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Method for screening glutathione reductase inhibitors based on fluorescence recovery of carbon dots

A reductase inhibitor, glutathione technology, applied in fluorescence/phosphorescence, material analysis through optical means, measurement devices, etc., can solve problems such as high cost, high biological toxicity, and unstable fluorescence

Inactive Publication Date: 2018-03-20
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the materials used have disadvantages such as high biological toxicity, unstable fluorescence, and high cost.

Method used

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  • Method for screening glutathione reductase inhibitors based on fluorescence recovery of carbon dots
  • Method for screening glutathione reductase inhibitors based on fluorescence recovery of carbon dots
  • Method for screening glutathione reductase inhibitors based on fluorescence recovery of carbon dots

Examples

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Embodiment Construction

[0041] The method for screening glutathione reductase inhibitors by fluorescence recovery of the present invention will be further described below through specific examples.

[0042] 10 kinds of drugs are selected, and their inhibition to glutathione reductase activity is determined by the method of the present invention and screened.

[0043] In 50 μL HEPES buffer solution (30 mM, pH=7.2), yellow carbon dot solution (50 μg mL -1 ) and Ag + Ionic solution (8.0 μM) was vortexed for 10 s, placed in a water bath, incubated at 25°C for 2 min, and the incubated product was obtained ——Carbon dots / Ag + Ionic solution (50 μg mL –1 / 8.0 μM);

[0044] In 30 μL HEPES buffer solution (30 mM, pH 7.2), glutathione reductase (0.5 U mL –1 ) and different drugs (50 mM) were vortexed for 15 s, and incubated in a water bath at 25°C for 10 min; then 10 μL NADPH solution (3.0 mM) and 10 μL oxidized glutathione solution (2.0 mM ), vortexed for 15 s, and incubated in a water bath at 25°C for ...

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Abstract

The invention discloses a method for screening glutathione reductase inhibitors based on fluorescence recovery of carbon dots. Yellow fluorescence carbon dots for Ag<+> ion specific quenching serve asa fluorescence probe; glutathione reductase can perform catalytic reduction on oxidized glutathione to produce reduced glutathione in the presence of a coenzyme NADPH (Nicotinamide Adenine Dinucleotide Phosphate), and the reduced glutathione can perform competitive binding with the Ag<+> ion so as to further recover fluorescence of the carbon dots; and the glutathione reductase treated and inactivated by the glutathione reductase inhibitor cannot perform catalytic reduction on the oxidized glutathione, therefore, the fluorescence of the carbon dots cannot be recovered. On this basis, screening of the glutathione reductase inhibitors can be realized by virtue of the fluorescence recovery situation. The method for screening glutathione reductase inhibitors disclosed by the invention has thecharacteristics of being high in sensitivity, excellent in selectivity, low in cost, low in toxicity, environment-friendly, high in stability and the like, and has significances for screening relateddrugs based on the glutathione reductase inhibitor.

Description

technical field [0001] The invention relates to a screening method for glutathione reductase inhibitors, in particular to a method for screening glutathione reductase inhibitors based on fluorescence recovery of carbon dots, which belongs to the field of biochemical analysis and drug screening. Background technique [0002] Malaria is one of the most common infectious diseases, causing illness and death in tropical and subtropical countries. The clinical cases caused by malaria are as high as hundreds of millions every year, resulting in tens of thousands of deaths, most of them are infants and young children. In recent years, due to the increasing resistance of Plasmodium parasites, the mortality rate has been increasing. The study found that an increase in reduced glutathione content in Plasmodium falciparum would enhance its resistance to antimalarial drugs, while a decrease in reduced glutathione content would restore the sensitivity of drug-resistant strains to antimal...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6486
Inventor 周雷千佳丽
Owner LANZHOU UNIVERSITY
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