Comparison of the effects of various species of mushroom on the fatty acid synthesis pathway.

<p>(A) <i>FAS</i> and (B) <i>ELOVL6</i> gene expressions in HepG2 cells treated with various species of mushroom extract (WBM, shiitake mushroom, enoki mushroom and oyster mushroom) for 24 hours. Real time PCR analysis was performed for the gene, <i>ELOVL6 and FAS</i>. Gene expression was normalized with the <i>β-actin</i> housekeeping gene. Values are expressed as mean and standard error. We analyzed the data for each dose separately by ANOVA, followed by Tukey's multiple comparison test. Statistical significance was defined as <i>P</i><0.05 compared to control.</p

Effects of WBM feeding on body weight, liver weight, ALT serum levels and uterus weight.

<p>Seven-week old female C57B1/6J mice were ovariectomized. These mice are a model of postmenopausal women (OVX, n = 16). Control mice (sham, n = 16) were subjected to sham operation survival surgery. Both sham and OVX mice were fed a HFD (n = 8/per group) or HFD with WBM (n = 8/per group) diet for 3 months. Body weight was measured once a week. (A) Body weight measurement of sham mice. (B) Body weight measurement of OVX mice. (C) Liver weight. (D) ALT serum levels. (E) Uterus weight. Values are expressed as mean and standard error for 8 mice. * Statistical significance was defined as <i>P</i><0.05.</p

LXR and ER activity in HepG2 cells treated with WBM extract.

<p>(A) HepG2 cells were transfected with the pCMX-VP16-hLXRa and LXR-responsive rCYP7A-DR-4x3-tk-LUC, and following were incubated with vehicle control (ethanol), T0901317 (10 µM) and/or WBM extract (1, 2 and 5 µl/ml) and assayed for luciferase activity. (B) For evaluation of WBM effects on ER activity, HepG2 cells were transiently transfected with the pSG5-ER and pGL₃(ERE)₃-Luc. 24 hours posttransfection, cells were treated with E2 (0.1 nM) or with WBM extract (5 and 10 µl/ml) for 24 hours. Data is expressed as relative luciferase unit/protein content. Values are expressed as mean and standard error. For LXR activity, we analyzed the data for each dose separately by ANOVA, followed by Tukey's multiple comparison test. * <i>P</i><0.05 compared indicated treatment, a; <i>P</i><0.05 compared to T0901314 treatment. For ER activity, we analyzed the data for each treatment by ANOVA, followed by comparison of all treatment groups with the control group (Dunnett's test). Statistical significance was defined as <i>P</i><0.05.</p

WBM intake decreased fat accumulation in the liver and improved glucose clearance ability in OVX mice.

<p>(A) Macroscopic appearance of liver from mice fed the HFD and HFD+WBM for 3 months in each sham and OVX group. The tissues were fixed with 10% formalin overnight and then embedded in paraffin. Five-micrometer sections were cut and stained with hematoxylin and eosin, and examined by light microscope. Representative H&E staining of livers from each group are shown. (B) Serum glucose concentration after glucose injection in mice fed with HFD or HFD+WBM diet for 2 months. The mice were challenged with 1.5 mg glucose/g body weight glucose load. The glucose levels pre, 30, 60, 120 and 180 min post injection were measured using a glucometer. Values are expressed as mean and standard error for 8 mice. * Statistical significance was defined as <i>P</i><0.05.</p

Composition of experimental diets.

<p>Powder was mixed to a modified AIN-93G diet enriched in fat (HF).</p>a<p>Carbohydrate: 43.30%, fiber: 13.20% protein: 28.80%, fat: 4.50%</p

Effects of methanol fractions from WBM extract on <i>FAS</i> and <i>ELOVL6</i> gene expressions and aromatase activity in HepG2 cells.

<p>Cells were incubated with each methanol fraction from WBM extract for 24 hours. (A) <i>FAS</i> and <i>ELOVL6</i> gene expression was normalized with the β-actin housekeeping gene. (B) Microsome assays were performed to evaluate anti-aromatase effects of WBM using the substrate, [1-β-³H] androstenedione. Activity was calculated to measure supernatant containing [³H] H₂O as reaction product, and then was counted in a Scintillation Counter. Aromatase inhibition activity was calculated as the percentage of remaining activity from the reaction without mushroom fractions. Analyses were carried out in triplicate and data were expressed as the mean ± SE. We analyzed the data for each treatment by ANOVA, followed by comparison of all treatment groups with the control group (Dunnett's test). Statistical significance was defined as <i>P</i><0.05.</p