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Setting-up method for cell membrance solid phase chromatography model for sieving Chinese medicine platelet aggregation resisting active component

An anti-platelet aggregation and active ingredient technology, applied in the direction of material separation, analysis of materials, material inspection products, etc., can solve the problems of easy aggregation, difficult to maintain platelet activity, high cost, etc., achieve good application prospects, low cost, and save money and the effect of time

Inactive Publication Date: 2008-10-29
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, using PMIC to establish an experimental model for screening active ingredients of drugs with antithrombotic effect requires a large number of platelets for the experiment, and the activity of platelets is particularly critical to the experimental results. At present, platelets from animals such as pigs and dogs are mostly used for experiments, and the cost is high. It is difficult to maintain the activity of a large number of platelets collected at one time
The platelet concentration in chicken blood is higher than that of mammals, its function is similar to that of mammals and its activity is stronger, and it is easy to aggregate in vitro, so it is an ideal source of highly active platelets. See the report

Method used

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  • Setting-up method for cell membrance solid phase chromatography model for sieving Chinese medicine platelet aggregation resisting active component
  • Setting-up method for cell membrance solid phase chromatography model for sieving Chinese medicine platelet aggregation resisting active component
  • Setting-up method for cell membrance solid phase chromatography model for sieving Chinese medicine platelet aggregation resisting active component

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Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Method for Establishing Cell Membrane Solid Phase Chromatography Model for Screening Active Components of Traditional Chinese Medicine

[0037] Such as Figure 10 Shown, the chicken cell membrane solid phase chromatography model experiment route of the present invention is as follows:

[0038] (1) Research and analysis of ginger oleoresin test solution and chicken platelet cell membrane immobilization

[0039] (1) Establishment of the fingerprint of the Chinese medicine ginger oleoresin to be screened for the test solution: Accurately weigh 5 mg of ginger oleoresin, dissolve it with 2% Tween-80, and dissolve it with a vortex mixer to obtain the ginger oleoresin test solution. The aqueous phase was filtered through a membrane, and the filtrate was analyzed by HPLC. The analysis conditions are as follows: Chromatographic column: Agilent Zorbax Eclipse XDB-C18 150×4.6 mm, 5 μm. Chromatographic conditions: Gradient elution, mobile phase: A is water, B is aceto...

Embodiment 2

[0054] Embodiment two: the separation and extraction of gingerol

[0055] The present invention extracts gingerol by chemical method and is used as pharmacological experiment to verify the anti-platelet aggregation activity of gingerols, the effect component obtained in the test solution of ginger oleoresin screened by chicken platelet cell membrane solid-phase chromatography established by the present invention. .

[0056]Commercially available ginger was identified by Professor Huang Xuesong from the Department of Food Science and Engineering, College of Science and Technology, Jinan University. Wash the fresh ginger from the soil, cut it into slices, and dry it in a ventilated and dry place, then grind it into 100-mesh ginger powder, and extract the dried ginger powder with acetone; each batch of dried ginger powder is extracted three times; the three extractions are collected together. The solvent is recovered at a temperature lower than 35°C to obtain a crude extract of ...

Embodiment 3

[0057] Embodiment 3: Anti-platelet aggregation experiment of gingerol in vivo

[0058] The mice were randomly divided into three groups, namely solvent control group, aspirin group and gingerol group. Gingerol group was administered with 75 mg / kg dose, aspirin group was administered with 30 mg / kg dose, and solvent control group was administered with equal volume of 0.5% sucrose ester solution. Continuous intragastric administration for 5 days, once a day, 1 ml of blood was collected from the eyeball 1 hour after the last administration (3.8% sodium citrate solution was anticoagulated at 1:9), placed at 4°C for 5 minutes to induce aggregation, and 300 μl of anticoagulated blood was drawn On a dry glass slide coated with collodion solution, about 2×2cm 2 ; The blood slices were incubated in a constant-temperature forced-air drying oven at 37°C for 8 minutes, washed with pH=7.4 PBS to remove red blood cells and white blood cells, and air-dried at room temperature. After stainin...

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Abstract

The process of setting up cell membrane solid phase chromatographic model for screening active platelet aggregation components from Chinese medicine includes the following steps: establishing the fingerprint of ginger oleoresin solution as the Chinese medicine to be screened, separating chicken platelet suspension, immobilizing the effective component of ginger oleoresin solution with the platelet cell membrane, eluting the uncombined components, dissociating the platelet immobilized the effective component of ginger oleoresin, and the HPLC analysis of the dissociated solution of platelet immobilized effective component. The present invention has rich material source, low cost and excellent application foreground.

Description

technical field [0001] The invention belongs to the field of screening of active components of traditional Chinese medicine and research on mechanism of action, and in particular relates to a method for establishing a cell membrane solid-phase chromatography model for screening active components of traditional Chinese medicine for anti-platelet aggregation (antithrombotic effect). Background technique [0002] The traditional research mode of active ingredients of traditional Chinese medicine, that is, the extraction, separation and purification of a single component of Chinese medicine and the efficacy verification mode, has a huge workload, a long research cycle, and cannot fully reflect the characteristics of multi-component, multi-target and integrated effects of Chinese medicine. , so that the research on the material basis of Chinese medicine efficacy has been progressing slowly. [0003] Cell Membrane Immobilized Chromatography (CMIC) is a new type of bio-affinity chr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N33/50
Inventor 聂红孟兰贞罗勇黄雪松
Owner JINAN UNIVERSITY
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