Method for rapidly evaluating potential teratogenicity of Chinese patent medicines by using embryo of zebrafish

A technology for zebrafish embryos and Chinese patent medicines, which can be used in the testing of pharmaceutical preparations, material inspection products, etc., can solve problems such as low practicability and no established criteria for potential teratogenicity.

Inactive Publication Date: 2017-10-03
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the past, there was no standard for judging the potential teratogenicity in the method of evaluating the developmental toxicity of traditional Chinese medicine by using zebrafish, especially there was no method for evaluating Chinese patent medicine, and the practicability was low

Method used

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  • Method for rapidly evaluating potential teratogenicity of Chinese patent medicines by using embryo of zebrafish

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] 1. Grind Fule granules into fine powder, accurately weigh 250 mg, add 250 microliters of DMSO, extract in an ultrasonic cleaner for 30 minutes, let stand at room temperature for 24 hours, and dilute to 50 milliliters with E3 culture. Centrifuge at 4000rpm, take the supernatant, the concentration of the supernatant is 5mg / ml. Dilute with E3 culture medium to obtain a series of drug solutions with concentrations of 0.08, 0.16, 0.31, 0.63, 1.25, 2.5 and 5 mg / ml.

[0012] 2. Place zebrafish embryos 3 hours after fertilization in a 96-well plate, place 1 embryo in each well, add 200 microliters of medicinal solution to each well, and culture 30 embryos with each concentration of medicinal solution. The orifice plate is placed in an environment of 28.5 degrees Celsius and a light / dark cycle of 12 / 12 (hours) for cultivation, and the medicinal solution is renewed every 24 hours until 72 hours after fertilization.

[0013] 3. Observe the morphology of zebrafish embryos at each ...

Embodiment 2

[0015] 1. Take a few Fuyanjing capsules, remove the capsules and take the drug core powder, accurately weigh 400 mg, add 250 microliters of DMSO, extract in an ultrasonic cleaner for 30 minutes, let it stand at room temperature for 24 hours, and culture and dilute with E3 to 50 ml, centrifuge at 4000 rpm for 20 minutes, and take the supernatant, the concentration of the supernatant is 8 mg / ml. The solution was diluted with E3 culture medium to obtain drug solutions with concentrations of 0.5, 1, 2, 4, 6 and 8 mg / ml.

[0016] 2. Place zebrafish embryos 3 hours after fertilization in a 96-well plate, place 1 embryo in each well, add 200 microliters of medicinal solution to each well, and culture 30 embryos with each concentration of medicinal solution. The orifice plate is placed in an environment of 28.5 degrees Celsius and a light / dark cycle of 12 / 12 (hours) for cultivation, and the medicinal solution is updated every 24 hours until 72 hours after fertilization.

[0017] 3. O...

Embodiment 3

[0019] 1. Cut Angong Niuhuang Pills into granules, accurately weigh 250 mg, add 250 microliters of DMSO, extract in an ultrasonic cleaner for 30 minutes, let stand at room temperature for 24 hours, and dilute to 50 milliliters with E3 culture. Centrifuge at 4000rpm, take the supernatant, the concentration of the supernatant is 5mg / ml. Dilute with E3 broth to give concentrations of 0.25, 0.38, 0.5, 0.75, 1, 1.5 and 2 mg / ml.

[0020] 2. Place zebrafish embryos 3 hours after fertilization in a 96-well plate, place 1 embryo in each well, add 200 microliters of medicinal solution to each well, and culture 30 embryos with each concentration of medicinal solution. The orifice plate is placed in an environment of 28.5 degrees Celsius and a light / dark cycle of 12 / 12 (hours) for cultivation, and the medicinal solution is updated every 24 hours until 72 hours after fertilization.

[0021] 3. Observe the morphology of zebrafish embryos at each concentration under a microscope and record ...

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Abstract

The invention relates to a method for rapid evaluation and testing of the potential teratogenicity of Chinese patent medicines. The method is characterized in that the embryo of zebrafish is taken as a toxicity model and exposed to an aqueous solution of a Chinese patent medicine; whether the embryo of zebrafish has developmental toxicity during development under the stimulation of the Chinese patent medicine is observed; and whether the Chinese patent medicine has potential teratogenicity is determined according to the indexes of toxicity of the Chinese patent medicine to the embryo of zebrafish. With the method, the toxicity of a plurality of frequently used gynecological Chinese patent medicines are successfully evaluated and the potential teratogenicity of the gynecological Chinese patent medicines are effectively tested, so significant guidance is provided for safe clinical usage of the Chinese patent medicines.

Description

technical field [0001] The invention belongs to the field of toxicity evaluation, in particular to a method for evaluating potential teratogenicity of Chinese patent medicines by using zebrafish embryos. Background technique [0002] Proprietary Chinese medicines are produced according to the prescriptions of traditional Chinese medicines using modern technology. During the production process, drug interactions different from traditional processing methods may occur, and new toxicity may occur. The traditional method of evaluating teratogenicity is to use the embryos of mammalian model organisms rats and mice for evaluation. However, due to the long development cycle of mammalian embryos and difficult observation, it is difficult to conduct rapid and large-scale evaluation. Generally, water-soluble components can be absorbed into the body in traditional Chinese medicine. The development of zebrafish embryos in aqueous solution is a new model suitable for the toxicity evaluat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/15
CPCG01N33/15
Inventor 姚美村陈洁烽陈缵光廖敏刘彤刘梦萍黄丹萍
Owner SUN YAT SEN UNIV
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