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Method for establishing zebra fish thrombosis model

A technology of zebrafish and thrombus, which is applied in the field of establishment of zebrafish thrombus model, can solve the problems of lack of bio-integrated circulation conversion drug circulation distribution, high experimental cost, complicated experimental operation, etc.

Active Publication Date: 2013-02-06
南京新环检测科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This research is at the molecular level, and it also lacks the circulation conversion of the whole organism and the circulation distribution of drugs in the body, and the experimental operation is complicated, the experimental period is long, and the experimental cost is high, so it is not suitable for the screening of antithrombotic drugs

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  • Method for establishing zebra fish thrombosis model
  • Method for establishing zebra fish thrombosis model
  • Method for establishing zebra fish thrombosis model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0192] Example 1 Method for establishing a zebrafish thrombus model

[0193] Based on the optimal treatment stage of zebrafish and the optimal treatment time length of compounds, an in vivo zebrafish thrombus model was established by optimizing the concentration of phenylhydrazine. The design scheme is as follows:

[0194] 1 zebrafish selection

[0195] The 2dpf zebrafish were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred into 48-well plates, 10 in each well.

[0196] 2 compound treatment

[0197] Set up 7 experimental groups: 5 thrombus-inducing agent treatment groups, 1 solvent control group, and 1 blank control group. Remove the breeding water in the microwell plate, add 1mL of thrombus-inducing agent phenylhydrazine with concentrations of 10mM, 15mM, 20mM, 25mM, and 30mM to the thrombus-inducing agent treatment group; add 1mL of 0.1% alcohol to the solvent control group; Add 1mL of breeding water to it. Incuba...

Embodiment 2

[0211] Example 2 Evaluation of antithrombotic drugs using zebrafish thrombus model

[0212] 1 zebrafish selection

[0213] The 2dpf zebrafish were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred to a 12-well plate, 20 fish per well.

[0214] 2 compound treatment

[0215] Set up 8 experimental groups: 5 compound combination treatment groups, 1 thrombus model group, 1 positive control group, 1 solvent control group, and 1 blank control group. Remove the breeding water in the microwell plate, and add 2 mL of phenylhydrazine with a final concentration of 20 mM + sulfinpyrazone (antithrombotic drug) with a concentration of 0.1 μM, 1 μM, 10 μM, 100 μM, and 1000 μM to the compound combination treatment group; Add 2mL 20mM phenylhydrazine to the model group; add 2mL 60μM aspirin to the positive control group; add 2mL 0.1% alcohol to the solvent control group; add 2mL aquaculture water to the blank control group. They were c...

Embodiment 3

[0228] Example 3 Using the zebrafish thrombus model to screen antithrombotic drugs

[0229] 1 zebrafish selection

[0230] The 2 dpf zebrafish were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred to a 96-well plate, with one fish per well.

[0231] 2 compound treatment

[0232] Set up 9 experimental groups: 5 compound combination treatment groups, 1 thrombus model group, 1 positive control group, 1 solvent control group, and 1 blank control group. Remove the breeding water in the microwell plate, and add 150 μL of phenylhydrazine with a final concentration of 20 mM + dipyridamole (antithrombotic drug) with a concentration of 0.1 μM, 1 μM, 10 μM, 100 μM, and 1000 μM to the compound combination treatment group; Add 150 μL 20 mM phenylhydrazine to the thrombus model group; add 150 μL 60 μM aspirin to the positive control group; add 150 μL 0.1% alcohol to the solvent control group; add 150 μL culture water to the blank c...

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Abstract

The invention belongs to the field of drug screening, and specially relates to a method for establishing a zebra fish thrombosis model. The method comprises the following steps of 1) selecting a zebra fish, 2) carrying out compound treatment comprising the following processes of removing breeding water from micropore plates, respectively adding corresponding thrombosis inducer solution, a solventand breeding water into a thrombosis inducer treatment group, a solvent control group and a blank control group according to micropore plate sizes, and standing the micropore plate for culture at a constant temperature of 28 DEG C for 24 to 48 hours, wherein the thrombosis inducer solution is phenylhydrazine solution having concentration of 10 to 30 mM and the solvent is alcohol having concentration of 0.1%. and 3) carrying out microscope qualitative or / and quantitative analysis. The invention also provides a method for screening drugs through establishing the zebra fish thrombosis model. A zebra fish thrombosis model established through the method of the invention has the characteristics of simpleness, convenience, high efficiency and high flux.

Description

technical field [0001] The invention belongs to the field of drug screening (evaluation), and specifically relates to a simple, economical, fast, efficient, and high-throughput method for establishing a zebrafish thrombus model, and uses the animal model to screen antithrombotic drugs and evaluate the thrombogenicity of drugs . Background technique [0002] Thrombotic disease is a common cardiovascular and cerebrovascular disease caused by stenosis and occlusion of vascular lumen. Thrombosis is the main pathological basis of cardiovascular and cerebrovascular diseases that cause disability and high mortality such as stroke, coronary heart disease, and atherosclerosis. The initial stage of arterial thrombosis is mainly caused by damage to the vascular endothelium, platelet adhesion and aggregation, while venous thrombosis is generally caused by slow blood flow or stasis [1] . [0003] With the change of human living environment and dietary habits, the morbidity and mortali...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/15A61P7/02G01N21/64
Inventor 朱晓宇李春启
Owner 南京新环检测科技有限公司
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