Method for evaluating toxic effect mechanism

A technology of toxic effects and mechanisms, applied in biochemical equipment and methods, microbial determination/inspection, measurement devices, etc., can solve false positive signals, inability to evaluate mechanisms of toxic effects, and lack of standards for non-targeted metabolomics detection and analysis product verification, etc.

Inactive Publication Date: 2021-06-18
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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Problems solved by technology

[0003] At present, scholars at home and abroad generally use mice and zebrafish as research objects, and use non-targeted metabolomics to study the toxic effects of different types of EDCs at different times and concentrations. Standard verification, so many false positive signals may be generated, resulting in an inability to accurately evaluate the mechanism of toxic effects

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  • Method for evaluating toxic effect mechanism
  • Method for evaluating toxic effect mechanism
  • Method for evaluating toxic effect mechanism

Examples

Experimental program
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Embodiment 1

[0112] Using MCF-7 cells as the test biological sample, the combined toxic effects of two typical endocrine disruptors, PCB153 and atrazine, were studied based on metabolomics and transcriptomics analysis techniques.

[0113] Step (1): Cell culture and exposure

[0114] MCF-7 cells were incubated at 37°C, saturated humidity, 5% CO 2 Cultivate in a constant temperature incubator, and the medium is MEM liquid medium containing 10% fetal bovine serum, 100U / mL penicillin and 100U / mL streptomycin. MCF-7 cells grow adherently. Add 1mL trypsin solution to digest for 1min when subcultured. After the cells become round and gaps appear, add 2mL MEM liquid medium to stop the digestion. Repeated blowing to form a cell suspension, centrifuged at 1000r / min for 3min, After removing the supernatant, resuspend the cells in 2 mL of MEM liquid medium for passage.

[0115] Dilute the MCF-7 cell suspension in the logarithmic growth phase after 2-3 generations of recovery to 5.0×10 4 cells / mL,...

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Abstract

The invention discloses a method for evaluating a toxicity effect mechanism, and relates to the field of cytotoxicity evaluation methods. The method includes the following steps: after a biological sample is exposed to poison, screening out differential metabolites and differential genes through metabolomics detection and analysis and transcriptomics detection and analysis, and constructing a metabolic network composed of the differential metabolites and the differential genes through spearman correlation coefficient analysis to evaluate the toxic effect mechanism; the metabonomics detection and analysis comprises the following steps: performing non-targeted metabonomics detection and analysis to obtain differential metabolites of the biological sample after being exposed to the poison, and screening out a metabolic pathway having the highest correlation with the differential metabolites; and performing targeted metabonomics detection analysis on the basis of the non-targeted metabonomics differential metabolites to obtain the determined differential metabolites and related metabolic pathways.

Description

technical field [0001] The present application relates to the field of cytotoxicity evaluation methods, in particular, to a method for evaluating the mechanism of toxic effects. Background technique [0002] Endocrine Disrupting Chemicals (EDCs), because of their endocrine disrupting effects, have received widespread attention worldwide and have been identified as important risk factors affecting food safety. EDCs affect the internal environment stability, reproduction, development and even behavior of organisms by interfering with the synthesis, release, transport, binding, action and clearance of endogenous hormones. In recent years, there have been many food quality and safety incidents related to EDCs food-borne contamination at home and abroad. [0003] At present, scholars at home and abroad generally use mice and zebrafish as research objects, and use non-targeted metabolomics to study the toxic effects of different types of EDCs at different times and concentrations...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G16B5/00
CPCG01N33/5011G01N33/5014G01N33/5067G16B5/00
Inventor 许彦阳钱永忠邱静李耘
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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