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Acidic aloe polysaccharide as well as preparation and purification method and application thereof

An aloe polysaccharide and acidic technology, which is applied in the direction of pharmaceutical formulas, organic active ingredients, and medical preparations containing active ingredients, etc., can solve the problems of increased incidence of drug-resistant strains, treatment failure, etc., achieve high yield and reduce adhesion rate , equipment requires simple effect

Inactive Publication Date: 2010-06-09
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current treatment for Helicobacter pylori mainly uses triple therapy, but with the widespread use of antibiotics, the incidence of drug-resistant strains has gradually increased, leading to treatment failure

Method used

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  • Acidic aloe polysaccharide as well as preparation and purification method and application thereof
  • Acidic aloe polysaccharide as well as preparation and purification method and application thereof
  • Acidic aloe polysaccharide as well as preparation and purification method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Preparation and purification of embodiment 1 acidic aloe polysaccharide

[0026] (1) Fresh aloe leaves are chopped, extracted with hot water, and precipitated with ethanol to obtain a crude extract. Weigh an appropriate amount of crude extract, soak it twice with 50-70% ethanol for 24-48 hours each time, after the residue evaporates to dry the solvent, dissolve it in an appropriate amount of distilled water with stirring, and centrifuge to remove the insoluble matter. Trichloroacetic acid was added to the supernatant to remove protein, and after standing overnight, the supernatant was collected by centrifugation, and the above operations were repeated. After the supernatant was concentrated under reduced pressure, 95% ethanol was added to stand overnight and then centrifuged. After the precipitation was dried, a white or light yellow blocky solid was obtained, which was the aloe crude polysaccharide.

[0027] (2) Aloe crude polysaccharides were purified by ion exchange...

Embodiment 2

[0028] Example 2 Inhibitory effect of acidic aloe polysaccharides on Helicobacter pylori adhesion to MKN 45 cells

[0029] (1) Helicobacter pylori was streaked and inoculated on Columbia agar medium (containing 7% fresh sheep blood), and cultured at 37° C. for 3 days in a microaerophilic environment. Afterwards, the colonies were scraped and placed in sterilized physiological saline, and the bacterium-adjusting solution was at an appropriate concentration. Add FITC-DMSO solution to the prepared bacterial solution, and incubate at room temperature for 1-3 hours. Then centrifuge the marked bacterial suspension, pour out the supernatant, add D-Hanks solution containing Tween-20 and pipette several times, continue centrifuging, repeat the above operation twice, and wash off the unmarked FITC.

[0030] (2) Discard the original culture medium of MKN45 cells cultured in the 96-well plate, and replace with 2% calf serum and 1640 culture medium containing acidic aloe polysaccharides o...

Embodiment 3

[0032] Example 3 Effect of acidic aloe polysaccharides on Escherichia coli adhesion to LOVO cells

[0033] LOVO cells were cultured in 1640 medium containing 10% calf serum for 48 hours, and then cultured in 96-well plates. Escherichia coli was cultured in LB medium for 24 hours, and the colonies were scraped and labeled with FITC according to the above method. Discard the original culture medium of LOVO cells cultured in the 96-well plate for 24 hours, and add acidic aloe polysaccharides with the contents of 0, 0.01, 0.1, 0.5, 1.0mg / ml and 2% calf serum to each group of 12 wells 1640 medium, incubate for 1h. After adding FITC-labeled Escherichia coli and incubating for 1 h, the unadhered bacteria were washed away, and the fluorescence value was measured with a microplate reader. see results image 3 .

[0034] image 3 The results showed that, compared with the control group, the adhesion rate of Escherichia coli to LOVO cells decreased to 78% (p<0.001) and 68% (p<0.001) r...

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Abstract

The invention belongs to the technical field of biological pharmacy, in particular to a preparation and purification method of acidic aloe polysaccharide and application thereof in a helicobacter pylori and escherichia coli resistant medicament. A test proves that the method for obtaining acidic aloe polysaccharide by adopting Chinese aloe as a raw material, extracting by hot water, obtaining a rough extract by an ethanol precipitation method, dissolving by hot water, removing protein by trichloroacetic acid, obtaining rough polysaccharide after ethanol precipitation and purifying by a DEAE-52 ion-exchange column chromatography has the characteristics of high product purity, higher yield, simple equipment requirement and more favorable repeatability (shown as a summary attached figure). The acidic aloe polysaccharide can effectively reduce the sticking rate of helicobacter pylori on an MKN 45 cell. In addition, the acidic aloe polysaccharide also has the obvious inhibiting function on sticking an LOVO cell by escherichia coli. The acidic aloe polysaccharide is a broad-spectrum sticking resistant agent and can be used for preparing a helicobacter pylori sticking resistant medicament and applied to treating relevant diseases caused by the infection of the helicobacter pylori.

Description

1. Technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to the preparation and purification of acidic aloe polysaccharide and its application in the preparation of anti-helicobacter pylori adhesion drugs and the treatment of related diseases caused by helicobacter pylori. 2. Background technology [0002] Aloe vera has been widely used as a medicinal and edible plant since ancient times. Modern pharmacological studies have shown that aloe vera has the functions of sterilization, anti-inflammation, diarrhea, anti-oxidation, anti-radiation, anti-ulcer and anti-tumor. According to research reports at home and abroad, there are two types of active ingredients in aloe vera, one is anthraquinones such as aloin that exist at the bottom of aloe leaves and near the epidermis, and the other is anthraquinones from fresh leaves of aloe parenchyma About 98.5% of the aloe vera gel isolated from the natural water is natural w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00A61K31/715A61P31/04
CPCY02A50/30
Inventor 徐琛阮小明李洪森郭彬歆
Owner NANJING UNIV
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