Method for screening mitochondria targeted compounds by using zebra fish

Abstract

The invention relates to the field of compound detection, in particular to a method for screening mitochondria targeted compounds by using zebra fish. The method mainly comprises the following steps of: zebra fish selection, compound treatment, dyeing treatment and quantitative / qualitative analysis, wherein the dyeing treatment is that the dyeing treatment is performed on the zebra fish, on whichthe compound treatment is performed, for 0.5 to 1 hour at the temperature of 28 DEG C by taking JC-1 or DASPEI as a staining agent. The method covers the shortage that mitochondrial damage cannot be detected in vivo in the prior art by using an in vivo experimental analysis method for screening the mitochondria targeted compounds by using the zebra fish, is convenient, swift, economic and practical, and has the characteristics of high flux and high specificity.

Description

technical field [0001] The invention relates to the field of compound (chemical) detection, in particular to a simple, economical and high-throughput method for screening mitochondrial targeting compounds by using zebrafish. Background technique [0002] Mitochondria are important organelles in eukaryotic cells and participate in many important physiological and biochemical processes such as intracellular tricarboxylic acid cycle, fatty acid metabolism, and oxidative phosphorylation. Mitochondria play an important pivotal role in cell apoptosis. A series of metabolic processes in mitochondria are closely related to cell apoptosis, such as excessive production of oxygen free radicals, release of cytochrome C, and abnormal opening of mitochondrial membrane permeability transition pores. wait. The mitochondrial membrane has a variety of ion channels that can mediate ion transport, and the regulation of ion channels may affect the function of mitochondria and even cells. Mitoc...

AI Technical Summary

Problems solved by technology

The experimental operation of this method is complicated, it is easy to cause cell damage, and it cannot reflect the absorption, distribution, metabolism and excretion of the drug in the whole organism, and cannot reflect the real situation of the drug in the body.

[0011] (3) Cell transfection: transfect cells with cDNA encoding membrane protein labeled with green fluorescent protein (GFP), although this method can Applied to living cells and dynamic observation, but the process is more complicated and the processing capacity is low

[0016] (1) The detection at the cell level adopts the method of cell culture. Although this method is effective and requires low cost, the in vitro cells lack the metabolic cycle transformation of the whole organism and circulating distribution in the body, which cannot truly reflect the overall biological activity;

[0017] (2) The detection at the molecular level 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 drug's in vivo The true situation;

[0018] (3) Mitochondria will be damaged to varying degrees during the separation process, and the relative purity is relatively low;

[0019] (4) The enzyme activity of the isolated mitochondria is low, and the biological function is poor;

[0020] (5) It is difficult to avoid non-specific effects, resulting in false positive or false negative results;

[0021] (6) The sample volume is low, high-throughput screening cannot be performed, and quantitative analysis is difficult;

[0022] (7) The experiment has poor repeatability, poor specificity and poor sensitivity;

Although these methods are effective, the experimental operation is complicated, the experimental cost is high, and it is difficult to avoid non-specific effects, resulting in false positive or false negative results. The most important thing is that the cells in vitro lack the metabolic cycle transformation of the whole organism and the circulation distribution in the body. Does not truly reflect overall biological activity

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