (PCPS-) treated group and control group. However, the expression of IL-4 mRNA showed a lower level in the extract-treated group than that in the control. Our results suggested that the PCPS could induce proinflammatory action in the immune response. In addition, the proinflammatory effect of the PCPS on THP-1 was enhanced by 5'-GMP-Na, while it was reduced by vitamin D2. These two compounds are majorly contained in the P. citrinopileatus mushroom. Therefore, these results suggested that the P. citrinopileatus mushroom might contain other immune regulative compounds, such as vitamin D2, as well as PCPS.

Characterization of the Lentinula edodes exg2 gene encoding a lentinan-degrading exo-beta-1,3-glucanase.

Lentinan, an antitumor substance purified from Lentinula edodes, is degraded during post-harvest preservation as a result of increased glucanase activity. We isolated an exo-beta-1,3-glucanase encoding gene, exg2, from L. edodes which is a homologue of an exo-glucanase-encoding gene conserved in ascomycetous fungi. The exg2 gene was cloned as an approximately 2.4-kbp cDNA, and as a genomic sequence of 3.9-kbp. The product of the exg2 gene is predicted to contain 759 amino acids with a molecular weight of 79 kDa and a pI value of 4.6. The putative N-terminus of EXG2 is identical to the N-terminal sequences of lentinan-degrading enzymes, GNase I and II, and a custom-made anti-EXG2 peptide anti-serum cross-reacted with purified GNase I and II. Transcription and translation of exg2 was low in the gills of mature fruiting bodies, but increased after harvesting. We conclude that the exg2 gene is a lentinan-degrading enzyme-encoding-gene in L. edodes.

Lemon flavonoid, eriocitrin, suppresses exercise-induced oxidative damage in rat liver.

To examine the preventive effect of the lemon flavonoid, eriocitrin (eriodictyol 7-O-rutinoside), on oxidative stress during acute exercise in vivo, levels of N( epsilon )- (hexanoyl)lysine, HEL; o,o-dityrosine, DT; and nitrotyrosine, NT, as oxidative stress markers, were determined by ELISA in livers of trained rats in addition to thiobarbituric acid-reactive substance (TBARS). Eriocitrin administration prior to exercise significantly suppressed the increases in TBARS caused by lipid peroxidation during acute exercise. The contents of HEL, DT, and NT in rat liver increased dramatically by exercise without eriocitrin administration. However, these increases were significantly suppressed by eriocitrin administration before exercise. Moreover, in this study, to clarify whether eriocitrin influences glutathione metabolite system that is considered to be important for a defense against the damage by oxidative stress, the levels of glutathione in rat liver were determined during exercise. The level of reduced glutathione after exercise was maintained by administration of eriocitrin. The increase in the concentration of oxidized glutathione caused by exercise was significantly suppressed by eriocitrin. This result suggested that eriocitrin might play an important role in the control of the change in glutathione redox status in rat liver during exercise. These findings showed that eriocitrin was effective in the prevention of oxidative damages caused by acute exercise-induced oxidative stress.