Application of Ligustrum glucosides inducing antiviral cytokines
A technology of ligustrum purpurea and cytokines, which is applied in the direction of antiviral agents, medical preparations containing active ingredients, allergic diseases, etc. Report and other issues, to achieve the effect of fast and effective method and wide source of raw materials
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Embodiment 1
[0036] Extraction of Ligustrum glucosides
[0037] Take 1.92kg of dried leaves of Ligustrum adenophorum, add 19.2kg of ethanol, heat to 60°C for reflux extraction three times, dissolve the obtained ethanol extract in water, and separate the water-soluble matter by Diaion column chromatography, and use water- The ethanol mixture was eluted, and the eluate was concentrated to obtain 230.4 g of privetin.
[0038] The obtained privet glucosides are light yellow powder with fragrance, easily soluble in water, and the aqueous solution is light brown.
[0039] The resulting privet glucosides were detected by HPLC. The chromatographic conditions were: the mobile phase was water and methanol containing 0.1% formic acid, water:methanol=90:10~10:90 gradient elution, the time was 60min, and the flow rate was 1ml / min, the injection volume is 10μl, the detection wavelength is 227nm, the chromatogram is as follows figure 1 As shown, the main peak appeared at 7min, 20min, 31min, 38min and ...
Embodiment 2
[0042] Experiment of Inducing Lymphocytes and Macrophages to Secrete Cytokines in Mouse Lymphocytes and Macrophages In Vitro
[0043] 1. Lymphocyte Preparation and Purification
[0044] (1) Execute the mouse by neck dislocation, then put the mouse into 75% alcohol and soak it for 5 minutes, so that its skin contacts alcohol as much as possible;
[0045] (2) Put the sterilized mouse into the ultra-clean workbench, cut open the left abdomen, take out the spleen, put it into the pre-cooled serum-free RPMI-1640 culture medium (3-5ml), remove the fat and connective tissue, and then Wash with pre-cooled serum-free medium;
[0046] (3) Cut the cleaned mouse spleen into pieces, grind it with a glass syringe plug, pass it through a 200-mesh nylon mesh, and rinse the nylon mesh with pre-cooled serum-free medium;
[0047] (4) Draw the upper lymphocyte suspension into the test tube, add 4-5ml mouse lymphocyte separation medium, and centrifuge at 800rcf (relative centrifugal force) for 3...
Embodiment 3
[0061] Cytokine secretion in immunocompromised mice induced by privet glucosides (LPGs) in vivo
[0062] 1. Randomly group Kunming mice, 12 in each group, respectively: blank control group: normal saline 10ml / kg; model group: normal saline 10ml / kg; levamisole hydrochloride group: 50mg / kg; High-dose group: 800mg / kg; medium-dose group: 400mg / kg; low-dose group: 200mg / kg. The mice in each group were given continuous intragastric administration for 10 days, starting from the first day of administration, the blank control group was intraperitoneally injected with sterilized normal saline 10ml / kg every other day, and the other groups were intraperitoneally injected with cyclophosphamide 80mg / kg every other day, a total of 5 times.
[0063] 2. Determination of IFN-β and IFN-γ levels in peripheral blood by ELISA experiment, the results are as follows Figure 9 shown.
[0064] Depend on Figure 5 It can be seen that cyclophosphamide can cause down-regulation of IFN-β and IFN-γ in se...
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