Pregnane glycoside compound and application thereof
A kind of technology of pregnane glucoside and compound, applied in the field of pregnane glucoside compound
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Embodiment 1
[0020] Example 1: Preparation of Epigycoside B
[0021] Sorting the branches and leaves of Epigynum cochinchinensis (Epigynum cochinchinensis), and drying 5 kg of the obtained leaves naturally in a dry and ventilated place, and crushing them into pieces using a powder machine; 12 hours, continuous leaching three times; collect and combine the extracts, use a rotary evaporator to concentrate under reduced pressure to remove the liquid, and obtain 500g of extract; add 2L of water to dissolve the extract, put it in a separatory funnel, and add an equal volume of ethyl acetate Carry out repeated extraction; Repeated extraction until the ethyl acetate layer is almost colorless, collect and combine the ethyl acetate layer and evaporate to dryness to obtain 120 g of the crude extract extract; use a medium-pressure chromatographic separation system to pass the crude extract Wash 5 column volumes with 90% methanol water to obtain a sample free of plant pigments and other impurities, an...
Embodiment 2
[0031] Embodiment 2: Immunosuppression detection test
[0032] (1) Preparation of spleen lymphocyte suspension
[0033] 18-22g healthy BABL / c mice were killed by removing the eyeballs and bloodletting, soaking them in alcohol with a volume concentration of 75% for 3-5 minutes for disinfection, taking them out and placing them in a sterile tray with the left side of the abdomen facing upwards. In the cabinet, use sterilized tweezers to pick up the fur in the middle of the abdomen, make an incision from the lower end, use another set of instruments to cut open the layers of the abdominal wall, use the third set of instruments to remove the spleen, remove fat and connective tissue, and put it in PBS (Phosphate buffered saline) to wash away floating blood. Then move the spleen tissue to a plate filled with RPMI1640 incomplete culture medium, cut it into small pieces with scissors, use a sterile syringe core to gently grind the spleen in a 200-mesh stainless steel screen, wash it ...
Embodiment 4
[0061] Embodiment 4: Anti-inflammatory activity test in vitro
[0062] (1) Preparation of test-related reagents
[0063] Preparation of sample dilution working solution: The concentrated sample dilution solution was taken out from the -4°C refrigerator, thawed slowly at room temperature, and diluted according to the amount required for the test with double distilled water at a ratio of 1:5.
[0064] Preparation of reagent dilution working solution: take the concentrated reagent solution out of the -4°C refrigerator, thaw slowly at room temperature, take part according to the amount required for the test, and dilute it with double distilled water at a ratio of 1:5.
[0065] Preparation of cell lysis working solution: The concentrated cell lysate was taken out from the refrigerator at -4°C, thawed slowly at room temperature, and diluted with double distilled water at a ratio of 1:2 according to the amount required for the test.
[0066] Preparation of washing working solution: ...
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