Application of agaricus gennadii gallic acid to functional food
A technology of gallic acid and torutospora, which is applied in the field of functional food, can solve the problem of blankness in the activity of chemical components, and achieve the effect of good esophageal cancer tumor, simple, stable and efficient process, and auxiliary inhibition of esophageal cancer tumor
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Embodiment 1
[0040] Preparation of the crude extract of each active component in the mushroom tortuosporum of embodiment 1
[0041] Take 10kg of fresh, undamaged Agaricus torusum fruiting body, slice it and soak it with 30L 95% ethanol for 7 days (wherein the ratio of Agaricus toricosporus fruiting body to ethanol is 1:3, so that the ethanol can completely soak the Agaricus toricosporin fruiting body), filter The filtrate was obtained, the solvent was recovered under reduced pressure, and 30 L of 95% ethanol was added to the filter residue for ultrasonic extraction 3 times, each time for 2 hours. Concentrate under reduced pressure to obtain 95% ethanol extract. Operate in the same way with 65% ethanol, and concentrate under reduced pressure to obtain 65% ethanol extract. Add 30L of distilled water to the filter residue of the mushroom after alcohol extraction, and extract three times with 1800HZ ultrasound, each time for 2 hours. Concentrate under reduced pressure to obtain 538 g of wate...
Embodiment 2
[0046] Add 60.0 g of the n-butanol fraction obtained in Example 1 above to ethanol, and crystals are precipitated on the container wall during precipitation. Repeat the operation and recrystallize multiple times to obtain 30 mg of compound 23. The n-butanol fraction was reversed phase C via medium pressure 18 Separation was performed by gradient elution with water-methanol solution (5% to 100%), the eluents were combined according to the same gradient, and the solvent was recovered under reduced pressure to obtain 6 components Z-I to Z-VI. The Z-Ⅱ component was separated by MCI resin, gradient eluted with water-methanol solution (0% to 100%), and washed with C 18 Semi-preparative column separation and purification yielded 4 mg of compound 7, 5 mg of compound 8, 4 mg of compound 9, and 5 mg of compound 10. The Z-IV fraction was separated and purified by a C18 semi-preparative column to obtain 5 mg of compound 6. The Z-I component is the pure water elution part of the MCI resi...
Embodiment 4
[0101] Example 4 Agaricus torusum gallic acid can inhibit the activity of luciferase with BRE reporter gene induced by BMP2
[0102] The C2C12 cell line was purchased from the Cell Bank of the Chinese Academy of Sciences, and the C2C12-pGF1-BRE cell line was purchased from the Shanghai Institute of Materia Medica, Chinese Academy of Sciences and stored in liquid nitrogen. C2C12-pGF1-BRE cells were cultured in DMEM medium containing 10% Gibco fetal bovine serum at 37°C with saturated humidity and 5% CO 2 cultured in an incubator at 1 x 10 4 The density of cells / well was seeded in a 96-well plate, 100 μL per well. After 24 hours of attachment to the wall, 60 μL of the medium was sucked off, and 50 μL of a sample prepared at a corresponding concentration (Gallic acid from Agaricus torusum in Example 1) was added, and the final concentrations were 1.5 mg / mL, 1 mg / mL, 0.5 mg / mL, and 10 μL, respectively. BMP2 was adjusted to a final concentration of 200ng / mL. At the same time, a ...
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