Antimetastatic and Immunomodulating Effect of Water Extracts From Various Mushrooms

M8, β-glucan and pachyman were dissolved in 90% HPLC-grade methanol and applied to a prewashed silica gel 60 F254 HPTLC plate. M8 (2 or 4 mL of a 50 mg/mL solution) and 2 μL β-D-(1,3)-(1,6)-glucan (10 mg/mL) were separated (migration distance 60 mm) using HPLC-grade n-butanol/methanol/water (50:25:20). Images were visualized at UV 254 nm (A) or under white light after staining with aniline-diphenylamine-phosphoric acid (B).

(A) Female BALB/c mice (n = 6) were inoculated intravenously with colon26-L5 cells (2 × 10⁵ cells/mice). M8 prescription at the indicated doses was administered orally for 12 consecutive days from day 3 after tumor inoculation. 15 days after tumor inoculation, mice were sacrificed and lung weight was measured. (B) The body weight of all the living mice (n = 6) were measured and there was no statistical difference between the groups.

Female BALB/c mice were administered M8 orally for 2 weeks. One day after the last administration, mice were sacrificed and the splenocytes (1 × 10⁵ cells/well) suspended in RPMI-1640 medium supplemented with 10% FBS were cultured with or without Con A or LPS for 48 hours. XTT assay was conducted to evaluate cell proliferation. The data represent the mean ± S.D. *p < 0.05 as compared with control group.

Female BALB/c mice were orally administered M8 for 2 weeks. One day after the last administration, mice were sacrificed and the splenocytes (1 × 10⁵ cells/well) were stained with FITC- and PE-conjugated monoclonal anti- CD3, CD19, CD4, CD8, Mac-1 and NK1.1 antibodies. Stained cells were analyzed on flow cytometry using Cell Quest Software. A representative image of whole lymphocytes sorted using light scatter is shown.

Effect of oral administration of M8 on production of IFN-γ and IL-4 from splenocytes.