Enteromorpha polysaccharide having immunocompetence and preparation method thereof
The technology of prolifera polysaccharide and immune activity is applied in the field of prolifera polysaccharide with immune activity and its preparation, which can solve the problems of wasting high-quality resources, affecting the appearance of water bodies, increasing treatment costs and the like, and achieving the effect of good immune activity.
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Embodiment 1
[0023] Embodiment 1: the preparation method of HT-II polysaccharide:
[0024] 1. Extraction: Take a sample of Enteromorpha from the sea area of Qingdao, wash the sample of Enteromorpha with tap water, take 100g after natural air-drying, break it up, add distilled water with the mass ratio of Enteromorpha slag and water as 1:30, and extract at 95°C After 2 hours, filter with suction, and extract the filter residue twice with 1:15 distilled water. The filtrates were combined, concentrated by rotary evaporation until the liquid was slightly viscous, 3 times the volume of 95% ethanol was added, and allowed to stand overnight at 4°C. Centrifuge at 9000rpm, dehydrate the precipitate with 85%, 95%, absolute ethanol and ether in turn, and freeze-dry to obtain the crude polysaccharide HT.
[0025] 2. Deproteinization: make the crude polysaccharide into 5% sugar solution, use papain, take protease into the sugar solution according to the mass ratio of enzyme and crude polysaccharide ...
Embodiment 2
[0027] Embodiment 2: the immune activity test of HT-II polysaccharide:
[0028] 1. The effect of HT-II polysaccharide on the proliferation of spleen lymphocytes: using MTS-PMS colorimetric method. BALB / c mice were sacrificed by breaking the cervical spine, and the spleen was taken aseptically to prepare splenic lymphocytes, and the cell concentration was adjusted to 410 with RPMI1640 complete culture medium 9 / L cell / ml.
[0029] BALB / c mice 410 9 / L cell / ml splenocyte suspension was added to a 96-well cell culture plate, 100 μl per well, and HT diluted with RPMI1640 complete culture medium was added to each well of the polysaccharide group to a concentration of: 6.25, 12.5, 25, 50, 100 mg / L 10 μl each of -II, 5 repetitions for each concentration, and the blank control group was replaced by an equal volume of RPMI1640. Set at 37°C, 5% CO 2 Cultivate for 72 hours in a saturated humidity incubator. Take it out, add 20μl MTS-PMS to continue culturing for 4-6 hours, and measu...
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