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Active dendrobe polysaccharide capable of protecting liver and resisting liver fibrosis and preparation method of antibody affinity chromatography

A technology for anti-liver fibrosis and Dendrobium polysaccharide is applied in the field of plant extract and plant extract preparation, and can solve the problems of low yield, low purity and the like

Active Publication Date: 2012-06-20
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of low yield and low purity of the existing preparation method of Dendrobium Huoshanense polysaccharides, the object of the present invention is to provide an antibody with hepatoprotective and anti-hepatic fibrosis polysaccharides of Dendrobium Huoshanense with high product purity, controllable quality and high purification efficiency Affinity Chromatography Preparative Method

Method used

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  • Active dendrobe polysaccharide capable of protecting liver and resisting liver fibrosis and preparation method of antibody affinity chromatography
  • Active dendrobe polysaccharide capable of protecting liver and resisting liver fibrosis and preparation method of antibody affinity chromatography
  • Active dendrobe polysaccharide capable of protecting liver and resisting liver fibrosis and preparation method of antibody affinity chromatography

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Effect test

Embodiment 1

[0042] Example 1 : Extraction, purification and structural characterization of active dendrobium polysaccharides

[0043] The specific operation steps of the extraction, separation and purification method of the hepatoprotective and anti-hepatic fibrosis active dendrobium polysaccharide are as follows:

[0044] (1) Raw material pretreatment: Take crushed Dendrobium huoshanense, degrease and decolorize with petroleum ether at 50°C for 5 hours, and place it in an environment with a temperature of 25°C to remove the organic solvent by volatilization to obtain the medicinal residue of Dendrobium;

[0045] (2) Hot water extraction: Take Dendrobium medicinal dregs, add distilled water according to the mass-volume ratio of material to liquid 1 g︰30ml, and extract at a temperature of 80°C for 60 minutes to obtain an extract; extract the extract at a speed of 15000 r / min centrifuged for 15 min under the same conditions, and the centrifuged sediment obtained was extracted twice with h...

Embodiment 2

[0055] Example 2 : Antibody affinity chromatography preparation method of active dendrobium polysaccharide

[0056] Step 1 Prepare the immunogen: take the active dendrobium polysaccharide and dissolve it in a carbonate buffer solution with a pH value of 9.2 to prepare an active dendrobium polysaccharide solution with a mass fraction of 10 mg / ml, and then add bovine serum albumin to make the active dendrobium polysaccharide solution The mass fraction of bovine serum albumin in the mixture is 10 mg / ml, stir and mix well, and place it in a thermostat at 37°C for 3 days to obtain a reaction system; add ethanolamine to make the mass fraction of ethanolamine in the reaction system reach 1%. Terminate the reaction to obtain a reaction solution; the reaction solution is separated by a Sephadex G-150 gel column and collected in parts; respectively, the phenol-sulfuric acid method and the Bradford method are used to measure the content of active dendrobium polysaccharides and bovine se...

Embodiment 3

[0064] Example 3 : Protective effect of dendrobium polysaccharides on carbon tetrachloride-induced acute liver injury and liver fibrosis in rats

[0065] After adaptive feeding for one week, SD rats were weighed and randomly divided into 6 groups: normal control group, model control group, low-dose Dendrobium polysaccharide group (100 mg / kg.bw), medium-dose Dendrobium polysaccharide group (200 mg / kg .bw), Dendrobium polysaccharide high-dose group (400 mg / kg.bw), bifendate positive control group (200 mg / kg.bw), 10 rats in each group. Except for the normal control group, all other groups received subcutaneous injection of CCl 4 1. Rat liver fibrosis model induced by peanut oil mixture (1:9), 2 times a week, 4.0 ml each time, for 12 consecutive weeks. From the 5th week, the corresponding drugs (10.0 ml / kg) were fed into the groups respectively, once a day for 9 consecutive weeks. The rats were sacrificed 24 hours after the last administration, the serum was separated, and the...

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Abstract

The invention discloses active dendrobe polysaccharide capable of protecting liver and resisting liver fibrosis, whose chemical structural formula is seen in the figure. The active dendrobe polysaccharide is gala glucomannan taking glucose and mannose as a main chain and galactose as a branch, the molecular weight of the gala glucomannan is about 2.2*104dalton, and the mole ratio of glucose to mannose to galactose in the composition is 31:10:8. The invention further discloses extraction, separation and purification methods of the active dendrobe polysaccharide as well as the preparation method of antibody affinity chromatography of the active dendrobe polysaccharide. The active dendrobe polysaccharide prepared by the method has remarkable physiological activity of protecting liver and resisting liver fibrosis and can be used for preparing liver-protecting health care products and medicines for curing liver fibrosis.

Description

technical field [0001] The invention belongs to the field of plant extracts and preparation methods of plant extracts, and in particular relates to a dendrobium polysaccharide extract with anti-hepatic fibrosis activity and an antibody affinity chromatography preparation method thereof. Background technique [0002] The liver is the most important detoxification organ in the human body. Frequent contact of viral agents, alcohol, drugs and other toxic substances with the liver can lead to liver damage. Liver injury can cause pathological changes such as cell necrosis, apoptosis, inflammation, and fibrosis, and eventually lead to various liver diseases such as liver fibrosis. Liver fibrosis is the common pathological basis for various liver diseases to develop into liver cirrhosis and liver cancer. It is the main cause of death in patients with liver diseases and is extremely harmful to human health and quality of life. At present, the treatment of liver injury mainly relies ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00C07K16/16A61K31/715A61P1/16A23L1/09
Inventor 罗建平潘利华查学强
Owner HEFEI UNIV OF TECH
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