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In-vivo test method for fluidity of mitochondrial membrane of liver of pseudorasbora parva

A test method and technology for mitochondrial membranes, applied in measurement devices, instruments, fluorescence/phosphorescence, etc., can solve problems such as poor biological function, membrane damage, and reduced mitochondrial activity in vitro, avoiding short-term effects and predictability. Good results

Active Publication Date: 2017-12-08
CHINESE RES ACAD OF ENVIRONMENTAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] (1) Although the in vitro mitochondrial membrane fluidity test is effective and requires low cost, the in vitro experiment lacks the metabolic cycle conversion of the whole organism and the circulation distribution in the body, and cannot truly reflect the overall biological activity;
[0006] (2) The in vitro detection needs to separate the mitochondria, which also lacks the absorption, distribution, metabolism and excretion process of the drug in the organism, and cannot reflect the real situation of the drug in the body;
[0007] (3) Mitochondria may destroy the natural state of the biofilm during the separation process, which will cause different degrees of membrane damage;
[0008] (4) The activity of isolated mitochondria is reduced, and the biological function is poor

Method used

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Examples

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Effect test

Embodiment 1

[0049] A method for measuring the fluidity of mitochondrial membranes in the liver of wheat ear carp in vivo, characterized in that it comprises the following steps:

[0050] S1. Fish exposure experiment

[0051] The water body poisoning method was adopted in the exposure experiment of the wheat ear fish body: the wheat ear fish was domesticated for 14 days indoors, and the mortality rate was stabilized below 5%, and it was randomly exposed to the glass tank of the drug solution treatment group, and the drug solution was replaced with the same concentration of drug for 24 hours. In order to keep the concentration of the drug solution as constant as possible, the poison exposure was carried out; respectively, 4 μg / L triazophos, 20 μg / L triazophos, 14 μg / L fipronil and 71 μg / L fipronil were used for poisoning 12 days, and the solvent-treated larvae were used as a parallel control, and samples were taken at the end of the exposure time, and 5 fish were taken each time as a treate...

Embodiment 2

[0074] A method for measuring the fluidity of mitochondrial membranes in the liver of wheat ear carp in vivo, characterized in that it comprises the following steps:

[0075] S1. Fish exposure experiment

[0076] The water body poisoning method was adopted for the exposure experiment of the wheat ear fish body: the wheat ear fish was domesticated for 10 days indoors, and the mortality rate was stabilized below 5%, and it was randomly exposed to the glass tank of the drug solution treatment group, and the drug solution was replaced with the same concentration of drug for 12 hours. In order to keep the concentration of the drug solution as constant as possible, exposure was carried out; 8 μg / L triazophos, 30 μg / L triazophos, 10 μg / L fipronil and 50 μg / L fipronil were used for poisoning respectively. 12 days, and the solvent-treated larvae were used as a parallel control, and samples were taken at the end of the exposure time, and 5 fish were taken each time as a treated sample. ...

Embodiment 3

[0097] A method for measuring the fluidity of mitochondrial membranes in the liver of wheat ear carp in vivo, characterized in that it comprises the following steps:

[0098] S1. Fish exposure experiment

[0099] The water body poisoning method was adopted in the exposure experiment of the wheat ear fish body: the wheat ear fish was domesticated for 16 days indoors, and the mortality rate was stabilized below 5%, and it was randomly exposed to the glass tank of the drug solution treatment group, and the drug solution was replaced with the same concentration of drug for 20 hours. In order to keep the concentration of the drug solution as constant as possible, the poison exposure was carried out; respectively, 4 μg / L triazophos, 20 μg / L triazophos, 14 μg / L fipronil and 71 μg / L fipronil were used for poisoning 12 days, and the solvent-treated larvae were used as a parallel control, and samples were taken at the end of the exposure time, and 5 fish were taken each time as a treate...

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Abstract

The invention relates to testing of fluidity of fluidity of a mitochondrial membrane of the liver of fish, in particular to an in-vivo test method for the fluidity of the mitochondrial membrane of the liver of pseudorasbora parva. The method comprises steps as follows: S1, a fish body exposure experiment; S2, preparation of the mitochondrial membrane of the liver of the pseudorasbora parva; S3, preparation of a DPH solution; S4, marking of a DPH fluorescent probe of the mitochondrial membrane; S5, determination of the fluidity of the membrane; S6, determination of the content of protein in mitochondria. With the adoption of the in-vivo test method for the fluidity of the mitochondrial membrane of the liver of pseudorasbora parva, influence of an agent on the fluidity of the mitochondrial membrane of the fish is determined accurately and conveniently.

Description

technical field [0001] The invention relates to the test of the fluidity of the mitochondrial membrane of the fish liver, in particular to a method for testing the fluidity of the mitochondrial membrane of the wheat ear fish liver in vivo. Background technique [0002] In recent years, with the continuous development of modern biotechnology, the research methods of the toxicological mechanism of insecticides are also developing towards the molecular level. One of these aspects is the study of biofilm effects. The fluidity of biomembrane phospholipid bilayer is one of the basic characteristics of biomembrane. A large number of studies have shown that the proper fluidity of biofilm is a necessary condition for the normal function of biofilm. For example, material transport, energy conversion, cell recognition and differentiation, cellular immunity, hormone action, membrane-bound enzyme activity, etc., are all closely related to membrane fluidity. An appropriate degree of me...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6439
Inventor 李会仙吴丰昌朱元荣
Owner CHINESE RES ACAD OF ENVIRONMENTAL SCI
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