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Hyperuricemia cell model, building method thereof and application of model to uric acid reduction efficacy evaluation

A hyperuricemia and cell model technology, applied in the field of cell model construction, can solve the problems of long cycle and cumbersome drug screening experiments, and achieve the effect of short cycle and simple operation

Active Publication Date: 2016-12-07
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The primary purpose of the present invention is to propose a method for constructing a cell model of hyperuricemia for the efficient screening of drugs instead of animal models for the problems of cumbersome and long-term drug screening experiments in animal models of hyperuricemia

Method used

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  • Hyperuricemia cell model, building method thereof and application of model to uric acid reduction efficacy evaluation
  • Hyperuricemia cell model, building method thereof and application of model to uric acid reduction efficacy evaluation
  • Hyperuricemia cell model, building method thereof and application of model to uric acid reduction efficacy evaluation

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Embodiment 1

[0037] Construction of a hyperuricemia cell model: the experiment set up a blank control group and a model group.

[0038] Model group: HK-2 cells in the logarithmic growth phase were taken, digested with trypsin, blown with a pipette to make a suspension, and the cell concentration was adjusted to 10 by dilution. 5cells / mL, inoculated in a 12-well plate, cultured in a cell culture incubator for 12 h, discarded the cell supernatant, washed twice with PBS, added 0.25 mmol / L adenosine and incubated for 12 h, each well Add 0.1 IU xanthine oxidase and continue to incubate for 12 hours to obtain a hyperuricemia cell model;

[0039] Blank control group: no adenosine inducer was added, and other conditions were the same as the model group.

[0040] Take out the cell supernatant, analyze the cell supernatant by HPLC, the high performance liquid chromatogram and the uric acid content figure are respectively as follows image 3 and Figure 4 shown. Depend on image 3 It can be seen...

Embodiment 2

[0042] Construction of a hyperuricemia cell model:

[0043] Take the HK-2 cells in the logarithmic growth phase, add trypsin to digest them, blow them with a pipette to make a suspension, and adjust the cell concentration to 3×10 by diluting 5 cell / mL, inoculated in a 12-well plate, cultured in a cell culture incubator for 12 h, discarded the cell supernatant, washed twice with PBS, added 0.5 mmol / L adenosine and incubated for 12 h, each well Add 0.1 IU xanthine oxidase and continue to incubate for 12 hours to obtain a hyperuricemia cell model; take out the cell supernatant, and analyze the uric acid content in the cell supernatant by HPLC. The results are as follows: Figure 4 shown.

Embodiment 3

[0045] Construction of a hyperuricemia cell model:

[0046] Take the HK-2 cells in the logarithmic growth phase, add trypsin to digest them, blow them with a pipette to make a suspension, and adjust the cell concentration to 3×10 by diluting 5 cell / mL, inoculated in a 12-well plate, cultured in a cell culture incubator for 24 h, discarded the cell supernatant, washed 3 times with PBS, added 0.5 mmol / L adenosine and incubated for 24 h, each well Add 0.2 IU xanthine oxidase and continue to incubate for 24 hours to obtain a hyperuricemia cell model; take out the cell supernatant, and analyze the uric acid content in the cell supernatant by HPLC. The results are as follows: Figure 4 shown.

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Abstract

The invention relates to a hyperuricemia cell model, a building method thereof and an application of the model to uric acid reduction efficacy evaluation. The building method includes the steps: (1) taking human proximal tubular epithelial cells (HK-2 cells) as model making cells, dissociating the HK-2 cells in a logarithmic phase to prepare cell suspension, adjusting cell density and culturing the cells in a cell culture box for 12-36 hours after planking; (2) removing cell supernatant, cleaning the cells with PBS (phosphate buffer solution) for 2-3 times, adding adenosine and continuing incubation for 12-36 hours; (3) adding xanthine oxidase into the cell supernatant, continuing culture for 12-36 hours, taking out the cell supernatant, and performing efficient liquid detection of uric acid content. Experimental results indicate that the hyperuricemia cell model can be obtained by the aid of the inductor adenosine and the xanthine oxidase. The hyperuricemia cell model is built for the first time, uric acid reduction efficacy of a medicine can be simply and rapidly analyzed, and the model is of great significance for gout medicine research.

Description

technical field [0001] The invention belongs to the technical field of cell model construction, and in particular relates to a method for constructing a hyperuricemia cell model and its application in the evaluation of uric acid-lowering efficacy. Background technique [0002] Hyperuricemia is a metabolic disease caused by purine metabolism disorder and / or decreased uric acid excretion. It is mainly characterized by elevated blood uric acid and is an important biochemical basis for gout, kidney stones and other diseases. In recent years, the prevalence of hyperuricemia and gout has been increasing, and hyperuricemia, as one of many metabolic disorders, is often closely related to the development of hypertension, diabetes, obesity, atherosclerosis and other diseases , is an important disease threatening human beings. [0003] The current drugs for the treatment of hyperuricemia can be divided into: drugs that inhibit the production of uric acid; drugs that promote the excret...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12Q1/02
CPCC12N5/0686C12N2500/40C12N2501/71G01N33/5005
Inventor 任娇艳刘丹
Owner SOUTH CHINA UNIV OF TECH
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