Traditional Chinese medicine compound polysaccharide with function of synergistically enhancing humoral and cellular immunity and application thereof
A compound polysaccharide and cellular immunity technology, which is applied in the fields of animal pharmacy, medicine and veterinary medicine, can solve the problems of mixed infection of bacterial diseases, destruction of the immune system of pigs, failure of vaccine immunity, etc., to increase the percentage of peripheral blood lymphocytes, strengthen spleen Lymphocyte proliferative ability and effect of enhancing ability
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[0025]1) Take 2Kg of fresh shiitake mushrooms and wash them with water, add 20Kg of distilled water to decoct for 1 hour according to the mass ratio, take the primary filtrate of shiitake mushrooms and collect the medicinal residues;
[0026] 2) Take 0.5Kg Astragalus Chinese herbal decoction pieces, add 5Kg distilled water according to the mass ratio and ultrasonically impregnate for 20 minutes, decoct for 1 hour, take the primary filtrate of shiitake mushrooms and collect the medicinal residues;
[0027] 3) Take 0.5Kg Codonopsis pilosula Chinese herbal decoction pieces, add 5Kg distilled water according to the mass ratio and ultrasonically impregnate for 20 minutes, decoct for 1 hour, take the primary filtrate of Codonopsis pilosula and collect the dregs;
[0028] 4) The medicinal residues in steps 1), 2), and 3) were added to 5 times the amount of distilled water according to the mass ratio and decocted for 30 minutes to obtain the secondary filtrates of mushrooms, astragalus...
experiment example 1
[0033] Experimental example 1: CCK8 method was used to detect the effect of polysaccharides of single traditional Chinese medicine and polysaccharides of compound traditional Chinese medicine on the proliferation of mouse spleen lymphocytes in vitro
[0034] Test method: Aseptically take male Balb / c mouse spleen with an age of 6-8 weeks and a body weight of 20g±2g, and blow out the cells in RPMI-1640 culture medium. Collect the cell suspension and use Tris-NH4Cl to remove red blood cells, resuspend the cell pellet with RPMI-1640 culture medium, count more than 95% of viable cells by Trypan Blue staining, and adjust the cell density to 5×10 6 cells / ml to make a spleen cell suspension. The grouping and drug administration are shown in Table 1. After the above cells and drugs were added and marked, the 96-well cell culture plate was placed at 37°C and 5% CO 2 Continuously culture in the incubator for 44 hours; take out the culture plate, add 20 μl CCK8 solution to each well, an...
experiment example 2
[0045] Experimental Example 2: Using Orthogonal Experiments to Screen the Optimal Ratio of Chinese Medicine Combination Stimulating Spleen Lymphocyte Proliferation in Mice
[0046] Aseptically take the spleen of healthy 6-8 week old Balb / c male mice and blow out the cells in RPMI-1640 culture medium. Collect the cell suspension and use Tris-NH4Cl to remove red blood cells, resuspend the cell pellet with RPMI-1640 culture medium, count more than 95% of viable cells by Trypan Blue staining, and adjust the cell density to 5×10 6 cells / ml to make a spleen cell suspension. According to the results in Table 2, the optimal concentrations of lentinan, astragalus polysaccharide, and Codonopsis polysaccharide to stimulate the proliferation of mouse spleen lymphocytes in vitro are 25 mg / L, 25 mg / L, and 100 mg / L, respectively, adding 100 μl; using orthogonal experiments to design polysaccharides Combine and add cells, add cells and drugs, after marking, place the 96-well cell culture pla...
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