Method for screening mitochondria targeted compounds by using zebra fish
A mitochondria and compound technology, applied in the field of compound (chemical) detection, can solve the problems of cell damage, difficult quantitative analysis, complicated experimental operation, etc., and achieve the effects of short sexual maturation cycle, avoidance of interaction, and simple administration method.
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Embodiment 1
[0180] Example 1 Targeting mitochondria to assess drug toxicity
[0181] The present embodiment carries out according to the following steps:
[0182] (1) Selection of zebrafish
[0183] The 4dpf zebrafish were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred into 48-well plates, 10 in each well.
[0184] (2) Compound treatment
[0185] Set up 8 experimental groups: 5 drug treatment groups to be tested, 1 mitochondrial damage positive control group, 1 solvent control group, and 1 blank control group. Remove the culture water in the microwell plate, and add 1mL doxorubicin with concentrations of 0.1μM, 1μM, 10μM, 100μM, and 1000μM to the five drug treatment groups to be tested respectively. [26] solution; in the positive control group, 1 mL of etoposide solution with a concentration of 50 μM was added; in the solvent control group, 1 mL of DMSO with a concentration of 0.1% was added; in the blank control group, 1 mL o...
Embodiment 2
[0202] Example 2 Targeting Mitochondria to Evaluate Environmentally Toxic Compounds
[0203] The present embodiment carries out according to the following steps:
[0204] (1) Selection of zebrafish
[0205] The 4dpf zebrafish were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred to a 48-well plate, with 10 fish per well.
[0206] (2) Compound treatment
[0207] Set up 8 experimental groups: 5 treatment groups of environmental toxic compounds to be tested, 1 mitochondrial damage positive control group, 1 solvent control group, and 1 blank control group. Remove the breeding water in the microwell plate, and add 1 mL of the environmentally toxic compound Nonglin at a concentration of 0.1 μM, 1 μM, 10 μM, 100 μM, and 1000 μM to the five treatment groups of environmentally toxic compounds to be tested. [27] solution; in the positive control group, 1 mL of staurosporine solution with a concentration of 50 μM was added; in ...
Embodiment 3
[0224] Example 3 Targeting mitochondria to screen anticancer drugs
[0225] Mitochondria can mediate apoptosis of tumor cells. Antineoplastic drugs act on mitochondria, leading to the opening of mitochondrial permeability transition pores, the decrease or disappearance of mitochondrial membrane potential, and then the decoupling of the respiratory chain, the depletion of glutathione, the generation of ROS and the release of cytochrome C and inducible factors, and induce tumors apoptosis. Therefore, mitochondrial toxic compounds have the potential to be used as potential anticancer agents.
[0226] The present embodiment carries out according to the following steps:
[0227] (1) Selection of zebrafish
[0228] The 4 dpf zebrafish were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred to a 96-well plate, with one fish per well.
[0229] (2) Compound treatment
[0230] Eight experimental groups were set up: 5 candidate...
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