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Application and detection method of dorsal aorta diameter in juvenile zebrafish as an effect marker in detecting PCB126 cardiovascular toxicity

A PCB126, dorsal aorta technology, applied in the field of toxicological chemical toxicity evaluation, can solve the problems of large initial sample size, high price, long life cycle, etc., and achieve the effect of protecting people's health and promoting social and economic development.

Active Publication Date: 2020-12-04
SUN YAT SEN UNIV
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  • Claims
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Problems solved by technology

Its main evaluation indicators are carcinogenicity, teratogenicity, endocrine disrupting effects, oxidative stress, etc.; the main mechanism is the induction and activation of aryl hydrocarbon receptor (AHR), which is affected by experimental models (long life cycle, time-consuming, expensive) and experimental techniques ( The inherent limitations of small throughput, large initial sample size, and low sensitivity cannot be used for rapid, accurate, reliable, and sensitive risk assessment under the conditions of acute exposure to PCB126

Method used

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  • Application and detection method of dorsal aorta diameter in juvenile zebrafish as an effect marker in detecting PCB126 cardiovascular toxicity
  • Application and detection method of dorsal aorta diameter in juvenile zebrafish as an effect marker in detecting PCB126 cardiovascular toxicity
  • Application and detection method of dorsal aorta diameter in juvenile zebrafish as an effect marker in detecting PCB126 cardiovascular toxicity

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Experimental program
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Effect test

Embodiment

[0083] Diameter measurement and statistical analysis of dorsal aorta in juvenile zebrafish:

[0084] 1. PCB126 exposure and fish egg culture:

[0085] The Tg(kdrl:eGFP) transgenic zebrafish eggs were exposed to PCB126 within 2 hours of fertilization, and the exposure dose was designed according to the literature reports and the previous research of our laboratory: 0, 0.0003, 0.3, 3, 30, 100, 300μg / L, cultivated to 72hpf in a light constant temperature incubator at 26.5°C, 14h light / 10h dark cycle. The poisoning solution was updated every 24 hours, and additional PTU (N-phenylthiourea) was added from 24hpf to inhibit pigmentation.

[0086] 2. Preparation of agarose-coated 96-well microplates:

[0087] Fill an appropriate amount of 2.5% hot agarose into a glass-bottom 96-well plate using a multichannel pipette. Insert 3D printed custom positioning molds (eg figure 1 shown), wait for the agarose to solidify (at least 15 minutes at room temperature), and carefully remove the p...

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Abstract

The invention discloses an application of a diameter of aorta dorsalis of a juvenile zebrafish as an effect marker in detection of PCB126 cardiovascular toxicity, and a detection method. The diameterof aorta dorsalis of the juvenile zebrafish is obtained through subjecting a roe of the juvenile zebrafish to PCB126 contamination treatment within 2 hours of fertilization, fixing a posture after culturing the roe, performing image acquisition and carrying out measurement. The juvenile zebrafish is a transgenic juvenile zebrafish of which vascular endothelial cells are labeled with green fluorescence. The juvenile zebrafish is cultured in an illumination constant-temperature incubator with the temperature of 26.5 DEG C in a 14h / 10h light and dark cycle, and is cultured to 72 hpf. The image acquisition is performed by adopting a high-content imaging system for layer scanning to obtain a juvenile zebrafish blood vessel fluorogram. The measurement is carried out by measuring the diameter ofthe aorta dorsalis by using Photoshop software, corresponding original data is recorded by means of EXCEL, and then software GraphPad Prism is used for statistical analysis. The application is beneficial to characterizing cardiovascular toxicity caused by low-dose PCB126 exposure at an ambient level, and provides a technical method and a reference basis for environmental monitoring and evaluationas well as formulation of related technical standards.

Description

technical field [0001] The invention relates to the application and detection method of an effect marker in detecting the toxicity of polychlorinated biphenyls, belonging to the technical field of toxicity evaluation of toxicological chemicals, in particular to the diameter of the dorsal aorta of zebrafish larvae as an effect marker in the detection of PCB126 cardiovascular Toxicity applications and detection methods. Background technique [0002] Polychlorinated Biphenyls (PCBs) are a family of artificially synthesized chlorinated aromatic hydrocarbon compounds consisting of 209 homologues, which are formed by replacing the hydrogen atoms of the biphenyl ring with 1-10 chlorines. Due to its good stability, insulation and heat resistance, PCBs have not only been widely used in transformers, capacitors, flame retardants, heat carriers and other industries, but also in many industrial resins, rubbers, plastics, coatings, etc. additives. However, because of its semi-volatile,...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6486
Inventor 韦艳宏钟霞丽吴景威柯炜健
Owner SUN YAT SEN UNIV
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