Euphorbiaceae plant active substance, and preparation method and application thereof
An active substance, technology of Euphorbiaceae, applied in the field of preparation of Euphorbiaceae plant active substances and their active ingredients, can solve the problems of animal husbandry loss, food safety concerns, unsatisfactory effect, etc., and achieve the effect of inhibiting proliferation
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Embodiment 1
[0043] Example Preparation of Euphorbiaceae Plant Active Substances
[0044] 1. Active substances extracted by ethanol (Ethanol)
[0045] Euphorbia neriifolia was collected from Yilan, Taiwan. First, wash the collected diamonds with water and separate the leaves and stems. After standing to dry, weigh them and cut them into slices. Then put the stems and leaves in a brown glass bottle that can protect from light, add 95% (volume percent concentration, 50-99% is acceptable) ethanol to completely immerse the stems and leaves of King Kong in ethanol, and stand for extraction at room temperature until the ethanol changes from transparent and colorless to dark green or dark brown (about 3 -10 days), the ethanol extract was collected, and then concentrated under reduced pressure to remove the ethanol, to obtain the active substances of the stem and leaves of the rhizome extracted with ethanol.
[0046] 2. Active substances extracted by n-hexane (Hexane)
[0047] Dissolve 10 g of th...
Embodiment 2
[0052] Example 2 Preliminary activity analysis of Euphorbiaceae plant active substances extracted with ethanol against PRRSV virus infection
[0053] The active substances in the stem and leaves of the ethanol extracted from Example 1 were first serially diluted twice (500 μg / mL, 250 μg / mL, 125 μg / mL, 62.5 μg / mL, 31.25 μg / mL, 15.625 μg / mL) μg / mL, 7.8125 μg / mL, 3.90625 μg / mL, 1.953125 μg / mL, 0.9765625 μg / mL), followed by treatment of Marc-145 cells (epithelioid cells, derived from monkey kidney cells, derived from mother cells (MA 104 cells) The cloned and obtained PRRSV virus was re-infected to evaluate whether the ethanol-extracted adamantine active substance had the ability to inhibit PRRSV virus infection.
[0054] First, the Marc-145 cells were suspended in MEM medium containing 5% FBS, and then seeded in 96-well cell culture dishes (1.5×10 4 cells / well) at 37°C, 5% CO 2 After culturing cells in a constant temperature incubator for 16-24 hours, remove the supernatant med...
Embodiment 3
[0058] Example 3 Activity analysis of the active substances from the stems of Euphorbiaceae plants extracted with ethanol under different action times against PRRSV virus infection
[0059] The ethanol-extracted adamantium stem active substance (2 μg / mL) obtained in Example 1 was treated with Marc-145 cells for different times and then infected with PRRSV virus to evaluate the relationship between the ethanol-extracted adamantha stem active substance and Marc-145 How long the cells are co-cultured will have the best ability to inhibit PRRSV virus.
[0060] First, the Marc-145 cells were suspended in MEM medium containing 5% FBS and seeded in a 96-well cell culture dish (1.5×10 4 cells / well) at 37°C, 5% CO 2 After culturing the cells in a constant temperature incubator for 16-24 hours, remove the supernatant medium and wash the cells twice with MEM medium containing 1% FBS, then add 2 μg / mL ethanol-extracted active substances from the stems of adamantium (experimental group) ...
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