Daun negokilo polysaccharide and application thereof in preparation of immunoregulatory and antineoplastic drug and functional food
A functional food and immune regulation technology, applied in the field of medicine, can solve problems such as no related research, and achieve the effects of inhibiting tumor growth, strong anti-angiogenesis, and inhibiting weight loss
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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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