Method for evaluating kidney toxicity of compounds through detecting contents of creatinine in zebra fish tissues

A kidney toxicity and compound technology, applied in the field of toxicology detection, can solve problems such as difficulty in obtaining blood or urine samples, small size of zebrafish embryos, etc., and achieve the effects of reducing experimental costs, reducing errors, and improving accuracy and reliability.

Active Publication Date: 2014-10-29
BIOLOGY INST OF SHANDONG ACAD OF SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

Zebrafish embryos are small in size, and it is difficult to obtain blood or urine samples. At present, the data on creat...

Method used

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  • Method for evaluating kidney toxicity of compounds through detecting contents of creatinine in zebra fish tissues
  • Method for evaluating kidney toxicity of compounds through detecting contents of creatinine in zebra fish tissues
  • Method for evaluating kidney toxicity of compounds through detecting contents of creatinine in zebra fish tissues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Nephrotoxicity of Different Concentrations of Aristolochic Acid A on Zebrafish Embryos (4dpf)

[0064] Reagent: Aristolochic acid A standard was purchased from China National Institutes for Food and Drug Control, dissolved in dimethyl sulfoxide to prepare a 5 mg / mL storage solution, stored at 4°C, and the longest storage time was 1 month. Dilute to desired concentration.

[0065] A standard 24-well cell culture plate was selected, and an appropriate amount of aristolochic acid A stock solution was added to each well, and diluted to 2 mL with culture water to obtain three concentration groups of 0.5 μg / mL, 2 μg / mL and 4 μg / mL. The control group was an aqueous culture solution containing 0.5‰DMSO, with a volume of 2 mL. Carefully blow and mix the solutions in all the sample wells with a pipette, randomly pick up 4-day-old zebrafish embryos, and transfer them into the above sample wells, with 15 embryos in each well, 4 wells in each group, and three parallel re...

Embodiment 2

[0081] Embodiment 2: the toxic effect of citrinin on zebrafish embryo (4dpf) kidney

[0082] Reagents: Citrinin standard product was purchased from China National Institute for the Control of Pharmaceutical and Biological Products. It was hydrolyzed with a culture solution containing 25% (V / V) ethanol to prepare a 10 mM stock solution, stored at 4°C, and diluted to the required concentration with culture water during experiments.

[0083]A standard 6-well cell culture plate was selected, and an appropriate amount of citrinin stock solution was added to each well, diluted with culture water to a concentration level of 25 μmol / L, and the final volume of each well was 6 mL. The control group was cultured aqueous solution containing equal concentration of ethanol, with a volume of 6 mL. Carefully blow and mix the solution in all sample wells with a pipette, randomly pick up 4-day-old zebrafish embryos, and transfer them into the above sample wells, with 60 embryos per well, and th...

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Abstract

The invention relates to a method for evaluating kidney toxicity of compounds through detecting the contents of creatinine in zebra fish tissues. The method is applied to kidney toxicity researches. In the researches that utilize a zebra fish embryo model to study the toxic effect of compounds on the kidney, the creatinine component in embryo tissues is extracted, then the creatinine content is measured through a LC-MS (liquid chromatography-mass spectrum) method, the creatinine content can be taken as a detection index for evaluating the embryo kidney functions, and thus a rapid and precise compound kidney toxicity evaluation system with strong specificity is built.

Description

technical field [0001] The invention relates to a method for evaluating the nephrotoxicity of a compound by detecting the creatinine content of zebrafish tissue, belonging to the field of toxicology detection. Background technique [0002] The kidney is an important organ of the body, which can excrete metabolites and certain wastes and poisons in the body, regulate the balance of water and electrolytes in the body, and ensure the stability of the body's internal environment. As the main excretory organ of the body, the kidney is extremely susceptible to various metabolic wastes and poisons in the body, and these substances often have potential renal toxicity. With the development of society, the number of metal, smelting, and chemical companies has increased, coupled with the abuse of medicines and various pesticides, environmental pollution has become increasingly serious, the chances of people being exposed to nephrotoxic substances have increased, and the incidence of to...

Claims

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Application Information

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IPC IPC(8): G01N30/88
Inventor 王雪刘可春韩利文陈锡强何秋霞彭维兵王希敏侯海荣张云孙晨楚杰
Owner BIOLOGY INST OF SHANDONG ACAD OF SCI
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