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Method for evaluating toxicity of myocardial injury inducing agent and efficacy of myocardial injury treating agent by using zebra fish

A technique for myocardial injury and zebrafish, which is applied in the field of drug evaluation and screening, and can solve problems such as research on zebrafish models that do not involve myocardial injury diseases

Active Publication Date: 2017-03-15
HANGZHOU HUANTE BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] It can be seen that the reports in the aforementioned existing literature do not involve research on zebrafish models of myocardial injury diseases; especially how to use zebrafish models to study myocardial injury diseases induced by chemical drugs, and to evaluate the toxicity or treatment of myocardial injury inducers. There is no literature report on the method of drug efficacy

Method used

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  • Method for evaluating toxicity of myocardial injury inducing agent and efficacy of myocardial injury treating agent by using zebra fish
  • Method for evaluating toxicity of myocardial injury inducing agent and efficacy of myocardial injury treating agent by using zebra fish
  • Method for evaluating toxicity of myocardial injury inducing agent and efficacy of myocardial injury treating agent by using zebra fish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example 1 Determining the Optimal Developmental Stage of Zebrafish

[0097] 1. Selection of zebrafish

[0098] Take 4 to 5 pairs of wild-type AB strain zebrafish parents to mate, according to Westerfield [22] method for hatching embryos. Zebrafish at 2dpf, 3dpf, and 4dpf were observed under a dissecting microscope, and normal zebrafish were picked and transferred to three 6-well plates, 30 in each well.

[0099] Note: ① "dpf" in the present invention=day post fertilization refers to the number of days after fertilization of zebrafish, such as "3dpf" refers to zebrafish five days after fertilization, the same below.

[0100] ② "Microwell plate" in the present invention includes but is not limited to the aforementioned 6-well plate, and others such as 12, 24, 48, 96 or 384-well plates are acceptable, the same below.

[0101] ③ The "number of zebrafish" in the present invention includes but is not limited to the aforementioned 30 zebrafish per well, and different number...

Embodiment 2

[0110] Example 2 Evaluating the Efficacy of Myocardial Injury Inducing Agent Isoproterenol

[0111] According to the optimal developmental stage of zebrafish determined in Example 1, a zebrafish myocardial injury model was established, and the efficacy of the myocardial injury inducer isoproterenol was evaluated.

[0112] 1. Selection of zebrafish

[0113] The zebrafish at 3 dpf determined in Example 1 were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred into 6-well plates, 30 in each well.

[0114] 2. Inducer treatment

[0115] Set up 6 experimental groups: 1 normal control group, 5 inducer treatment groups with different administration concentrations. In the normal control group, no treatment was given to the zebrafish; in the inducer treatment group, the zebrafish were treated with different concentrations of isoproterenol in water solution, the concentrations were 20mg / mL, 30mg / mL, 40mg / mL, 50mg / mL and 60mg / mL.

...

Embodiment 3

[0134] Example 3 Evaluating the Effect of Myocardial Injury Therapeutic Agent Digoxin

[0135] According to the optimal development stage of zebrafish determined in Example 1 and the optimal administration concentration of inducer determined in Example 2, a zebrafish myocardial injury model was established, and the therapeutic efficacy of digoxin, a therapeutic agent for myocardial injury, was evaluated.

[0136] 1. Selection of zebrafish

[0137] The zebrafish at 3 dpf determined in Example 1 were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred into 6-well plates, 30 in each well.

[0138] 2. Treatment of therapeutic agents

[0139] Set up 7 experimental groups: 1 normal control group, 1 model control group, and 5 therapeutic agent treatment groups with different dosage concentrations. In the normal control group, no treatment was carried out to the zebrafish; in the model control group, 50 mg / mL isoproterenol determine...

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Abstract

The invention establishes a myocardial injury model for a zebra fish, and provides a method for evaluating the toxicity of a myocardial injury inducing agent and the efficacy of a myocardial injury treating agent by using the zebra fish. The invention provides a brand-new medicine evaluating model and a brand-new medicine evaluating method; the medicine evaluating model and the medicine evaluating method have the advantages of reliability, fastness, high efficiency, cheapness, high cost performance and the like, can be used for evaluating the efficacy of disclosed medicines, can be also used for screening undisclosed medicines adopting brand-new compound structures, and are wide in application and high in significance.

Description

technical field [0001] The invention belongs to the technical field of drug evaluation and screening, and specifically relates to the application of a simple, economical and high-throughput zebrafish myocardial injury model, especially a method for evaluating the toxicity of myocardial injury-inducing agents and the efficacy of therapeutic agents using the zebrafish model . Background technique [0002] With the improvement of living standards and population aging, and affected by social and environmental factors, the incidence of cardiovascular disease is increasing year by year, and it has become one of the diseases with the highest mortality in human beings. In the occurrence and development of various cardiovascular diseases such as myocardial infarction and ischemia-reperfusion injury, myocardial injury is often accompanied by apoptosis, calcium overload, etc., causing cardiovascular dysfunction. According to the latest detection report of myocardial ischemia injury, d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00
CPCA61K49/0008
Inventor 朱晓宇夏波郭胜亚劳乔聪李春启
Owner HANGZHOU HUANTE BIOLOGICAL TECH CO LTD
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