Anti-oxidation medicine screening kit and use method thereof

A kit and anti-oxidation technology, applied in the field of biomedicine, can solve the problems of nematode screening models without intellectual property protection and retention, and achieve the effects of intuitive and reliable results, simple use methods, and simple and fast operations

Inactive Publication Date: 2017-07-14
LANZHOU UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, the establishment of the above-mentioned high-throughput screening model for antioxidant drugs in C. elegans requires extremely high technology and experience. Even in developed countries, high-throughput drug screening technology for nematodes is still in the laboratory stage and has not yet entered industrialization. stage, and currently many nematode screening models do not have intellectual property protection

Method used

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  • Anti-oxidation medicine screening kit and use method thereof
  • Anti-oxidation medicine screening kit and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The assembly of embodiment 1 kit

[0063] A kind of antioxidant drug screening kit according to the present invention, the kit includes the following reagents:

[0064] ①. Reagent A: Caenorhabditis elegans suspension;

[0065] ②. Reagent B: nematode culture medium;

[0066] ③. Reagent C: Escherichia coli OP50;

[0067] ④. Reagent D: 2′,7′ dichlorofluorescein ethylene glycol;

[0068] The kit also contains a multiwell plate, which is black with Parafilm.

[0069] The porous plate is a 384-well microplate.

[0070] Wherein, the concentration of the reagent A is 50 Caenorhabditis elegans / 20 μl, the concentration of the reagent C is 10 mg Escherichia coli / ml, and the volume of the reagent B is 25 ml.

[0071] Wherein, the reagent C is a freeze-dried powder or a bacterial solution containing live Escherichia coli OP50.

[0072] The reagent B is S liquid culture medium.

[0073] Divide the above reagents and multi-well plates into suitable containers and place them in ...

Embodiment 2

[0076] Example 2 Synchronization of Caenorhabditis elegans

[0077] The Caenorhabditis elegans in the reagent A of the present invention is a nematode hatched to the L1 stage after synchronization, and the steps of Caenorhabditis elegans synchronization are as follows: wash down the Caenorhabditis elegans on the plate with M9 buffer, Inhale the washing solution into a centrifuge tube; centrifuge at 4000rpm for 3min, remove the supernatant; add the lysate; shake on a vortex instrument for 7min to rupture the worm body of Caenorhabditis elegans and release eggs; centrifuge at 4000rpm for 3min, remove the supernatant, and collect the precipitate to obtain A large number of Caenorhabditis elegans eggs; washed twice with M9 buffer, then added 1ml of M9 buffer, placed in an incubator at 20°C for 48 hours to incubate to obtain Caenorhabditis elegans hatched to the L1 stage after synchronization.

Embodiment 3

[0078] The use of embodiment 3 kit

[0079] (1) Use the Antioxidant Drug Screening Kit to screen antioxidant drugs, set a blank control group and a drug solution measurement group, and set each well in the multi-well plate as an independent experimental unit;

[0080] (2) Drug solution measurement group, add reagent B, the solution containing the drug solution to be tested, 5 μl reagent C and 20 μl reagent A to the wells of the multi-well plate in sequence under sterile conditions, so that the total volume of each well is 50 μl, set Set four parallel;

[0081] (3) Blank control group, under aseptic conditions, add reagent B, sterile water or vehicle equal to the solution of the drug solution to be tested, 5 μl reagent C and 20 μl reagent A to the wells of the multi-well plate in sequence, so that each well The total volume is 50 μl, and four parallels are set; wherein, the solution of the organic solvent is DMSO and ethanol;

[0082] (4) After adding the sample, seal the mul...

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Abstract

The present invention belongs to the technical field of biomedicine, and particularly relates to an anti-oxidation drug screening kit and a use method thereof. According to the present invention, the anti-oxidation drug screening kit comprises Caenorhabditis elegans, a nematode culture medium, Escherichia coli OP50, and 2',7' dichlorofluorescin diacetate, and can be used for high-throughput screening of anti-oxidation drugs; and kit has characteristics of simple operation and rapid detection, and is suitable for the large-scale promotion in the field of anti-oxidation drug screening.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a high-throughput screening kit for antioxidant drugs and a use method thereof. Background technique [0002] In recent years, with the rapid development of society, the fast-paced living environment makes people extremely vulnerable to oxidative stress damage caused by various stresses, and excessive oxidative stress damage is the cause of premature aging, diabetes and cardiovascular system diseases, etc. Factors that cannot be ignored in severe illness. Therefore, it is imminent to study drugs with low toxicity and significant antioxidant effect. [0003] At present, the commonly used in vivo animal models for antioxidant drug screening are mainly cells, mice, and rats. Mice and rats are ideal models for testing the efficacy of antioxidant drugs, but their application in the primary screening of a large number of antioxidant drugs is greatly limited by the pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/64
Inventor 李红玉王爱霞刘燕支德娟李洋
Owner LANZHOU UNIVERSITY
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