Figure 1.
HPLC chromatogram of native β-D-glucan NG and three β-D-glucan phosphates, GP-2, GP-4, and GP-5 prepared by planetary ball milling.
Table 1.
β-D-glucan phosphates prepared under different operation conditions.
Figure 2.
FTIR spectra of insoluble β-D-glucan particles NG and soluble β-D-glucan phosphate GP-2 and GP-4 prepared by planetary ball milling.
Figure 3.
13C NMR spectra of insoluble β-D-glucan particles NG and β-D-glucan phosphate GP-2 prepared by planetary ball milling.
Figure 4.
31P NMR spectra of insoluble β-D-glucan particles NG, β-D-glucan phosphate GP-2 prepared by planetary ball milling, and sodium hexametaphosphate (NaPO3)6 milled alone.
Figure 5.
Effects of native β-D-glucan and three β-D-glucan phosphates, GP-2, GP-4, and GP-5, on cell proliferation (A) and neutral red uptake (B) of RAW264.7 cells. RAW264.7 cells were treated with NG or GP (50, 100, and 500 µg/mL) or LPS (10, 100, and 1000 ng/mL) in different concentrations as described in the Materials and Methods.
After incubation, the viability of RAW264.7 cells was measured by an MTT assay, and the A570 value was recorded, whereas the amount of neutral red uptake was detected by the A540 value. The data represent the means ± SD. *p<0.05, **p<0.01 compared with control. Each point represents the average of three independent experiments.
Figure 6.
Effects of native β-D-glucan and three β-D-glucan phosphates, GP-2, GP-4, and GP-5, on TNF-α (A) and IL-6 (B) production by RAW264.7 cells.
RAW264.7 cells were treated as Fig. 5. After incubation, the TNF-α and IL-6 concentrations in the supernatant were detected using commercial kits. The data represent the means ± SD. *p<0.05, **p<0.01 compared with control. Each point represents the average of three independent experiments.
Figure 7.
Effects of native β-D-glucan and three β-D-glucan phosphates, GP-2, GP-4, and GP-5, on NO secretion by RAW264.7 cells.
RAW264.7 cells were treated as Fig. 5. After incubation, the NO concentrations in the supernatant were detected by the A540 value after reacting with Griess reagent. The data represent the means ± SD. *p<0.05, **p<0.01 compared with control. Each point represents the average of three independent experiments.