Anti-hepatitis B virus streblus extract and extracting technique thereof
An extract and anti-hepatitis B technology, applied in the direction of antiviral agents, drug combinations, plant raw materials, etc., to achieve the effect of mature separation technology, rich resources, and powerful effects
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Embodiment 1
[0017] (1) Extraction: Get 2 kilograms of naturally air-dried magpie kidney branches and leaves, heartwood, root, and bark, and use 75% ethanol to reflux after crushing, (3 × 30 liters), combine the extracts, recycle the solvent after filtration, and obtain red 1000 ml of the concentrated solution was placed in a 5000 ml separating funnel and extracted with petroleum ether, diethyl ether, ethyl acetate and n-butanol respectively. After recovering the solvent, 0.8 g of petroleum ether extract, 3.2 g of diethyl ether extract, acetic acid 33 grams of ethyl ester extract, 38 grams of n-butanol extract, and 87 grams of water-soluble matter.
[0018] (2) Separation:
[0019] Take 3.2 g of the ether extract and carry out silica gel column chromatography (300-400 mesh, 200 g), and use MeOH:CHCL 3 (0:100→5:95) gradient elution, five components were obtained, the weights were 0.3g, 0.6g, 0.5g, 0.2g and 1.2g, and each component was subjected to preparative thin-layer chromatography , e...
Embodiment 2
[0023] The effect of n-butanol extract of magpie kidney tree on the expression of HepG2.2.15, cell lines HBsAg and HbeAg:
[0024] Inoculate HepG2.2.15 cells 2×105 / ml into 96-well cell culture plate, 190ul per well, add the drug concentration of 800ug / ml, 400ug / ml, 200ug / ml, 100ug / ml into the cell culture plate, each concentration Set 3 wells, set 10% DMSO group, 3 wells are used as negative control, take the medium supernatant on the 3rd day and 6th day respectively, and store it at -20°C, and use the ELISA method to measure HBsAg and HbeAg at a wavelength of 450nm The absorbance at the place, the inhibition rate of the drug on the cells was calculated according to the following formula.
[0025]
[0026] See Table 1 and Table 2 for the results of the inhibition rate of the drug on the secretion of HbsAg and HbeAg:
[0027] Table 1 Suppressive effect of Que Kidney tree extract on HepG2.2.15 cell line HbsAg expression (X±D, n=3)
[0028]
[0029] The inhibitory ...
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