Bioguided isolation, identification and bioactivity evaluation of anti-fatigue constituents from Schizophyllum commune.

This study aims to clarify the specific anti-fatigue components of Schizophyllum commune (S.commune) and analyze its potential anti-fatigue mechanism. The main anti-fatigue active ingredient of S.commune was locked in n-butanol extract (SPE-n) by activity evaluation. Twelve compounds were identified by high performance liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS). The anti-fatigue effect of morusin is the most predominant among these 12 ingredients. The determination of biochemical indices showed that morusin could increase liver glycogen reserves, improve the activity of antioxidant enzymes in liver, and reduce reactive oxygen species (ROS) content in muscle tissue, thereby reducing myocyte damage. Further studies revealed that morusin could reduce the level of oxidative stress by activating Nrf2/HO-1 pathway, thus alleviating the fatigue of mice caused by exhaustive exercise. The current findings provide a theoretical basis for the development of natural anti-fatigue functional food.

Improved production of polysaccharides in Ganoderma lingzhi mycelia by plasma mutagenesis and rapid screening of mutated strains through infrared spectroscopy.

As a traditional Chinese medicine, Ganoderma lingzhi has attracted increasing attention for both scientific research and medical application. In this work, in order to improve the production of polysaccharides from an original wide-type (WT) strain (named "RWY-0") of Ganoderma lingzhi, we applied atmospheric-pressure dielectric barrier discharge (DBD) nonthermal plasma to the protoplasts of RWY-0 for mutagenesis treatment. Through a randomly amplified polymorphic DNA (RAPD) assay, at least 10 mutagenic strains were confirmed. They also showed different mycelium characteristics in terms of shape, color, size and biomass in liquid fermentation. The mutant strains were examined by infrared spectroscopy, and based on the established near-infrared (NIR) quantification model, the polysaccharide contents in these mutants were quantitatively evaluated. As a result, we found that the Ganoderma polysaccharide contents in some of the mutant strains were significantly changed compared with that of the original WT strain. The polysaccharide content of RWY-1 G. lingzhi was considerably higher than that of the WT strain, with an increase of 25.6%. Thus, this preliminary work demonstrates the extension of the plasma mutagenesis application in acquiring polysaccharide-enhanced Ganoderma lingzhi mutants and shows the usefulness of NIR spectroscopy in the rapid screening of mutagenic strains for other important ingredients.

[Effect of pulchinenoside on FZD8 expression of adjuvant arthritis rats].

To study the effect of pulchinenoside (PULC) on the Frizzled (FZD) expression of adjuvant arthritis ( AA) rats. AA rats were prepared through the toe injection with complete Freund's adjuvant to culture fibroblast-like synoviocytes (FLS). The effect of the oral administration with PULC on the FZD8 expression was detected by the real time qPCR. The effect of FZD8 knockout on the expressions of IL-1, IL-6, IL-8 were detected by MTT and ELISA. The role of miR-375 in the abnomal expression of FZD8 was detected by the real time qPCR. The results showed signfiicant decrease in the FZD8 expression among AA rats, FLS proliferation ater FZD8 knockout and IL-1, IL-6, IL-8 expressions and notable increase in miR-375 expression after the oral administration with PULC. The up-regulated miR-375 expression can inhibit the FZD8 expression. PULC may inhibit the FZD8 expression by up-regulating the miR-375 expression.

[Pulchinenoside control MeCP2 expression in FLS from RA model rats].

The role of pulchinenoside (PULC) in the regulation of MeCP2 expression was investigated in RA model rats. Adjuvant arthritis rats were used as RA model rats, and fibroblast-like synoviocytes (FLS) from the RA model rats were cultured. The effect of 100 mg x kg(-1) PULC gavage treatment on the MeCP2 expression and the effect of MeCP2 siRNA on the expression of SFRP2 and ß-catenin were detected by real time qPCR and Western blotting. The role of PULC in the FLS proliferation was detected by MTT. The results showed that the MeCP2 expression was down-regulated, the SFRP2 expression was up-regulated and the FLS proliferation was inhibited in FLS after therapy. MeCP2 siRNA significantly inhibited the MeCP2 expression, up-regulated the SFRP2 expression and inhibited the ß-catenin expression in FLS from RA model rats. PULC may increase the SFRP2 expression, inhibit the Wnt signaling and inhibit the FLS proliferation in FLS from the RA model rats by inhibiting the MeCP2 expression.

[Effect of pulchinenoside in regulating FLS SFRP2 expression of RA model rats].

OBJECTIVE: To study the effect of pulchinenoside (PULC) in modulating SFRP2 expression in fibroblast-like synoviocytes (FLS) of rheumatoid arthritis (RA) model rats. METHOD: The effect of PULC in treating RA rats was evaluated by rat arthritis score and paw swelling score. The inhibitory effect of PULC on FLS proliferation was detected by MTT reagent. The effects of PULC gavage treatment in modulating gene expression of FLS SFRP2, critical gene beta-catenin of Wnt pathway and downstream effector genes C-myc of of Wnt pathway were detected by RT-PCR and Western blotting. RESULT: PULC had a significant effect in treating RA rats and that SFRP2 expression was down-regulated in FLS. After PULC gavage treatment, FLS SFRP2 expression was obviously up-regulated, whereas beta-catenin and C-myc gene expressions were significantly down-regulated. CONCLUSION: PULC can inhibit abnormal proliferation of synovial membrane by modulating Wnt pathway of RA rats.

[Chuju total flavonoids control the SFRP4 expression in Wnt pathway in rheumatoid arthritis model rats].

OBJECTIVE: To determine the effect of Chuju total flavonoids (CJTF) on the secreted frizzledrelated protein 4 (SFRP4) expression in Wnt pathway in rheumatoid arthritis (RA) model rats. METHODS: The role of CJTF in the treatment of RA model rats was evaluated by rat arthritis score and paw edema score. The expression regulation of the SFRP4, ß-catenin and C-myc in Wnt pathway in RA model rats was detected by RT-PCR and Western blot after CJTF gavage treatment. RESULTS: After CJTF treatment, the rat arthritis score and paw edema score in RA model rats were significantly decreased when the RA model rats were treated with CJTF, the SFRP4 expression was significantly up-regulated, while the ß-catenin and C-myc gene expression were significantly downregulated in RA model rat synovial tissues. CONCLUSION: CJTF has significant therapeutic effect and inhibitory effect on Wnt pathway activation by targeting SFRP4 in RA model rat synovium.