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Method for establishing hydrogen peroxide induced oxidation stress model of mouse monocyte macrophage system

A mononuclear macrophage and hydrogen peroxide technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., to achieve the effect of low cost and easy operation

Inactive Publication Date: 2017-01-04
GUANGXI UNIV
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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a method for establishing a mouse mononuclear macrophage cell line oxidative stress model induced by hydrogen peroxide with easy operation and low cost

Method used

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  • Method for establishing hydrogen peroxide induced oxidation stress model of mouse monocyte macrophage system
  • Method for establishing hydrogen peroxide induced oxidation stress model of mouse monocyte macrophage system

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Experimental program
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Embodiment Construction

[0013] 1. Experimental method

[0014] 1 Effect of different concentrations of hydrogen peroxide treatment on cell viability

[0015] (1) Experimental grouping and processing

[0016] According to the method reported by S Mueller et al., the OD was measured by UV spectrophotometer 240 After the value, press the formula: H 2 o 2 (mM)=OD 240 ×1000 / 39.4 calculation H 2 o 2 concentration.

[0017] The cell control group and hydrogen peroxide groups with different concentration gradients (25 μM, 50 μM, 100 μM, 150 μM, 200 μM, 250 μM, 300 μM, 350 μM, 400 μM, 450 μM) were respectively set up, and DMEM containing 5% FBS (or RPMI1640 containing 5% FBS ) medium to adjust the concentration of RAW264.7 cells to 4×10 5 cells / mL, and seeded in 96-well plate, 100 μL per well, 37°C, 5% CO 2 Cultivate overnight in the incubator; add 100 μL of different concentrations of hydrogen peroxide diluted with serum-free cell culture medium to each well of the experimental group, and add the sa...

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Abstract

The invention discloses a method for establishing a hydrogen peroxide induced oxidation stress model of a mouse monocyte macrophage system. The mouse monocyte macrophage system is treated by adopting hydrogen peroxide in vitro induction, and the concentration of hydrogen peroxide is 25 to 450 microns. Compared with the prior art, the mouse monocyte macrophage system is firstly used as treatment cells of the hydrogen peroxide induced oxidation stress model, the optimal using concentration and cell incubating time of an H2O2 solution are determined by inspecting a cell mortality rate and an intracellular ROS level, the method is easy to operate and relatively low in cost, and researchers for cultivation of mouse monocyte macrophage and an oxidation stress model firstly can be helped to complete the cultivation of the mouse monocyte macrophage and the establishment of the oxidation stress model relatively successfully.

Description

technical field [0001] The invention belongs to the technical field of mononuclear macrophage cell line oxidative stress models, in particular to a method for establishing a hydrogen peroxide-induced mouse mononuclear macrophage cell line (RAW264.7) oxidative stress model. Background technique [0002] Superoxide anion radicals are the earliest active oxygen radicals, and are the first products formed in the one-electron reduction reaction of oxygen. After being catalyzed by enzymes, superoxide anion can form other active oxygen radicals such as hydroxyl radicals, lipid peroxides, hydrogen peroxide and singlet oxygen. Its large-scale generation is an important reason for oxidative stress in the body. [0003] Hydroxyl free radical is the most toxic and harmful free radical to the body among active oxygen known so far. It can participate in various reactions of the body through electron transfer, addition and dehydrogenation, and interact with various macromolecules. , causi...

Claims

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Application Information

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IPC IPC(8): C12N5/0786
CPCC12N5/0645C12N2500/05
Inventor 胡庭俊郝祝兵李春节韦英益尹丹杨剑谭红连
Owner GUANGXI UNIV
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