Metabolomic analysis of meju during fermentation by ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF MS).

Changes in the water-soluble metabolites of meju during fermentation were analysed by ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF MS), and the resultant data were statistically processed by partial least squares-discriminant analysis (PLS-DA). Various metabolites, including amino acids, small peptides, nucleosides, urea cycle intermediates, and organic acids, which are responsible for the unique taste and nutritional and functional quality of fermented soy foods, were clearly altered by increasing the fermentation period. Changes in these metabolites allowed discrimination among meju samples with different fermentation periods (0, 10, 20, 40, and 60d) on a PLS-DA score plot, and the fermentation was mainly processed between 10 and 40d of fermentation. Twenty-two metabolites (phenylalanine, glutamic acid, leucine, adenine, citrulline, arginine, glutamine, γ-aminobutyric acid, proline, acetylornithine, valine, pipecolic acid, methionine, citric acid, xanthine, tyrosine, isoleucine, Glu-Tyr, Ser-Pro, tryptophan, Glu-Phe, and Leu-Val-Pro-Pro) with high PLS-DA values of over 1.00 were determined as the major compounds contributing to the discrimination of meju samples. These metabolites, which were positively related to the sensory quality of meju, can be used as fermentation biomarkers for the production of meju and to construct the meju fermentation metabolic pathway. Therefore, our results indicate that monitoring the changes in metabolites during meju fermentation might be important for producing meju-related foods with good nutritional and sensory quality.

Cordyceps militaris extract suppresses dextran sodium sulfate-induced acute colitis in mice and production of inflammatory mediators from macrophages and mast cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Cordyceps militaris is a well-known medicinal mushroom used for treatment of asthma, and other bronchial and lung inflammatory diseases. AIM OF THE STUDY: To investigate the anti-inflammatory effects and mechanism of Cordyceps militaris extract on a murine model of acute colitis. MATERIALS AND METHODS: We induced colitis using DSS for 1 week. The disease activity index (DAI) took into account body weight loss, diarrhea, and bleeding. Colon length and crypt length were measured using a microscope. Structural changes of the colon were observed by H&E staining. NO, iNOS, and TNF-α were determined using the Griess assay. iNOS protein was determined using western blotting and quantitative reverse-transcriptase polymerase chain reaction, respectively. Degranulated mast cells in colon tissue were stained using toluidine blue. The degree of degranulated RBL-2H3 cells was measured by the ß-hexosaminidase assay. RESULTS: Cordyceps militaris extract significantly attenuated DSS-induced DAI scores (e.g., body weight loss, diarrhea, gross bleeding). Cordyceps militaris extract also effectively prevented shortening of colon length and crypt length. Histological analysis indicated that Cordyceps militaris extract suppressed epithelial damage, loss of goblet cells, loss of crypts, and infiltration of inflammatory cells induced by DSS. In addition, Cordyceps militaris extract inhibited iNOS and TNF-α mRNA expression in colon tissue of DSS-induced colitis and in LPS-stimulated RAW264.7 cells. Cordyceps militaris extract suppressed degranulation of mast cells in the colon of mice with DSS-induced colitis and in antigen-stimulated mast cells. CONCLUSION: These results suggest that Cordyceps militaris extract has anti-inflammatory activity in DSS-induced acute colitis by down-regulating production and expression of inflammatory mediators. These findings suggest that Cordyceps militaris extract might be applied as an agent for prevention or treatment of inflammatory bowel diseases (IBDs).

The ethyl acetate extract of PGP (Phellinus linteus grown on Panax ginseng) suppresses B16F10 melanoma cell proliferation through inducing cellular differentiation and apoptosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Phellinus linteus and Panax ginseng have been widely used as traditional herbal medicines to treat various diseases including cancer in East Asia. AIM OF THE STUDY: The present study sought to investigate the possible mechanism in anti-proliferative effect of Phellinus linteus that was grown on Panax ginseng (PGP) on B16F10 melanoma cells. MATERIALS AND METHODS: The anti-proliferative effect of PGP on B16F10 was evaluated by CCK-8 assays. Apoptotic cells were detected by flow cytometry analysis. The proteins involved in apoptosis and cellular differentiation were assessed by immunoblot analysis. Ginsenosides contents of PG or PGP were analyzed using HPLC. RESULTS: The ethyl acetate fraction (EtOAc) of PGP exhibited the strongest anti-proliferative activity among PGP fractions (butanol or water) on B16F10 cells. PGP EtOAc extract showed stronger inhibitory effect than Panax ginseng (PG) or Phellinus linteus (PL) EtOAc extract on B16F10 melanoma cell proliferation. PGP EtOAc extract induced the dendrite-like structures and the melanin production in B16F10 cells. PGP EtOAc extract increased a sub-G1 cell population through inducing p53/p21 and activated caspase-8 protein expression in B16F10 cells. Notably, PGP EtOAc extract contained ginsenosides Rd, Rg3, Rb2, Rg1 and Rb1 more than PG EtOAc extract. Rd and Rg3 significantly inhibited B16F10 cell proliferation. CONCLUSION: Our data suggest that PGP EtOAc extract inhibits B16F10 cell proliferation through inducing apoptosis and cellular differentiation.

An extract of Phellinus linteus grown on germinated brown rice inhibits inflammation markers in RAW264.7 macrophages by suppressing inflammatory cytokines, chemokines, and mediators and up-regulating antioxidant activity.

The immunomodulatory activity of an organic extract of Phellinus linteus grown on slightly germinated brown rice (PBR) was previously demonstrated. Here, we investigated the possible anti-inflammatory activity of the PBR extract by analyzing its effect on the expression of macrophage-derived cytokines, chemokines, and mediator genes that participate in immune and inflammatory responses and diseases. The extract profoundly inhibited the induction of cytokines and chemokines, including tumor necrosis factor-α, chemokine (C-X-C motif) ligand-10, granulocyte-macrophage colony-stimulating factor, and interleukin-6, in lipopolysaccharide (LPS)-stimulated RAW264.7 mouse macrophage cells. It also greatly inhibited LPS-stimulated production of nitric oxide (NO) and prostaglandin E(2) in RAW264.7 cells by suppressing the expression of inducible NO synthase and cyclooxygenase-2. PBR extract inhibited NO production with a twofold lower half-maximal inhibitory concentration value than P. linteus extract. To elucidate the underlying mechanism of action, we examined the effect of the PBR extract on the LPS-induced phosphorylation of mitogen-activated protein kinases (MAPKs) in RAW264.7 cells. PBR extract greatly inhibited extracellular signal-regulated kinase and c-Jun N-terminal kinase phosphorylation and slightly inhibited p38 MAPK phosphorylation. It also significantly increased intracellular glutathione peroxidase activity and heme oxygenase-1 protein expression. Thus, the PBR extract has anti-inflammatory activity in LPS-stimulated RAW264.7 cells by virtue of its ability to suppress the production of inflammatory cytokines and chemokines via inhibition of MAPK activation and up-regulation of antioxidant activities.