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A kind of yew polysaccharide and its preparation method and application

The technology of yew polysaccharide and yew is applied in the field of effective utilization of yew resources, which can solve the problems of waste of resources, environment, pollution, destruction of yew resources and the like, and achieve the effect of improving the extraction rate of polysaccharides

Active Publication Date: 2017-09-26
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although paclitaxel has a good anti-cancer effect, because the content of paclitaxel in yew is very low, and it is mostly distributed in the bark, some manufacturers cut down the yew in order to increase the output of paclitaxel, which seriously damaged the resources of yew.
In addition, the yew after paclitaxel is extracted is basically discarded, causing serious waste of resources and environmental pollution

Method used

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  • A kind of yew polysaccharide and its preparation method and application
  • A kind of yew polysaccharide and its preparation method and application
  • A kind of yew polysaccharide and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Extraction and identification of yew polysaccharides

[0051] 1. Extraction of Taxus polysaccharides

[0052] A kind of extraction method of yew polysaccharide of the present embodiment, comprises the following steps:

[0053] (1) Degreasing:

[0054] Take 10.0g of dried yew branches and leaves, put them in a 250mL Soxhlet extractor and extract with petroleum ether with a boiling point of 60-90°C under reflux, and the temperature of the water bath is 80°C; The oil stain is the end point of degreasing, and the degreased yew branches and leaves are left at room temperature to be ventilated and dried, and accumulated together for future use.

[0055] (2) Extraction:

[0056] ①Use water to extract the yew branch and leaf debris first by microwave (power 900W, extraction time 45s, the number of extractions is three times, the interval between two adjacent extractions is 5min or the next extraction is performed after the system cools down), and then placed in 95 ...

Embodiment 2

[0113] Example 2 In vitro anti-tumor activity experiment of CPTC-2

[0114] 1. MTS method detection

[0115] SGC-7901 cells in the exponential growth phase were digested with 0.25% trypsin to make a cell suspension, inoculated on a 96-well culture plate, 100 μL per well, placed in 5% CO 2 , and cultivated in an incubator at 37°C for 24 hours; for the CPTC-2 experimental group, add yew polysaccharides prepared with serum-free RPMI-1640 basal culture medium to the 96-well culture plate, 100 μL per well, to make the final concentration of polysaccharides 0, 12.5, 25, 50, 100, 150, 200, 300 μg / ml, respectively, and set blank control, negative control and astragalus polysaccharide (50 μg / ml) positive control at the same time, and each group had 3 replicates; Add 20 μL of MTS (5 mg / mL) prepared in PBS to the wells, continue to incubate for 2 h, measure the OD value at a wavelength of 490 nm with a microplate reader, and calculate the inhibition rate (RI) according to the formula: ...

Embodiment 3

[0123] Example 3 Anti-tumor synergistic effect of CPTC-2 combined with 5-FU

[0124] 1. MTS method detection

[0125] The experiment set up control group, 5-FU group (50 μg / mL), CPTC-2 high, medium and low dose groups (300, 150, 50 μg / mL) and CPTC-2 high, medium and low (300, 150, 50 μg / mL) mL)+5-FU (50 μg / mL) group, with 3 repetitions in each group.

[0126] SGC-7901 cells in the exponential growth phase were digested with 0.25% trypsin to make a cell suspension, inoculated on a 96-well culture plate, 100 μL per well, placed in 5% CO 2 , Cultivate in an incubator at 37°C for 24 hours, add each group of working solutions, continue to cultivate for 48 hours, add 20 μL of MTS (5 mg / mL) prepared in PBS to each well, continue to incubate for 2 hours, and directly measure OD with a microplate reader at a wavelength of 490 nm value, the inhibition rate (RI) was calculated according to the formula:

[0127] Inhibition rate (RI)=(OD value of the control group-OD value of the experi...

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Abstract

The invention discloses a yew polysaccharide, a preparation method and application thereof. The preparation method includes separation and purification. The separation and purification include: using NaCl solution as an eluent, using a DEAE sepharose fast flow resin column to separate the crude polysaccharide, and obtaining a target fraction eluted with a concentration of 0.3M NaCl solution; using deionized water As the eluent, Sephdex G-10 gel column was used to separate the target fraction, and the yew polysaccharide was obtained when the retention time was 30min. The weight-average molecular weight of the yew polysaccharide is 73527Da, and it is composed of glucose, xylose, mannose, arabinose, rhamnose and galactose in a molar ratio of 1:0.38:0.31:3.11:1.34:1.92; the main chain is (1 →3)‑Arabinose and (1→4)‑Galactose as the backbone have significant apoptosis-inducing effects on gastric cancer cell line SGC‑7901.

Description

technical field [0001] The invention relates to a technology for effectively utilizing yew resources, in particular to a yew polysaccharide and its preparation method and application. Background technique [0002] As early as 1856, Lucas extracted the powdered alkaline part of taxine from the leaves of European yew (Taxus baccata), but the progress was very slow. It was not until the rapid development of spectroscopic technology in the 1960s that there was more in-depth research. In the 1980s, scientists in the United States and Europe successively revealed the anticancer efficacy of paclitaxel, and the whole world set off a climax of research on yew. Although paclitaxel has a good anti-cancer effect, because the content of paclitaxel in yew is very low and it is mostly distributed in the bark, some manufacturers cut down the yew in order to increase the output of paclitaxel, which seriously damaged the resources of yew. . In addition, the yew from which paclitaxel has bee...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00A61P35/00
Inventor 吴绵斌张菲菲林建平杨立荣
Owner ZHEJIANG UNIV
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