Preparation method as well as structural partial characterization and application of gelidium pacificum okam polysaccharide with immunomodulatory effect

A polysaccharide and immunomodulatory technology of lilyflower, which is applied in the field of natural macromolecules to achieve the effects of promoting immune recovery, inhibiting inflammation and improving human immunity.

Active Publication Date: 2018-06-15
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preparation, purification and physiological activity of the Dashi cauliflower polysaccharide of the present invention have not been reported at home and abroad.

Method used

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  • Preparation method as well as structural partial characterization and application of gelidium pacificum okam polysaccharide with immunomodulatory effect
  • Preparation method as well as structural partial characterization and application of gelidium pacificum okam polysaccharide with immunomodulatory effect
  • Preparation method as well as structural partial characterization and application of gelidium pacificum okam polysaccharide with immunomodulatory effect

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Experimental program
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Effect test

Embodiment 1

[0043] 1. Extraction process

[0044] The dried dashi cauliflower is removed by hand, soaked to remove impurities, dried, crushed, and passed through a 20-mesh sieve. The dashi cauliflower powder is first soaked for 20 minutes, and the material-to-liquid ratio is 1:30. Ultrasound-assisted extraction at 80°C for 2.5 hours to obtain a water extract, 60 °C and concentrated under reduced pressure to obtain a concentrated solution.

[0045] Add twice the volume of absolute ethanol to the concentrated solution, let it stand at room temperature for 12 hours to precipitate polysaccharides, centrifuge at 8000 rpm for 10 minutes, and collect the precipitates at 4°C to obtain crude polysaccharides.

[0046] The crude polysaccharide was dissolved in distilled water, protein was removed by Sevage method, trichloroacetic acid with a final concentration of 3% was added to the concentrated solution, left standing for 12 hours, centrifuged at 8000 rpm for 10 minutes, and the supernatant was ta...

Embodiment 2

[0062] Materials and reagents: Dashi cauliflower polysaccharide A-11 (self-made), DMEM medium (Sigma), LPS (Sigma), fetal bovine serum (Sigma), Griess reagent (Sigma).

[0063] Instrument constant temperature incubator (Shanghai Boxun Industrial Co.), microplate reader (CORONA, Japan), microscope (Shanghai Optical Instrument Factory), 96-well plate (BD, USA).

[0064] 1. The effect of polysaccharides on cell viability

[0065] Cell viability was determined by MTT method. THP-1 cells were divided into six groups with 3 parallels in each group: a blank group was added with 200 μL RPMI-1640 culture solution; a positive control group was added with RPMI-1640 culture solution containing 2 μg / mL LPS; four test groups were added with 2 μg / mL LPS mL LPS and RPMI-1640 culture solution with different concentrations of polysaccharides (1, 5, 10 and 25 μg / mL); cultivated under these conditions for 24 hours. Centrifuge at 1000r / min for 5 minutes to collect cells, add 20 μL of MTT solutio...

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Abstract

The invention relates to gelidium pacificum okam polysaccharide as well as a preparation method and application thereof, and belongs to the field of natural polymers. The invention provides a method for extracting and separating polysaccharide from gelidium pacificum okam, as well as a chemical structure and application of the polysaccharide. The average relative molecular weight is determined byutilizing GPC and the monosaccharide composition is analyzed by using GC-MS; the average relative molecular weight of the gelidium pacificum okam polysaccharide A-11 disclosed by the invention is found to be 28807Da and the gelidium pacificum okam polysaccharide A-11 is mainly prepared from xylose and galactose; in addition, trace amounts of rhamnose, arabinose, fucose, allose, fructose, glucose and mannose are found. Meanwhile, the gelidium pacificum okam polysaccharide A-11 disclosed by the invention has the advantages that the cytotoxicity of LPS-induced THP-1 is significantly inhibited, sothat the cells are prevented from being damaged; besides, the expression of signal factors MyD88 and TRAF-6 in an immune signaling pathway can be regulated so as to enhance human immunity.

Description

technical field [0001] The invention relates to the technical field of natural polymers, in particular to a preparation method, structural part characterization and application of Dashiliu polysaccharide. Background technique [0002] Agaricus, also known as sea jelly, onyx, Qiongzhi, grass coral, chicken feathers, etc., is mainly distributed in Taiwan Island, Hainan Island and the coast of Guangdong in my country. It is currently the main raw material for making agar. According to the records of traditional Chinese medicine, agaricus can be used to treat bronchitis, goiter, enteritis, hemorrhoids, etc. It is a kind of algae with both medicine and food. At present, the main utilization methods of agaricus are direct consumption and production of agar gum, which are widely used in food and chemical industries. There have been some studies on agaricus polysaccharides, and studies have shown that agaricus polysaccharides have good biological activities, such as antioxidant act...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00A61K31/715A23L33/125A61P37/02A61P35/00
CPCA23L33/125A61K31/715C08B37/0003C08B37/006
Inventor 刘克海崔明晓邹颖邬军文张敏王玉
Owner SHANGHAI OCEAN UNIV
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