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Method for screening drug action target position of tacrolimus in immunized T cell

A technology of tacrolimus and drug action, applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of large errors in screening results, huge analysis data, and undetermined drug action targets, etc., to achieve The results are true and reliable, and the operation ideas are clear

Inactive Publication Date: 2015-04-01
QINGDAO MUNICIPAL HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, clinically for individual patients, the drug target of tacrolimus in immune T cells has not been determined, and the conventional screening methods are too large and complex to analyze data, resulting in large errors in screening results and low accuracy, making it difficult to Provide test results with pharmacological significance and guidance during clinical medication

Method used

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Experimental program
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Effect test

Embodiment Construction

[0011] The immune T cells cultured to the fifth day were divided into the same amount of control group and several test groups, and different concentrations of tacrolimus were added to the test groups to form the overall effect of tacrolimus on the immune T cell model. drug effect sample group;

[0012] Step 2): Culture the drug effect sample group of tacrolimus on the immune T cell model described in step 1) to the logarithmic growth phase; discard the culture medium, scrape the cells and collect the cells into several corresponding centrifuge tubes; After the collection was completed, the cells were washed with phosphate buffer for 5 times, and the remaining phosphate buffer in the centrifuge tube was blotted after washing, and then an equal volume of 10mol / L urea, 50mmol / L DTT, 5% 3-[(3-cholamide propane base)-diethylamine 1-propanesulfonic acid, 30mmol / L trishydroxymethylaminomethane mixed solution, freeze and thaw cells repeatedly 5 to 8 times; put them in an ice-salt bat...

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Abstract

The invention relates to a method for utilizing a proteomic technology to screen a drug action target position of tacrolimus in an immunized T cell. The method for screening the drug action target position of tacrolimus in the immunized T cell is to utilize the proteomic technology to screen the drug action target position of tacrolimus in the immunized T cell and comprises the following steps: step 1) establishing sample groups of the action of tacrolimus immunizing the T cell; step 2) preparing two-dimensional electrophoresis protein samples of the immunized T cell; step 3) carrying out two-dimensional gel electrophoresis; step 4) collecting images and analyzing to obtain the result. The method for screening the drug action target position of tacrolimus in the immunized T cell, disclosed by the invention, is clear and definite in operation thought, and true and reliable in result, overcomes the defects that with adoption of the conventional screening method, the screening result is high in error and low in accuracy as the analysis data are huge and complicated, and provides the detection result with pharmacologic significance and guiding function for clinical medication.

Description

technical field [0001] The invention relates to a method for screening drug action targets of tacrolimus in immune T cells by proteomics technology. Background technique [0002] Tacrolimus, also known as FK506, is a fermentation product isolated from Streptomyces tsukubaensis, and its chemical structure is a 23-membered macrolide antibiotic. It is a powerful new type of immunosuppressant, mainly by inhibiting the release of interleukin-2 (L-2), and comprehensively inhibits the function of T lymphocytes, which is 100 times stronger than tacrolimus. In recent years, as a first-line drug for liver and kidney transplantation, it has been marketed in 14 countries including Japan and the United States. Clinical experiments have shown that it has good curative effect in heart, lung, intestine, bone marrow and other transplants. At the same time, FK506 also plays an active role in the treatment of autoimmune diseases such as atopic dermatitis (AD), systemic lupus erythematosus (S...

Claims

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Application Information

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IPC IPC(8): G01N27/447
Inventor 李化会莫晓媚刘丰海赵龙
Owner QINGDAO MUNICIPAL HOSPITAL
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