Application of lycium ruthenicum polysaccharides in preparation of immunoregulation medicines or health-care products
An immunomodulatory drug, the technology of Lycium barbarum, applied in application, food preparation, drug combination and other directions
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Embodiment 1
[0017] Example 1 Animal experiments to evaluate the immune activity of Lycium barbarum polysaccharide
[0018] (1) Tested sample: Lycium barbarum polysaccharide, prepared by the Glycotechnology Research Group of Dalian Institute of Chemical Physics, Chinese Academy of Sciences.
[0019] The preparation method of Lycium barbarum polysaccharide (LRP): add 4 times the volume of water to the fruit of Lycium barbarum and crush it to a uniform state with a tissue grinder, then heat and ultrasonic method (extraction conditions: temperature 70°C, ultrasonic frequency 40KHz, power 50W) Extract for 2 hours, centrifuge the extract (6000r / min, 10min), reheat the residue with 2 times the volume of water and ultrasonically extract for 1 hour, combine the two extracts, concentrate, ethanol precipitation with 4 times the volume of ethanol for 24 hours, filter with suction, and use for precipitation Dissolve 50 times the volume of water, add Sevage reagent [chloroform:n-butanol=4:1 (V / V)] to t...
Embodiment 2
[0050] Example 2 Effect of Lycium barbarum polysaccharides on splenocyte proliferation
[0051] The splenocyte suspension was prepared by conventional experimental methods, and polysaccharide samples with concentrations of 10, 50, 100, and 200 μg / mL were prepared; the splenocytes were prepared into a cell suspension containing 20% calf serum, and the concentration was 5×10 6 cells / mL; according to the normal control group, LPS stimulation group, and polysaccharide sample stimulation group, add cell suspension in 96-well culture plate in 3-4 replicate wells / group; Add 0.1mL / well of LPS or sample; 16h before terminating the culture, add 3 H-TdR 20 μl, so that the final concentration is 1 μci / mL; use a micro-manifold cell harvester to suck the cells onto 49-type glass fiber filter paper and wash them thoroughly with distilled water (washing 10 times); the glass fiber filter paper is dried overnight at room temperature; The next day, put the dried filter paper into a sealed bag...
Embodiment 3
[0053] Example 3 TNF-αELISA detection
[0054] Lycium barbarum polysaccharide (LRP) and its five main components (LRP1-LRP5) were respectively prepared into 10 mg / mL storage solution with PBS, and diluted with medium to a working concentration of 1 mg / mL for drug treatment in mice. Phage cells RAW264.7 were purchased from the Cell Bank of Shanghai Institute of Biological Sciences, Chinese Academy of Sciences, stored in liquid nitrogen, and cultured in DMEM medium containing 10% fetal bovine serum (Gibco). They were cultured in an incubator at 37°C with saturated humidity, 5% CO2 and 95% air, digested with 0.25% trypsin and passaged. RAW264.7 cells at 1×10 6 The density of cells / well was inoculated in a 24-well plate. After 24 hours of adherence, the culture medium was replaced with each group of drugs diluted with the culture medium. The drug concentration was 1mg / mL, and each group had two duplicate holes. The culture plate was placed in an incubator to continue culturing f...
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