Physicochemical characteristics and anti-inflammatory activities of antrodan, a novel glycoprotein isolated from Antrodia cinnamomea mycelia
Figures
Figure 1
Gel filtration chromatogram of polysaccharides…
Figure 1
Gel filtration chromatogram of polysaccharides AC-2 prepared from AC mycelia using a Sepharose…
Figure 1 Gel filtration chromatogram of polysaccharides AC-2 prepared from AC mycelia using a Sepharose CL-6B column (3.0 × 82 cm). Fractions (5.0 mL/fraction) were collected and assayed for the contents of sugars (at 490 nm) and proteins (at 280 nm). The vertical dashed lines indicated the fractions collected for preparation of antrodan.
Figure 2
High performance size exclusion chromatograms…
Figure 2
High performance size exclusion chromatograms of antrodan detected at UV 280 nm and…
Figure 2 High performance size exclusion chromatograms of antrodan detected at UV 280 nm and ELSD absorption and the molecular weight estimation. The regression equation was obtained from the retention time vs. The MWs of pullunan reference set: 788, 404, 212, 112, 47.3, 22.8, 11.8, and 5.9 kDa. log Da = −0.466X + 10.009, R2 = 0.9937, where X is the retention time of the target polymer. Samples: antrodan, biobran, and yeast β-glucan.
Figure 3
Total ion chromatograms of GC/MS…
Figure 3
Total ion chromatograms of GC/MS analysis of monosaccharides. ( a ) authentic monosaccharides…
Figure 3 Total ion chromatograms of GC/MS analysis of monosaccharides. (a) authentic monosaccharides and (b) monosaccharides obtained from the hydrolyzed antrodan. Ara: arabinose, Rha: rhamnose, Fuc: fucose, Xyl: xylose, I.S.: internal standard arabitol, Man: mannose, Gal: galactose, Glc: glucose. *: Mannitol.
Figure 4
Comparison of the FTIR spectra.…
Figure 4
Comparison of the FTIR spectra. ( a ) antrodan, ( b ) biobran,…
Figure 4 Comparison of the FTIR spectra. (a) antrodan, (b) biobran, and (c) yeast β-glucan. 2 mg of pure dry sample and 300 mg of pure dry KBr were mixed and pressed into a disc. The whole IR spectrum (400 to 4000 cm−1) was recorded on a FTIR spectrophotometer. Biobran is a reference of pure polysaccharide and yeast β-glucan acts as the β-glucan reference.
Figure 5
( a ) 1 H-NMR…
Figure 5
( a ) 1 H-NMR spectra of antrodan and ( b ) a…
Figure 5 (a) 1H-NMR spectra of antrodan and (b) a standard β-d-(1→3, 1→6)-linked glucan of curdlan.
Figure 6
Effect of ( a )…
Figure 6
Effect of ( a ) antrodan and biobran, and ( b ) LPS…
Figure 6 Effect of (a) antrodan and biobran, and (b) LPS on the cell viability of RAW 264.7 mouse macrophages. Cells were incubated with antrodan or biobran for 24 h (a) and incubated with lipopolysaccharide (LPS; 0.05~5 μg/mL) for 24, 48 and 72 h, respectively. Cell proliferation was evaluated by the MTT assay. Values are expressed as mean ± SEM of three independent experiments. * p < 0.05 vs. Control; ** p < 0.01; *** p < 0.001.
Figure 7
Effect of LPS on (…
Figure 7
Effect of LPS on ( a ) NO production in RAW 264.7 mouse…
Figure 7 Effect of LPS on (a) NO production in RAW 264.7 mouse macrophages, (b) 24 h treatment, (c) 48 h treatment and (d) 72 h treatment with antrodan and biobran. Cells were treated with various concentrations of LPS for 24, 48 and 72 h, respectively. Nitrite content in the cultured medium was determined by a Griess reaction assay. Values are expressed as mean ± SEM of three independent experiments. The confidence levels * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. control were determined by the Student t-test. All figures (7)