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A method for screening mizoribine drug action target in immune T cells

A drug action and mizoribine technology, which is applied in the field of screening mizoribine in the field of drug action targets in immune T cells, can solve the problems of huge analysis data, large errors in screening results, and low accuracy, and achieve clear operation ideas , the result is real and reliable

Inactive Publication Date: 2016-09-28
QINGDAO MUNICIPAL HOSPITAL
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Problems solved by technology

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

Method used

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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 mizoribine were added to the test groups to form the drug effect of mizoribine on the immune T cell model as a whole. sample group;

[0012] Step 2): Cultivate the drug effect sample group of mizoribine 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; collect After completion, rinse the cells with phosphate buffer 5 times, blot the remaining phosphate buffer in the centrifuge tube after washing, and then add an equal volume of 10mol / L urea, 50mmol / L DTT, 5% 3-[(3-cholamidopropyl )-diethylamine 1-propanesulfonic acid, 30mmol / L trishydroxymethylaminomethane mixed solution, freeze and thaw the cells repeatedly 5-8 times; put them in an ice-salt bath and let stand for 30min; then he...

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Abstract

The invention relates to a method for utilizing a proteomic technology to screen a drug action target position of mizoribine in an immunized T cell. According to the method for screening the drug action target position of mizoribine in the immunized T cell, the proteomic technology is used for screening the drug action target position of mizoribine in the immunized T cell and the method comprises the following steps: step 1) establishing sample groups of the action of mizoribine 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 mizoribine 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 mizoribine in immune T cells by proteomics technology. Background technique [0002] Mizoribine (Mizoribine, MZ, bredinin) Mizoribine is an imidazole antibiotic obtained by Asahi Kasei from the culture filtrate of soil mold. As an immunosuppressant, it has been used in clinical kidney transplantation in Japan since December 1991. Many clinical transplant centers in Japan have used mizoribine as a routine immunosuppressive drug after kidney transplantation. In recent years, it has also been used clinically as an anti-rejection drug for kidney transplantation in my country. Compared with similar drug azathioprine, mizoribine has less hepatotoxicity and bone marrow suppression. Its main adverse reactions are gastrointestinal reactions, blood system disorders and allergic symptoms, and occasionally bone marrow function suppression and acute renal failure. . At present, clinically...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/561
Inventor 李化会莫晓媚刘丰海赵龙
Owner QINGDAO MUNICIPAL HOSPITAL
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