4-Hydroxy-17-methylincisterol from Agaricus blazei Decreased Cytokine Production and Cell Proliferation in Human Peripheral Blood Mononuclear Cells via Inhibition of NF-AT and NF-B Activation
Figure 4
Effects of 4-HM on cytokine gene expression in PBMC, as detected with RT-PCR. PBMCs () were cultured with PHA (5 μg/mL) or 2.5, 5, and 10 μM 4-HM for 18 hr. The total cellular RNA was isolated from PBMC and aliquots of RNA (1 μg) were reverse-transcribed for synthesis of cDNA. PCR was done as described in Section 2. (a) Following the reaction, the amplified products were taken out of the tubes and run on 2% agarose gel. Lane 1: medium, Lane 2: 0.1% DMSO, Lanes 3 to 5: 2.5, 5, or 10 μM 4-HM, Lane 6: PHA, Lane 7: PHA + 0.1% DMSO, Lanes 8 to 10: PHA + 2.5, 5, or 10 μM 4-HM. Each band was quantitated using laser scanning densitometer SLR-2D/1D (Biomed Instruments Inc., Fullerton, CA, USA). The ratios of (b) IL-2, (c) IL-4, (d) IFN-, or (e) TNF-mRNA to GAPDH mRNA were calculated. Each bar is the of three independent experiments with PBMC from different individuals. , : versus DMSO group.
