Method for establishing zebra fish thrombosis model
A technology of zebrafish and thrombus, which is applied in the field of establishment of zebrafish thrombus model, can solve the problems of lack of bio-integrated circulation conversion drug circulation distribution, high experimental cost, complicated experimental operation, etc.
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Embodiment 1
[0192] Example 1 Method for establishing a zebrafish thrombus model
[0193] Based on the optimal treatment stage of zebrafish and the optimal treatment time length of compounds, an in vivo zebrafish thrombus model was established by optimizing the concentration of phenylhydrazine. The design scheme is as follows:
[0194] 1 zebrafish selection
[0195] The 2dpf zebrafish were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred into 48-well plates, 10 in each well.
[0196] 2 compound treatment
[0197] Set up 7 experimental groups: 5 thrombus-inducing agent treatment groups, 1 solvent control group, and 1 blank control group. Remove the breeding water in the microwell plate, add 1mL of thrombus-inducing agent phenylhydrazine with concentrations of 10mM, 15mM, 20mM, 25mM, and 30mM to the thrombus-inducing agent treatment group; add 1mL of 0.1% alcohol to the solvent control group; Add 1mL of breeding water to it. Incuba...
Embodiment 2
[0211] Example 2 Evaluation of antithrombotic drugs using zebrafish thrombus model
[0212] 1 zebrafish selection
[0213] The 2dpf zebrafish were observed under a dissecting microscope, and the normally developed zebrafish were picked and transferred to a 12-well plate, 20 fish per well.
[0214] 2 compound treatment
[0215] Set up 8 experimental groups: 5 compound combination treatment groups, 1 thrombus model group, 1 positive control group, 1 solvent control group, and 1 blank control group. Remove the breeding water in the microwell plate, and add 2 mL of phenylhydrazine with a final concentration of 20 mM + sulfinpyrazone (antithrombotic drug) with a concentration of 0.1 μM, 1 μM, 10 μM, 100 μM, and 1000 μM to the compound combination treatment group; Add 2mL 20mM phenylhydrazine to the model group; add 2mL 60μM aspirin to the positive control group; add 2mL 0.1% alcohol to the solvent control group; add 2mL aquaculture water to the blank control group. They were c...
Embodiment 3
[0228] Example 3 Using the zebrafish thrombus model to screen antithrombotic drugs
[0229] 1 zebrafish selection
[0230] The 2 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.
[0231] 2 compound treatment
[0232] Set up 9 experimental groups: 5 compound combination treatment groups, 1 thrombus model group, 1 positive control group, 1 solvent control group, and 1 blank control group. Remove the breeding water in the microwell plate, and add 150 μL of phenylhydrazine with a final concentration of 20 mM + dipyridamole (antithrombotic drug) with a concentration of 0.1 μM, 1 μM, 10 μM, 100 μM, and 1000 μM to the compound combination treatment group; Add 150 μL 20 mM phenylhydrazine to the thrombus model group; add 150 μL 60 μM aspirin to the positive control group; add 150 μL 0.1% alcohol to the solvent control group; add 150 μL culture water to the blank c...
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