Application of orientin in the preparation of drugs for inhibiting angiogenesis
An angiogenesis and orientin technology, applied in the field of biomedicine, can solve the problems of undiscovered orientin patents and literature, and achieve the effect of convenient extraction and purification, low cytotoxicity, and inhibition of angiogenesis
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Embodiment 1
[0021] Preparation of orientin
[0022] 1. Dry 3.0 kg of Euphorbia euphorbia whole herb, pulverize it, extract it by cold soaking with 95% ethanol for 3 times (7 days each time), recover the solvent to obtain 600 g of ethanol extract, add deionized water to dissolve, and then use petroleum ether , ethyl acetate, and n-butanol extraction to obtain 70 g of petroleum ether extract, 50 g of ethyl acetate extract, and 62 g of n-butanol extract.
[0023] 2. Add an equal amount of silica gel to 50 g of ethyl acetate extract and mix the sample, then carry out silica gel column chromatography, and use petroleum ether: ethyl acetate gradient elution (50:1~0:100); thin layer chromatography (TLC) tracking, combined The ratio of petroleum ether: ethyl acetate is 10:1~5:1 fraction, the pigment is removed by MCI, and then separated by silica gel column chromatography to obtain 20 mg of yellow powder.
[0024] Spectral detection data of the yellow powder: ESI-MS (m / z) 447 [M-H] - . 1 H-NMR...
Embodiment 2
[0027] The inhibitory effect of orientin on angiogenesis was confirmed by the experiment of the inhibitory effect of orientin on human umbilical vein endothelial cells (human vascular endothelial cells, HUVEC).
[0028] Tetrazolium salt (MTT) colorimetric method was used to observe the inhibitory effect of orientin on the proliferation of HUVEC in the positive control group arbutin (UA) group and low, medium and high dose groups, indicating that orientin had a good effect on inhibiting the proliferation of HUVEC. And there is a concentration-dependent relationship.
[0029] (1) HUVEC preparation, grouping and processing
[0030] Digest monolayer cultured cells with 0.25% trypsin, prepare single cell suspension with DMEM medium containing 10% newborn bovine serum, and cells in 5.0×10 4 Cells / mL were inoculated in a 96-well culture plate, 200 μL per well; about 90% of the cultured cells were confluent and replaced with serum-free DMEM culture medium for 24 hours; ), positive c...
Embodiment 3
[0040] Effect of orientin on the migratory ability of HUVEC.
[0041] The cell scratch healing method is one of the methods for measuring the cell movement characteristics; the experimental results show that the positive control group is different from the low, middle and high dose groups. Experiments have confirmed that orientin has the effect of inhibiting the proliferation of HUVEC. In the high-dose group of orientin, there are almost no HUVECs in the scratches, and the inhibitory effect is very obvious.
[0042] In this embodiment, the blank group (DMEM culture fluid), the negative control group (0.1% DMSO), the positive control group (UA50 μ g / mL) and the low, middle and high dose groups of orientin are set with the same embodiment 2, but The orientin medicament used is the orientin 50wt% medicament prepared from the orientin prepared in Example 1.
[0043] (1) HUVEC preparation
[0044] Digest monolayer cultured cells with 0.25% trypsin, prepare single cell suspension ...
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