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 5
4-HM did not affect ERK phosphorylation and c-fos gene expression in PBMC induced by PHA. (a) PBMCs ( cells) were treated with DMSO (0.1%), or 4-HM (2.5, 5, 10 μM) in the presence or absence of PHA (5 μg/mL) for 2 hr. Lysates (50 μg/mL) were collected and run on a 10% SDS-PAGE gel and analyzed by immunoblotting with anti-pERK, or ERK Ab. (b) Bar graph indicates the ratio of pERK to ERK proteins. (c) 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 RT-PCR was done as described in Section 2. Following the reaction, the amplified product was run on 2% agarose gel. (d) The ratio of c-fos mRNA to GAPDH mRNA was calculated. Each band was quantitated using laser scanning densitometer SLR-2D/1D (Biomed Instruments Inc., Fullerton, CA, USA). Each bar represents from three independent experiments with PBMC from different individuals. Lane 1: 0.1% DMSO, Lanes 2 to 4: 2.5, 5, or 10 μM 4-HM, Lane 5: % DMSO, Lanes 6 to 8: , 5, or 10 μM 4-HM.
