Immunity inhibitor glycyrol
An immunosuppressive and immunosuppressive technology, which is applied in the directions of medical preparations containing active ingredients, plant raw materials, organic chemistry, etc. Application and other issues, to achieve the effect of easy modification, small molecular weight and simple structure
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Embodiment 1
[0014] Embodiment 1: the purification of glycyrrhizin
[0015] The dried licorice is crushed to a particle size of 10-20 mesh, put into a percolation device, extracted by ethanol percolation, and the percolation liquid is collected. The percolation liquid was concentrated under reduced pressure and then extracted with ethyl acetate. After the extract was concentrated, the sample was prepared, and then silica gel column chromatography was carried out, and the gradient elution was carried out with petroleum ether-ethyl acetate system, and the fractions were collected, and the solid precipitated fraction was retained . These fractions were purified by high-speed countercurrent chromatography (HSCCC) to obtain white needle-like crystals, and the hydrogen spectrum, carbon spectrum, hydrogen-hydrogen correlation spectrum, carbon-hydrogen correlation spectrum and mass spectrum of the nuclear magnetic resonance spectrum (NMR) The compound was analyzed as glycyrrhizol.
Embodiment 2
[0016] Example 2: Inhibitory effect of Glycyrrhizol on ConA-induced proliferation of mouse T lymphocytes
[0017] The inbred BALB / c mice were sacrificed, the spleen was removed, and the culture medium was used to make cell suspension. Then 100 μl of cell suspension was implanted into a 96-well culture plate, 90 μl of glycyrrhizin of different concentrations was added, and 10 μl of mitogen ConA was added at the same time, and 90 μl of culture solution containing 10% serum was added to the control group. cultured for 72 hours. At the end of the culture, carefully aspirate the supernatant from each well, then add 10 μl MTT, and continue to incubate in the incubator. After 4 hours, add 100 μl of formazan lysate to each well, put it back into the incubator and incubate overnight, and read OD570 on a microplate reader.
[0018] Analysis of the experimental data shows that glycyrrhizin can significantly inhibit the proliferation of T cells induced by ConA (see figure 1 ), and the ...
Embodiment 3
[0019] Example 3: Glycyrrhizol inhibits one-way mixed lymphocyte reaction (MLR):
[0020] Inbred lines BALB / c and C 57 BL mice were sacrificed, spleens were taken, and cell suspensions were made with culture medium. Among them, the inbred strain C 57 Splenocytes from BL mice were treated with mitomycin C as responder cells, splenocytes from inbred BALB / c mice were used as stimulator cells, and the two cell suspensions were mixed in equal volumes. Then, 100 μl of mixed cell suspension and 100 μl of drug solution were added to the 96-well culture plate, and 100 μl of culture solution containing 10% serum was added to the control group, and the two kinds of cells were cultured separately as a control. Incubate for 72 hours at 37°C, 5% carbon dioxide. At the end of the culture, carefully aspirate the supernatant from each well, then add 10 μl MTT, and continue to incubate in the incubator. After 4 hours, add 100 μl of formazan lysate to each well, put it back into the incubato...
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