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A method for measuring the effects of short-chain chlorinated paraffins on cellular energy metabolism pathways

A short-chain chlorinated paraffin and energy metabolism technology, applied in the field of molecular biology-based evaluation, can solve problems such as metabolic pathway research on energy metabolism, and achieve good repeatability, reduce differences, and small sample size

Active Publication Date: 2017-10-27
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

But so far most of the research is based on the changes of ATP / ADP, and few can study energy metabolism through metabolic pathways

Method used

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  • A method for measuring the effects of short-chain chlorinated paraffins on cellular energy metabolism pathways
  • A method for measuring the effects of short-chain chlorinated paraffins on cellular energy metabolism pathways

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0025] Embodiment: short-chain chlorinated paraffin (C 13 -CPs) exposure on cellular energy metabolism

[0026] Using HepG2 as a model cell, first culture and expose HepG2 cells, the steps are as follows:

[0027] HepG2 cells were seeded on six-well plates at a seeding density of 1.2×10 5 cells / mL, the volume of culture medium in each well of the six-well plate was 2.5 mL. Cultivate in the incubator for 24h (37°C, 5% CO 2 ), and when the cells were covered with about 80% of the area covered by the culture medium on the six-well plate, they were divided into four dose groups and exposed to short-chain chlorinated paraffin (C 13 -CPs, 55% Cl). That is, the original culture solution was discarded, and 2.5 mL of culture solution containing DMSO volume percentage of 0.05% and short-chain chlorinated paraffin concentrations of 0, 1, 10, and 100 μg / L were added to each well, and cultured for 24 hours (37 ° C, 5 %CO 2 ), the cell samples were collected, and the number of paralle...

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Abstract

The invention discloses a method for evaluating the influence of short-chain chlorinated paraffins on cell energy metabolism. Firstly, by exposing HepG2 cells to short-chained chlorinated paraffins, LC-MS is used to effectively analyze intracellular small molecule metabolites. Enzyme activity can be rapidly analyzed using biological detection methods. The changes of small molecule metabolites related to energy metabolism and the activity changes of rate-limiting enzymes that play a key role in energy metabolism were studied, and a method for evaluating energy metabolism toxicity of cells exposed to external stimuli or pollution was constructed based on molecular biology. The method can intuitively analyze the influence of pollutants on the energy metabolism pathway of cells, and the invention has the advantages of simple pretreatment method, fast detection, good repeatability, comprehensive information and the like.

Description

technical field [0001] The invention belongs to the field of pollutant toxicity evaluation, and relates to an evaluation method based on molecular biology, which evaluates the cytotoxicity of small-dose environmental pollutants through the change of pollutants on cell energy metabolism pathways. Background technique [0002] Short-chain chlorinated paraffins (Short-chain chlorinated paraffins, SCCPs) are chlorinated paraffins with a chain length of 10-13 carbon atoms and a chlorination degree of 30%-70% (by weight). The ability to migrate away from the environment, environmental persistence, bioaccumulation and other characteristics of POPs. Over the past 30 years, the United States, the European Union and Japan have conducted preliminary assessments of the ecological and human health risks of SCCPs. Existing toxicity studies have shown that: SCCPs are more toxic to aquatic animals; they have chronic toxic effects on invertebrates and fish; in carcinogenicity studies, it wa...

Claims

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Application Information

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IPC IPC(8): C12Q1/32
Inventor 耿柠波陈吉平王菲迪张海军孙晓丽
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI