A kind of mulberry proteoglycan, its preparation method and application
A technology of proteoglycan and polysaccharide, applied in the field of mulberry proteoglycan, its preparation and application
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Embodiment 1
[0053] The preparation of embodiment 1 morula proteoglycan FMP-6-S4
[0054] a. Proteoglycan extraction:
[0055] 1 kg of dried mulberries was pulverized by a pulverizer, and 20 L of deionized water was added. At 55 °C, 30 g of cellulase, 10 g of amylase and 5 g of papain were added for extraction for 1 hour, and the temperature was raised to 100 °C to inactivate the enzyme, and centrifuged , the filtrate was concentrated and dialyzed against flowing water for 3 days. Heat and concentrate the dialyzed internal solution to 3L, centrifuge to discard the precipitate, add five times the volume (15L) of 95% v / v ethanol to the supernatant under stirring, let it stand for alcohol precipitation overnight, and centrifuge, and the obtained precipitate is washed with anhydrous Alternately washed with ethanol and acetone for 3 times, the precipitate was collected by centrifugation and vacuum-dried at 50° C. to obtain 152 g of crude proteoglycan from mulberry fruit (yield: 15.2%).
[005...
Embodiment 2
[0071] Example 2. Proteoglycan FMP-6-S4 inhibits Aβ 42 generation
[0072] 1. Aβ in CHO / APPBACE1 cells and HEK 293-APPsw cells 42 ELISA detection
[0073] CHO / APPBACE1 cells (from Shanghai Institute of Materia Medica, Chinese Academy of Sciences) were cultured in Ham's F12 medium (purchased from American Hyclone Company), HEK293-APP sw Cells (donated by Zhang Haiyan's research group at Shanghai Institute of Materia Medica, Chinese Academy of Sciences) were cultured in DMEM high-glucose medium containing 10% fetal bovine serum, 100 U / mL penicillin, 100 μg / mL streptomycin and 200 μg / mL G418. When the cells grow to 80%-90% confluence, 5×10 5 The density of cells / well was seeded in a 24-well plate in 5% CO 2 , After culturing in a 37°C incubator for 24 hours, add mulberry polysaccharide FMP configured at different concentrations (0 μg / mL, 31.25 μg / mL, 62.5 μg / mL, 125 μg / mL, 250 μg / mL, 500 μg / mL and 1000 μg / mL) -6-S4 (prepared in Example 1), after 24 hours, the cell supernata...
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