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Cell model for screening antiphlogistic medicament and method for screening medicament by using the same

A cell model, anti-inflammatory drug technology, applied in the direction of cells modified by introducing foreign genetic material, biochemical equipment and methods, microbial assay/examination, etc. characteristics and other issues, to achieve the effect of low cost, clear mechanism of action, and simple operation

Inactive Publication Date: 2009-06-24
TONGJI UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This screening method can realize the rapid screening of drugs, but because the immortalized cells cultured in vitro have lost some of the activities and characteristics of cells in the body, the screening results cannot fully reflect the effect of drugs in vivo

Method used

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  • Cell model for screening antiphlogistic medicament and method for screening medicament by using the same
  • Cell model for screening antiphlogistic medicament and method for screening medicament by using the same
  • Cell model for screening antiphlogistic medicament and method for screening medicament by using the same

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Experimental program
Comparison scheme
Effect test

Embodiment

[0050] Dexamethasone is a corticosteroid drug with pharmacological effects such as anti-inflammation, anti-endotoxin and enhancing stress response. In the inflammatory response, dexamethasone can regulate the release of inflammatory mediators and cytokines. We used IL-luc transgenic mouse macrophage inflammation model to evaluate the anti-inflammatory effect of dexamethasone.

[0051] 1. The isolated transgenic mouse macrophages were treated with 10 4 cells / well were seeded in a 96-well plate. After the cells were cultured for 60 h, dexamethasone was added to the cell culture medium to make the final concentration 1 μM. The cells were then placed at 37°C, 5% CO 2 Incubate in the incubator for 30 min.

[0052] 2. Add LPS or zymosan to the cell culture medium so that the final concentration is 100ng / ml or 125μg / ml respectively, and place at 37°C, 5% CO 2 Incubate for 3 h in the incubator.

[0053] 3. Aspirate the culture medium and wash the cells twice with PBS.

[0054] ...

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Abstract

The invention sets up an anti-inflammatory drug screening primary culture cell model which takes interleukin 1 beta gene of human beings as a target and is high-efficient and reliable, and a screening method thereof. The invention separates and primarily cultures a abdominal cavity macrophagus of the transgenic mouse which is introduced with interleukin 1 beta promoter drive of human beings and expressed by luciferase reporter gene, so that a macrophagus inflammatory model is set up by the inducement of inflammation motivating factor. In the model, the anti-inflammatory activity of the compound can be evaluated by measuring the degree of inhibiting the expression of the reporter gene of the screened compound. The drug screening model set up by the invention is a cell model which is sensitive, high-efficient, reliable and suitable for screening high flux anti-inflammatory drug.

Description

technical field [0001] The invention relates to a method for screening anti-inflammatory drugs. Background technique [0002] Inflammation is a pathological process in which living tissue responds mainly to defense against injury or infection. On the one hand, the inflammatory response is the body's defense response against injury factors, which is manifested in the increase of local blood supply, inflammatory cells entering the inflammatory site, and serum protein exudation, thereby controlling infection and removing harmful antigenic foreign bodies. However, inflammation is also a pathological process, and abnormal immune response may also lead to tissue necrosis, causing dysfunction, and even endangering the patient's life. Since inflammatory diseases have the characteristics of long course, easy recurrence, and difficult to cure, the development, evaluation and screening of anti-inflammatory drugs are of great practical significance for the treatment of inflammatory dis...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12Q1/66C12Q1/02
Inventor 荆华李利妹费俭王铸钢
Owner TONGJI UNIV
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