Nostoc commune antitumor polysaccharide as well as preparation method and application thereof
An anti-tumor and ground fungus technology, which is applied in the fields of anti-tumor drugs, applications, food preparation, etc., can solve the problems that the anti-tumor activity of ground fungus polysaccharides has not been seen, and achieve the inhibition of cancer cell proliferation, great development prospects and application value, The effect of good anticancer activity
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Embodiment 1
[0021] The preparation method of embodiment 1 fungus anti-tumor polysaccharide (NVPS)
[0022] (1) Dry the natural fungus at 60° C. for 2 hours, and crush it.
[0023] (2) Take 100 g of ground fungus powder, add 5 L of distilled water, heat and reflux for 3 hours, let stand, filter with double-layer gauze, take the supernatant, repeat the extraction once as above, combine the two filtrates, evaporate and concentrate to 1000 ml under reduced pressure;
[0024] (3) add 100ml potassium ferrocyanide (K 4 [Fe(CN) 6 ]·3H 2 O) (10.6%), shake well, then add 100ml zinc acetate (C 4 h 8 o 4 Zn) (21.9%), shake evenly to precipitate protein, place at room temperature for 30 minutes, centrifuge at 5000rpm for 20 minutes, take supernatant, evaporate and concentrate to 100ml under reduced pressure;
[0025] (4) The concentrated solution obtained in step (3) is dialyzed in distilled water with a dialysis bag with a cutoff of 3000, and the water is changed once every 12 hours until the c...
Embodiment 2
[0029] Example 2: Select MCF-7 cells in the logarithmic growth phase, make the cells into a cell suspension, count the cells, 4×10 cells per well 4 Inoculated into a 96-well plate at 37°C, 5% CO 2 Culture in the incubator for 24 hours and after the cells adhere to the wall, discard the old medium, add medium with different polysaccharide final concentrations (0, 25, 50, 100, 200, 300ug / ml), set 5 parallel wells, and culture The plate was further incubated for 24 hours, and 20 μL of 5.0 mg / mL MTT was added to each well, and incubated for another 4 hours. Remove the culture medium in the plate, add 150 μL of DMSO to each well to dissolve the formazan crystals produced by the living cells, shake on a shaker for 10 min, and measure the absorbance of each well at a wavelength of 570 nm with a microplate reader. A concentration was considered toxic to cells if the decrease in absorbance of wells containing polysaccharide extract was greater than 10% compared to control wells (ie, i...
Embodiment 3
[0032] Select MCF-7 cells in the logarithmic growth phase, adjust the cell concentration to 8×10 per well 3 cells / 100 μl of culture medium and added to a 96-well culture plate. 37°C, 5% CO 2 Incubate overnight in an incubator. After the cells adhered to the wall, the culture plate was taken out, and the culture medium containing different concentrations (0, 25, 50, 100, 200, 300ug / ml) of Auricularia polysaccharide was replaced, and 5 parallel wells were set. The culture plate was put back into the incubator to continue culturing for 24 hours and 36 hours, and 20 μL of 5.0 mg / mL MTT was added to each well, and the incubation was continued for 4 hours. Aspirate the culture medium in the plate, add 150 μl of DMSO to each well to dissolve the formazan crystals produced by the living cells, shake on a shaker for 10 min, and measure the absorbance of each well with a microplate reader at a wavelength of 570 nm.
[0033] The fungus polysaccharide (NVPS) obtained in the present inv...
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