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Flarnmulina velutipes polysaccharide and preparation method thereof

A technology of Flammulina velutipes polysaccharide and Flammulina velutipes crude polysaccharide, which can be applied in the directions of antiviral agents, allergic diseases, drug combinations, etc., can solve problems such as uncertainty of components and biological applications, and achieve easy control of product quality, less allergic reaction, and safety. high effect

Inactive Publication Date: 2017-03-22
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] CN201210219695 discloses a method for extracting Flammulina velutipes polysaccharides by ultrasonic waves, which discloses that Flammulina velutipes polysaccharides have immunomodulatory activity, but only evaluates the in vitro activity through experiments, and does not involve the evaluation of its in vivo activity; CN201410599264 discloses a method through It is a method to improve the purity of Flammulina velutipes polysaccharide by boiling and ethanol precipitation, but the composition and biological application of the polysaccharide extracted by this method cannot be determined, and the obtained polysaccharide is not a pure product

Method used

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  • Flarnmulina velutipes polysaccharide and preparation method thereof
  • Flarnmulina velutipes polysaccharide and preparation method thereof
  • Flarnmulina velutipes polysaccharide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The preparation of Flammulina velutipes polysaccharide comprises the following steps:

[0034] (1) Degreasing: take the fruiting bodies of Flammulina velutipes, dry them in a blast at 60°C to constant weight, crush the dried fruiting bodies of Flammulina velutipes, add 95% ethanol at a material-to-liquid ratio of 1:8 (W / V), and store at room temperature Fully soak for 24 hours, filter to remove lipids;

[0035] (2) Repeat step (1) twice;

[0036] (3) Water extraction: collect the residue, dry it at room temperature, add distilled water according to the ratio of material to liquid 1:20 (W / V), soak it fully, react at 100°C for 1 hour, filter, and collect the filtrate;

[0037] (4) Repeat step (3) twice, and combine the filtrates of the three reactions;

[0038] (5) Concentrate the filtrate, then add absolute ethanol until the mass percent concentration of ethanol reaches 60%, stir well, place in the shade, let stand for 10 hours, centrifuge, and collect the precipitate;...

Embodiment 2

[0046] Preparation of test samples

[0047] Accurately weigh the Flammulina velutipes crude polysaccharide and Flammulina velutipes polysaccharide samples prepared in Example 1 in a sterilized eppendorf tube, configure a solution with a concentration of 5 mg / mL with PBS buffer, and centrifuge at 12000 rpm / min for 30 min after fully dissolving. Transfer to a new sterile eppendorf tube and dilute the sample to 50, 200 and 500 μg / mL for use. The negative control was PBS buffer, and the positive control was 10 μg / mL LPS solution.

[0048] Preparation of mouse RAW264.7 macrophages

[0049] in DMEM medium at 37°C, 5% CO 2 After subculture under conditions, digest with 0.25% trypsin, collect the cells after the suspension is centrifuged at 300g for 3min, and dilute the cells to a certain concentration with colorless RPMI1640 medium for later use.

[0050] Determination of NO release activity of macrophages

[0051] Because NO is extremely unstable, nitrous acid groups (NO2 - ) a...

Embodiment 3

[0060] Fibroblast Chicken Embryo Cell Preparation

[0061] Take 9-10 day-old well-developed chicken embryos and sterilize them with iodine wine and alcohol cotton balls. Under aseptic operation, take out the chicken embryos and put them on a sterilized plate, remove the chicken embryo heads, limbs and internal organs, and rinse the chicken embryos with PBS solution for 3 Second-rate. Transfer the cleaned chicken embryos into a sterile 100mL small beaker, and cut them into 1mm pieces with ophthalmic scissors. 3 The organization is fast, nearly chylous. First wash with PBS solution 3 times, and let it stand for a few minutes, wait for the tissue pieces to sink, absorb and discard the upper liquid, then add PBS solution, transfer to a cell culture bottle, let stand for a few minutes, wait for the tissue pieces to sink, absorb and discard For the upper layer of PBS solution, add 5 mL of 0.25% trypsin solution to the tissue pieces of the cell culture flask, the ratio of trypsin s...

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Abstract

The invention discloses a flarnmulina velutipes polysaccharide and a preparation method thereof. The method includes the preparation steps of hot water extraction, ethanol precipitation, ion-exchange column chromatography and gel column chromatography. The method is simple, easy to implement, safe and reliable, and the flarnmulina velutipes polysaccharide prepared through the method is high in purification concentration, has definite molecular structure characteristics such as molecular weight and monosaccharide composition, contains four monosaccharides including galactose, mannose, fucose and glucose, and can inhibit the Newcastle disease virus and improve the immunocompetence of animals to some extent.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a polysaccharide from Flammulina velutipes and a preparation method thereof. Background technique [0002] Flammulina velutipes is one of the most widely cultivated edible fungi, and its output ranks among the top three in the world. A large number of studies have shown that the main functional component of Flammulina velutipes is Flammulina velutipes polysaccharides, which have various functional activities such as anti-tumor, immune regulation, lowering cholesterol, lowering blood pressure, improving memory, anti-inflammation, anti-virus, antibacterial and anti-oxidation. Flammulina velutipes polysaccharide is a mixed polysaccharide composed of several polysaccharide components mainly composed of dextran. Flammulina velutipes polysaccharides obtained by different extraction and separation methods have different molecular structure characteristics such as molecular weight and monosa...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K31/715A61P31/14A61P37/04
CPCC08B37/006A61K31/715C08B37/0003
Inventor 汪雯翰贾薇张劲松庄海宁刘艳芳杨焱吴迪张赫男
Owner SHANGHAI ACAD OF AGRI SCI
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