Method for rapidly evaluating damage effect on hepatic function of zebra fishes by compounds

A liver function and compound technology, applied in the field of toxicology detection, can solve the problems of large error in experimental results, large error in liver area selection, long experimental cycle, etc., achieve stable and reliable repeatability, improve experimental efficiency, and reduce experimental cost. Effect

Active Publication Date: 2015-12-02
BIOLOGY INST OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Chinese Patent Literature CN (Application No. 201010170675.2) discloses a method for screening liver injury drugs using the model organism zebrafish, but this method uses adult zebrafish for screening, and the experimental period is long, the cost is high, and it is not suitable for high-throughput screening.
[0006] Chinese patent document CN102353663A (application number 201110190076.1) discloses a method for quantitatively evaluating the hepatotoxicity of a compound with zebrafish. This method evaluates the hepatotoxicity of a compound by selecting the entire liver area of ​​the zebrafish and calculating the sum of the brightness of the liver. However, this Me

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  • Method for rapidly evaluating damage effect on hepatic function of zebra fishes by compounds
  • Method for rapidly evaluating damage effect on hepatic function of zebra fishes by compounds
  • Method for rapidly evaluating damage effect on hepatic function of zebra fishes by compounds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1: Evaluation of the toxic effect of carbaryl on zebrafish liver

[0065] 1. Acquisition and Use of Juvenile Zebrafish

[0066] Use healthy and sexually mature zebrafish whose liver specifically expresses fluorescence, put them in a mating tank at a ratio of 1 / 1 or 1 / 2, place a partition in the middle, and place it in a dark environment. Remove the partition before turning on the light the next day , stimulated by light to ovulate, fish out the adult fish half an hour later, and control the ovulation time within half an hour to reduce the difference in development time between embryos. Fertilized eggs were collected, sterilized and cleaned, then transferred into zebrafish embryo culture water, and 0.2ppm methylene blue was added to the culture water, and light-controlled culture was performed at 28°C. Add 0.2mM phenylthiourea 12 hours after the birth of the zebrafish embryos, change about 1 / 2 of the water every 24 hours in the middle, and suck out the dead embr...

Embodiment 2

[0094] Example 2: Evaluation of the toxic effect of isoniazid on zebrafish liver

[0095] 1. Acquisition and Use of Juvenile Zebrafish

[0096] Use healthy and sexually mature zebrafish whose liver specifically expresses fluorescence, put them in a mating tank at a ratio of 1 / 1 or 1 / 2, place a partition in the middle, and place it in a dark environment. Remove the partition before turning on the light the next day , stimulated by light to ovulate, fish out the adult fish half an hour later, and control the ovulation time within half an hour to reduce the difference in development time between embryos. Fertilized eggs were collected, sterilized and cleaned, then transferred into zebrafish embryo culture water, and 0.2ppm methylene blue was added to the culture water, and light-controlled culture was performed at 28°C. Add 0.2 mM phenylthiourea 12 hours after the birth of zebrafish embryos, change about 1 / 2 of the water every 24 hours, and suck out dead embryos in time. The 4-...

Embodiment 3

[0120] Example 3: Evaluation of the toxic effect of pyrazinamide on zebrafish liver

[0121] 1. Acquisition and Use of Juvenile Zebrafish

[0122] Use healthy and sexually mature zebrafish embryos whose liver specifically expresses fluorescence, put them in a mating tank at a ratio of 1 / 1 or 1 / 2, place a partition in the middle, and place it in a dark environment. Remove the partition before turning on the light the next day. Plates, stimulated by light to ovulate, fish out the adult fish half an hour later, and control the ovulation time within half an hour to reduce the difference in development time between embryos. Fertilized eggs were collected, sterilized and cleaned, then transferred into zebrafish embryo culture water, and 0.2ppm methylene blue was added to the culture water, and light-controlled culture was performed at 28°C. Add 0.2 mM phenylthiourea 12 hours after the birth of zebrafish embryos, change about 1 / 2 of the water every 24 hours, and suck out dead embryo...

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Abstract

The invention relates to a method for rapidly evaluating the damage effect on the hepatic function of zebra fishes by compounds. The method includes the steps that 1, fertilized normal zebra fish embryos are moved into breeding holes; 2, continuous chemical hatching is performed on the zebra fish embryos in the breeding holes, and the zebra fish embryos are marked to be a compound set to be detected; 3, livers of the zebra fish embryos are observed, and the liver area of the zebra fish embryos and the area of zebra fish embryo bodies are recorded and calculated; 4, according to the range relation between the index of the liver area of the zebra fish embryos of the compound set to be detected and the index of the liver area of normal zebra fish embryos, whether the compound set to be detected has liver toxicity is judged. The index of the liver area is used as a detection index for evaluating the hepatic function of the zebra fishes for the first time, and therefore the method is more scientific and objective and improves the accuracy of results. An established model for evaluating the damage effect on the hepatic function of the zebra fishes by the compounds has the advantages of being easy to manufacture, rapid to use, stable, reliable and good in repeatability.

Description

technical field [0001] The invention relates to a method for rapidly evaluating the effect of compounds on zebrafish liver function damage, and belongs to the technical field of toxicology detection. Background technique [0002] Drug toxicity is a key issue in the development of the pharmaceutical industry, and drug-induced liver injury is the most common adverse drug reaction. With the continuous advent of new drugs, the incidence of drug-induced liver injury has increased accordingly. The excipients of many medicines, Chinese herbal medicines and health care medicines may also cause liver damage. Drug-induced liver toxicity is one of the main reasons for the failure of new drug development or withdrawal from the market after marketing. According to statistics, about 1 / 3 of drugs are withdrawn from the market due to liver toxicity. Among the drugs approved by the FDA in the United States, more than 80% of the drugs with black box warnings belong to severe liver toxicity ...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 张云刘可春韩利文王雪何秋霞孙晨王希敏楚杰韩建王荣春
Owner BIOLOGY INST OF SHANDONG ACAD OF SCI
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