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

A technology of drug action and cyclosporine, applied in the direction of measuring devices, material analysis by electromagnetic means, instruments, etc., can solve the problems of huge analysis data, large error of screening results, difficulty in providing test results, etc., and achieve clear operation ideas , the result is real and reliable

Inactive Publication Date: 2015-04-01
QINGDAO MUNICIPAL HOSPITAL
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Problems solved by technology

At present, clinically for individual patients, the drug target of cyclosporine A in immune T cells has not been determined, and 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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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 cyclosporine A were added to the test groups to form the overall effect of cyclosporine A on the immune T cell model. drug effect sample group;

[0012] Step 2): Cultivate the drug action sample group of cyclosporine A 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 a...

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Abstract

The invention relates to a method for utilizing a proteomic technology to screen a drug action target position of cyclosporine A in an immunized T cell. According to the method for screening the drug action target position of cyclosporine A in the immunized T cell, the proteomic technology is used for screening the drug action target position of cyclosporine A in the immunized T cell and the method comprises following steps: step 1) establishing sample groups of the action of cyclosporine A 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 cyclosporine A in the immunized T cell, disclosed by the invention, is clear and definite in operation through, 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 the drug action target of cyclosporine A in immune T cells by proteomics technology. Background technique [0002] Cyclosporin A (cyclosprine, CsA) is a cyclic polypeptide composed of 11 amino acids, which is an active metabolite of a fungus in soil. In 1978, cyclosporine A was first used in clinical kidney transplantation in the United Kingdom. Since then, cyclosporine A has been used in transplantation of liver, heart, lung, pancreas, bone marrow and other organs. Satisfactory results have been achieved, and the survival of patients has been significantly improved. Rate. Cyclosporine A entered my country in 1984, and a triple immunosuppressive regimen of cyclosporine A, azathioprine and hormones was formed, which greatly improved the survival rate of transplantation, and also greatly reduced the incidence of acute rejection after transplantation. As a potent immunosuppressant, cyclosporin A has been s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62G01N27/447
Inventor 李化会莫晓媚刘丰海赵龙
Owner QINGDAO MUNICIPAL HOSPITAL
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