Method for rapidly evaluating composition effect of estrogen and antiestrogen

An anti-estrogen and estrogen technology, applied in biochemical equipment and methods, material excitation analysis, microbial determination/inspection, etc. Observation and operation, the effect of many samples and large number of eggs laid

Inactive Publication Date: 2013-08-14
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the traditional fish chronic toxicity test is time-consuming and expensive, and cannot meet the requirements of rapid testing reflecting long-term effects

Method used

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  • Method for rapidly evaluating composition effect of estrogen and antiestrogen
  • Method for rapidly evaluating composition effect of estrogen and antiestrogen
  • Method for rapidly evaluating composition effect of estrogen and antiestrogen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 , Using zebrafish embryos to evaluate the compound effect of 17β-estradiol and tamoxifen

[0028] Test drug: 17β-estradiol (E 2 ), tamoxifen, all purchased from sigma company in the United States.

[0029] Preparation method: The standard product was dissolved in 0.1% acetone to prepare 1g / L mother liquor and stored at 4°C. The laboratory was diluted with ultrapure water to the required concentration.

[0030] Animal: Zebrafish.

[0031] Reagents: RT-PCR kit, purchased from Bao Biological Engineering (Dalian) Co., Ltd.

[0032] The specific steps of the test method are as follows:

[0033] 1. Obtaining zebrafish embryos: match 10 pairs of male and female fish at a ratio of 1:1 or 1:2, put them in a special zebrafish hatching box, separate the male and female fish with a partition, and place the broodstock under light after 12 hours , Start spawning within half an hour, suck out the eggs with a straw at the end of spawning, check for fertilization under a microscope, di...

Embodiment 2

[0047] Example 2 , Application of zebrafish embryos to evaluate the compound effect of o,p'-DDT and tamoxifen

[0048] Test drugs: o,p'-DDT, tamoxifen, all purchased from sigma company in the United States.

[0049] Preparation method: The standard product was dissolved in 0.1% acetone to prepare 1g / L mother liquor and stored at 4°C. The laboratory was diluted with ultrapure water to the required concentration.

[0050] Animal: Zebrafish.

[0051] Reagents: RT-PCR kit, purchased from Bao Biological Engineering (Dalian) Co., Ltd.

[0052] Test method: Compared with the test method of Example 1, the difference is that the selected mixture ratio of estrogen and anti-estrogens is: o,p'-DDT is 100μg / L, tamoxifen is 10μg / L, 50μg respectively / L and 250μg / L.

[0053] Test results: At the morphological level of embryonic development, o,p'-DDT and tamoxifen did not show any neutralizing or balancing effects, but a certain degree of toxic additive effects, such as Figure 5 , Image 6 with Fig...

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Abstract

The invention relates to a method for rapidly evaluating composition effect of estrogen and antiestrogen. The method comprises the following steps of: obtaining zebrafish embryos; mixing the estrogen with different concentration of antiestrogens to form different ratios of exposure liquid; putting the zebrafish embryos into the exposure liquid to expose for 96 hours lastingly, and maintaining the water temperature at 26+/-1 DEG C, wherein the photoperiod is 16 hours in daytime and 8 hours at night; exchanging 80% of exposure liquid after 24 hours, and recording the hatching rate, the aberration rate and the hatching time of the zebrafish embryos; collecting juvenile fishes to store for use after exposure is finished; and evaluating the composition effect on morphological level of embryonic development and/or gene transcription level. The method for evaluating the composition effect of estrogen and antiestrogen by the zebrafish embryos is simple, rapid and objective, just needs 96 hours, and is a novel method for rapidly evaluating the composition effect of the estrogen and the antiestrogen.

Description

Technical field [0001] The invention relates to the technical field of research on the combined toxicity effects of estrogen and anti-estrogens, in particular to a method for quickly evaluating the compound effects of estrogen and anti-estrogens. Background technique [0002] Environmental endocrine disruptors (EDCs) are widespread in the environment and pose a great threat to the ecological environment and human health. Toxicological studies have found that the compound effect of multiple EDCs is not a simple superposition of individual effects, but the result of the interaction of EDCs. In the real environment, multiple EDCs often exist at the same time, so it is especially necessary to study the compound effect of EDCs. In the current researches on compound effects, most of them are compound effects among similar EDCs. Research on compound effects among different types of EDCs is relatively lacking and urgently needs to be strengthened. Environmental estrogens and anti-estro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N5/073G01N21/64
Inventor 王鲁梅向霄钟玲盈吕晶夏兵
Owner SHANGHAI JIAO TONG UNIV
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