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Method for constructing oxidative stress model for porcine circovirus 2 (PCV2) in-vitro-infected porcine alveolar macrophage

A technology of alveolar macrophages and porcine circovirus, applied in the field of macrophage oxidative stress model, can solve the problems of immune cell redox state change, lack of understanding, etc.

Inactive Publication Date: 2016-07-27
GUANGXI UNIV
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Problems solved by technology

However, there is still a lack of knowledge about whether PCV2 infection can cause changes in the redox state of immune cells, and further research is urgently needed

Method used

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  • Method for constructing oxidative stress model for porcine circovirus 2 (PCV2) in-vitro-infected porcine alveolar macrophage
  • Method for constructing oxidative stress model for porcine circovirus 2 (PCV2) in-vitro-infected porcine alveolar macrophage
  • Method for constructing oxidative stress model for porcine circovirus 2 (PCV2) in-vitro-infected porcine alveolar macrophage

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Embodiment Construction

[0028] 1. Research ideas

[0029] The porcine alveolar macrophage cell line (3D4 / 2 cells) was infected by PCV2, and the level of nitric oxide (NO) secreted by the cells after virus infection, the level of total reactive oxygen species (ROS) in the cells, and the reduced glutathione (GSH) were measured. Content, xanthine oxidase (XOD) activity, myeloperoxidase (MPO) activity, inducible nitric oxide synthase (iNOS) activity, to explore the relationship between PCV2 virus infection amount, infection time and dynamic changes of reactive oxygen species level and establish an in vitro model of oxidative stress in porcine alveolar macrophages.

[0030] 2. Experimental method

[0031] (1) Cultivation of 3D4 / 2 cells: After recovery, 3D4 / 2 cells were transferred to bottles with RPMI1640 culture medium containing 10% fetal bovine serum, and incubated at 37°C and 5% CO 2 cultured in an incubator. Subculture when the cells grow to 70%-80%, digest with 0.05% trypsin and then subculture a...

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Abstract

The invention discloses a method for constructing an oxidative stress model for a porcine circovirus 2 (PCV2) in-vitro-infected porcine alveolar macrophage. The method comprises the steps of arranging a cell control group and a PCV2 infected group, separately adding an RPMI 1640 nutrient solution and a viral solution of corresponding concentration into each group, carrying out cell adsorption for 2 hours, then, discarding the solutions, adding 1mL of the RPMI 1640 nutrient solution containing 5% of FBS to each group, continuing to carry out culture, separately collecting samples, and applying the samples to the determination on cell activity and cell redox state related indexes. The model has the characteristics of convenience in constructing, relatively low cost, high practicability and the like, a relatively ideal in-vitro model is provided for researching whether PCV2 infection can cause the change of a redox state of an immunocyte or not and an action mechanism of the PCV2 infection, and then, some new thinking may be provided for the treatment on PCV2 infection induced animal diseases.

Description

technical field [0001] The invention belongs to the technical field of macrophage oxidative stress models, in particular to a method for constructing porcine alveolar macrophage oxidative stress models infected with porcine circovirus type 2 in vitro. Background technique [0002] Porcine circovirus disease caused by porcine circovirus type 2 (PCV2) infection has become a major disease affecting the swine industry worldwide, causing huge economic losses. The main sites of PCV2 virus replication are monocytes-macrophages and antigen-presenting cells of the body. PCV2 virus nucleic acid could be detected, and the main manifestations of PCV2 infection were lymphocyte loss and monocyte infiltration. Therefore, it causes the destruction of the pig's immune system, and the immunity is suppressed. The main site of PCV2 proliferation is B lymphocytes, and B lymphocytes also induce apoptosis of lymphocytes after PCV2 infection, causing immunosuppression. [0003] The phagocytosis ...

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Application Information

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IPC IPC(8): C12N5/0786
CPCC12N5/0645C12N2500/84
Inventor 胡庭俊尹丹谭红连韦英益郝祝兵杨剑
Owner GUANGXI UNIV
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