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Method for assessing acute toxicity of medicament by using model animal zebrafish fries

An acute toxicity, zebrafish technology, applied in the field of medicine, can solve the problems of high experimental cost, long animal experiment period, limited animal source, etc., and achieve the effect of saving drug dosage, eliminating experimental procedures, and high-throughput operation.

Active Publication Date: 2013-09-18
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the current conventional animal experiments can provide highly comparable screening and evaluation results, the general animal experiment cycle is long and the cost is high, which is not suitable for high-throughput drug screening
In many countries in Europe and the United States, vertebrates (including mice) are subject to increasingly strict protection, animal sources are limited, approval procedures are complicated, and experimental costs are becoming more and more expensive, which seriously affects the process of new drug development and the safety assessment of environmentally toxic substances

Method used

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  • Method for assessing acute toxicity of medicament by using model animal zebrafish fries
  • Method for assessing acute toxicity of medicament by using model animal zebrafish fries
  • Method for assessing acute toxicity of medicament by using model animal zebrafish fries

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Study on the acute toxicity of triptolide with zebrafish fry as a model animal

[0022] Test drug: triptolide;

[0023] Source: sigma

[0024] Physical and chemical properties: white powder, content greater than 98%;

[0025] Preparation method: Weigh triptolide according to the required concentration and dissolve it in 100% DMSO, then dilute the triptolide DMSO test solution in zebrafish fry culture solution at a ratio of 0.1%.

[0026] Animal: Zebrafish, provided by the Institute of Model Animals, Nanjing University.

[0027] Reagents: Dimethyl sulfoxide (DMSO) was purchased from Sigma Company, and the fry culture medium was 0.2g / L Instant sea ​​salt. instant The sea salt was purchased from the local flower, bird, fish and insect market.

[0028] experimental method:

[0029] Take 70 zebrafish fry that were fertilized for 72 hours and develop normally, and randomly distribute them in a 48-well plate, with 10 in each well, and put them in 1ml of culture mediu...

Embodiment 2

[0034] Study on the Acute Toxicity of Paclitaxel Using Zebrafish Fry as a Model Animal

[0035] Test drug: paclitaxel;

[0036] Source: sigma

[0037] Physical and chemical properties: white powder, content greater than 98%;

[0038] Preparation method: Weigh paclitaxel according to the required concentration and dissolve it in 100% DMSO, and then dilute the paclitaxel DMSO test solution in the zebrafish fry culture medium at a ratio of 0.1%.

[0039] Animal: Zebrafish, provided by the Institute of Model Animals, Nanjing University.

[0040] Reagents: Dimethyl sulfoxide (DMSO) was purchased from Sigma Company, and the fry culture medium was 0.2g / L Instant seasalt. instant The sea salt was purchased from the local flower, bird, fish and insect market.

[0041] experimental method:

[0042] Take 90 zebrafish fry that were fertilized for 72 hours and develop normally, and randomly distribute them in a 48-well plate, with 10 in each well, and put them in 1ml of culture m...

Embodiment 3

[0047] Study on the Acute Toxicity of Paclitaxel Using Zebrafish Fry as a Model Animal

[0048] Test drug: fumagillin;

[0049] Source: sigma

[0050] Physical and chemical properties: white powder, content greater than 98%;

[0051]Preparation method: Weigh fumagillin according to the required concentration and dissolve it in 100% DMSO, and then dilute the fumagillin DMSO test solution in the zebrafish fry culture solution at a ratio of 0.1%.

[0052] Animal: Zebrafish, provided by the Institute of Model Animals, Nanjing University.

[0053] Reagents: Dimethyl sulfoxide (DMSO) was purchased from Sigma Company, and the fry culture medium was 0.2g / L Instant sea ​​salt. instant The sea salt was purchased from the local flower, bird, fish and insect market.

[0054] experimental method:

[0055] Take 70 zebrafish fry that were fertilized for 72 hours and develop normally, and randomly distribute them in a 48-well plate, with 10 in each well, and put them in 1ml of cultu...

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Abstract

The invention aims to provide a method for assessing the acute toxicity of a medicament by using model animal zebrafish fries. The method is easy to operate, rapid and high in accuracy. The method is characterized by comprising the following steps of preparing the medicament into solutions to be tested with various concentrations by using dimethyl sulfoxide or ethyl alcohol, selecting the zebrafish fries normal in development 72 hours after fertilization, randomly distributing the zebrafish fries into perforated plates, adding the medicament to be tested with various concentrations, observing under a microscope 96 hours after the fertilization, judging that the fries are dead after the hearts of the fries stop beating, counting the survival rate of each group of the zebrafish fries and carrying out statistics, counting the half lethal dose (LD50) value through fitting curve by using GraphPad Prism software and responding the acute toxicity of the medicament by using the LD50 value. The method has the advantages that the method is simple in operation, rapid and high in accuracy and the acute toxicity of the medicament to be tested can be judged preliminarily.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for evaluating the acute toxicity of medicines by using model animal zebrafish fry. Background technique [0002] At present, the internationally recognized experimental methods for drug screening and toxicity and safety evaluation are divided into in vitro (in vitro) and in vivo (in vivo) experiments. In vitro experiments are usually cell culture or biochemical experiments, and the efficacy or toxicity of drugs is judged by evaluating the effects of compounds on cells, biochemical enzymatic reactions, or signal transduction pathways. In vitro experiments have the advantage of being fast and efficient, but in vitro experiments usually do not involve the absorption, distribution, metabolism, and excretion of compounds, and the screening results are often inconsistent with the results of animal and human experiments, and the comparability is poor. In vivo exp...

Claims

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Application Information

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IPC IPC(8): A61K49/00
Inventor 何明芳高晓平佘金雄
Owner NANJING UNIV OF TECH
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