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Prunella vulgaris polysaccharide and its preparation method and use

A technology of Prunella vulgaris and polysaccharide, which is used in drug combination, allergic diseases, etc.

Inactive Publication Date: 2016-03-30
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there has been no report of finding polysaccharides with complement inhibitory effect from Prunella vulgaris

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1. Preparation of homogeneous polysaccharides PW-PS1 and PW-PS2

[0025] The medicinal material of Prunella vulgaris is crushed, extracted by percolation with 95% ethanol at room temperature until it is colorless, and the residue evaporates to remove the alcohol smell. After drying, it is boiled in boiling water for 3 times, adding 4 times the volume of water each time, and the extraction time is 2 hours each time. The water extracts obtained by each extraction and filtration were combined, concentrated to a small volume, and precipitated with 4 times the volume of absolute ethanol. After the precipitate was dissolved in a small amount of water, it was deproteinized with 15% trichloroacetic acid at 4°C. The solution was centrifuged to remove the precipitate, the supernatant was adjusted to neutral with 1mol / L NaOH, and dialyzed against flowing water for 3 days. The solution in the bag was concentrated to an appropriate volume, and freeze-dried to obtain the c...

Embodiment 2

[0028] Example 2. Anti-complement classical pathway test in vitro

[0029] Take 0.1ml of complement (guinea pig serum), add barbiturate buffer solution (BBS) to prepare a 1:5 solution, and double-dilute with BBS to 1:10, 1:20, 1:40, 1:80, 1: 160, 1:320 and 1:640 solutions. Take 1:1000 hemolysin, 0.1ml of each concentration of complement and 2% sheep red blood cells (SRBC) and dissolve them in 0.3ml of BBS, mix well, put them into a low-temperature high-speed centrifuge after 30 minutes in a 37°C water bath, and put them into a low-temperature high-speed centrifuge at 5000rpm and 4°C. Centrifuge for 10min. Take 0.2ml of the supernatant from each tube and place it in a 96-well plate, and measure its absorbance at 405nm. At the same time, a complete hemolysis group (0.1ml 2% SRBC dissolved in 0.5ml triple distilled water) was set up in the experiment. The absorbance of three-distilled water lysed blood vessels was used as the standard of total hemolysis, and the hemolysis rate...

Embodiment 3

[0030] Example 3. Anti-complement alternative pathway test in vitro

[0031] Take 0.2ml of complement (human serum), add AP to dilute (barbital buffer, pH=7.4, containing 5mMMg 2+ , 8mMEGTA) liquid was prepared as a 1:5 solution, and double-diluted into 1:10, 1:20, 1:40, 1:80, 1:160, 1:320 and 1:640 solutions. Take 0.15ml of complement of each concentration, 0.15ml of AP diluent and 0.20ml of 0.5% rabbit erythrocytes (RE), mix well, place in a low-temperature high-speed centrifuge after 30 minutes in a 37°C water bath, and centrifuge at 5000rpm and 4°C for 10 minutes. Take 0.2ml of the supernatant from each tube and place it in a 96-well plate, and measure the absorbance at 405nm. At the same time, a complete hemolysis group (0.20ml0.5%RE dissolved in 0.3ml triple distilled water) was set up in the experiment. The absorbance of three-distilled water lysed blood vessels was used as the standard of total hemolysis, and the hemolysis rate was calculated. Taking the dilution of...

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PUM

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Abstract

The invention belongs to the field of Chinese medicine, relates to Prunella vulgaris polysaccharide, and a preparation method and purpose thereof in preparing anticomplement medicines. The Prunella vulgaris polysaccharide including uniform polysaccharide PW-PS1 and PW-PS2 is prepared from a dry fruit aqueous extract of a labiatae plant Prunella vulgaris Linn. Through in vitro tests, the uniform polysaccharide is proved to have strong anticomplement activity and inhibition effects on classical pathway and alternative pathway of a complement system; besides, the polysaccharide does not have the anticoagulant effect influencing in vivo activity and can be used for the preparation of complement inhibition medicaments. The PW-PS1 effects on C1q, C3 and C9 components of the complement system, and the PW-PS2 effects on the C1q, C2, C3, C5 and C9 components of the complement system.

Description

technical field [0001] The invention belongs to the field of traditional Chinese medicine pharmacy, and relates to a Prunella vulgaris polysaccharide and its preparation method and use, in particular to a Prunella vulgaris homogeneous polysaccharide, its preparation method and its use in preparing anti-complement drugs. Background technique [0002] It is known that the complement system is one of the important immune defense systems of the human body. The normal activation of the complement system plays an important role in eliminating foreign microorganisms, removing damaged or dead cells and tissues in the body, and maintaining the balance of the body; however, the complement Excessive activation of the system will cause damage and inflammatory response to the body's own normal tissues. Studies have shown that diseases related to excessive activation of complement include systemic lupus erythematosus, rheumatoid arthritis, acute respiratory distress syndrome and other majo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00A61K31/715A61P37/02
Inventor 程志红陈道峰杜冬生
Owner FUDAN UNIV
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