A bright moon grass polysaccharide and its application in the preparation of drugs and functional foods for immunoregulation and anti-tumor
A technology of functional food and immune regulation, applied in the field of medicine, can solve problems such as no related research, and achieve the effect of inhibiting tumor growth, increasing the level of white blood cells, and inhibiting migration
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Embodiment 1
[0022] Example 1, Extraction, preparation and analysis of Mingyuecao polysaccharide
[0023] The preparation method of Mingyuechao polysaccharide is as follows: take 50g of Mingyuechao stem and leaf powder, add 20 times the mass of deionized water, reflux extraction at 80-100°C for 2 hours, centrifuge, take the supernatant, and add 5 times the mass of deionized water for precipitation, Reflux extraction at 80-100°C for 2 hours, centrifuge, combine the filtrate, add a certain amount of 95% ethanol to the filtrate to make the final concentration of ethanol 75%, ethanol precipitation overnight, centrifuge to remove the supernatant, the precipitate is deproteinized and decolorized, and freeze-dried. The finally obtained biologically active polysaccharide is the moon grass polysaccharide of the present invention.
[0024] According to the HPLC molecular weight determination of the described Crescent Moon grass polysaccharide, the weight average molecular weight is 20-2000KDa, the t...
Embodiment 2
[0025] Example 2, the promoting effect of Mingyuecao polysaccharide on the phagocytosis of neutral red by RAW264.7 cells
[0026] Mouse mononuclear macrophage RAW264.7 cells were cultured in DMEM high glucose medium containing 10% fetal bovine serum. Cells in the logarithmic growth phase were taken at 1.0×10 5 The concentration of cells / mL was inoculated in a 96-well plate in 5% CO 2 , 37°C, cultured in a constant temperature incubator with saturated humidity until the cells adhered to the wall (about 6-8h), and then added 6 concentrations of polysaccharides containing different concentrations (1,5,10,25,50,100μg / ml) , the positive drug is lentinan, the final concentration is 2 μg / ml, set 4 parallel wells for each concentration, take it out after 12 hours of culture, absorb the original medium, wash it once with PBS at room temperature, add 100 μL 0.075% neutral red dye to each well solution, continue to incubate for 30min, absorb the neutral red, wash twice with PBS, lyse w...
Embodiment 3
[0032] Example 3, the effect of Mingyuecao polysaccharide on promoting the secretion of nitric oxide from RAW264.7 cells
[0033] Mouse mononuclear macrophage RAW264.7 cells were cultured in DMEM high glucose medium containing 10% fetal bovine serum. Cells in the logarithmic growth phase were inoculated in 96-well plates at a concentration of 1.0x105 cells / mL, cultured in a constant temperature incubator with 5% CO2, 37°C, and saturated humidity until the cells adhered to the wall (about 6-8 hours), and then added There are 6 concentrations of polysaccharides of different concentrations (1, 5, 10, 25, 50, 100 μg / ml), the positive drug is lentinan, the final concentration is 2 μg / ml, 4 parallel wells are set for each concentration, and after 24 hours of incubation Take it out, take 50 μL of the culture medium into another 96-well plate, and detect it according to the instruction of the nitric oxide detection kit (Beyontian Bioreagent Co., Ltd.), that is, the Grignard reagent me...
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