Method for adjusting polarization state of macrophage

A macrophage, polarization state technology, applied in the field of biomedicine, can solve the problem of few macrophages, and achieve the effect of promoting function

Inactive Publication Date: 2022-01-07
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are currently few effective m

Method used

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  • Method for adjusting polarization state of macrophage
  • Method for adjusting polarization state of macrophage
  • Method for adjusting polarization state of macrophage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Separation and culture of bone marrow deerived macrophage, BMDM

[0053] Due to the neck of mice, soak it in two 75% alcoholic alcohol; cut the thighs with scissors, separated the femur and tibia, and remove the flesh and placed in 10 ml RPMI1640 medium (containing 10% FBS); cutting the femur and the tibia The joint surface is exposed to the bone marrow chamber, and the bone marrow cavity is rinsed with 10ml syringe (1 ml needle), 1 ml of pipette blown cells;

[0054] After the cells were filtered through a cell sieve, the cell suspension was collected from the cell suspension to 50 ml of centrifuge tube; 1000 g from centrifugation for 8 minutes, discarded, and containing 1640 medium retransmitted by 10 ng / ml macrophage colony stimulating factor (MCSF), inoculated The 6-well plate was cultured; 3 days of medium containing 10 ng / ml MCSF were added, and the mature bone marrow source obtained after 3 days was cultured.

Embodiment 2

[0055] Example 2 Overgramps HIMF gene on M1 macrophages

[0056] Raw264.7 cells were seeded in 6-well plates, and when the cell density was about 70%, the adenovirus vectors of the virus titer Mo1-30 were added to the HIMF gene of the HIMF gene infected Raw264.7 cells. M1 macrophages were stimulated using lipopolysaccharide, LPS), and cells were collected after 48 hours, and the expression of associated marker gene TNFα, IL-1β, IL-6 was analyzed.

[0057] BMDM cells were inoculated in 6-well plates, and when the cell density was about 70%, BMDM cells were added to the viral titer MO1-30, respectively, and BMDM cells were infected. The expression of M1 macrophages was collected by IFN-γ stimulation, and the cells were collected after 48 hours, and the expression of the relevant marker gene TNFα, IL-1β, IL-6 was analyzed.

[0058] Such as Figure 3A and Figure 3b As shown, the results show that the HIMF gene can significantly promote the polarization of M1 macrophages compared to the...

Embodiment 3

[0059] Example 3 Effect of HIMF gene on M2 macrophages over expression

[0060] Raw264.7 cells were seeded in 6-well plates, and when the cell density was about 70%, the adenovirus vectors of the virus titer Mo1-30 were added to the HIMF gene of the HIMF gene infected Raw264.7 cells. M2 macrophages were induced using IL-4 stimulating, 48 hours of collection of cells, analyzed the expression of associated marker gene Arg1, TGF, and IL-10.

[0061] BMDM cells were inoculated in 6-well plates, and when the cell density was about 70%, BMDM cells were added to the viral titer MO1-30, respectively, and BMDM cells were infected. M2 macrophages were induced using IL-4 stimulating, 48 hours of collection of cells, analyzed the expression of associated marker gene Arg1, TGF, and IL-10.

[0062] Such as Figure 4A and Figure 4B As shown, the results showed that in IL-4-induced Raw264.7 cells, overexpression of HIMF genes can significantly inhibit the polarization of M2 macrophages, IL-4-induc...

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Abstract

The invention relates to the technical field of biological medicines and particularly discloses a method for adjusting a polarization state of a macrophage. The method comprises the step of adopting an over-expressed HIMF gene to infect the macrophage or adopting specific siRNA to knock down or integrally knock out an HIMF gene in the macrophage. Through the mode, the macrophage is infected by adopting the over-expressed HIMF gene or the HIMF gene in the macrophage is knocked down or integrally knocked out by adopting the specific siRNA, and functions of macrophages of different polarization types are influenced, so that the occurrence and development of inflammations are inhibited, and the occurrence of myocardial infarction is delayed.

Description

Technical field [0001] The present application relates to a biomedical field, and in particular, a method involving a state of regulating macrophages. Background technique [0002] Macrophages are polarized by bone marrow progenitor cells into monocytes into blood, migrate to polarization of the tissue. The macrophages are divided into a classic (M1) and alternative (M2) depending on the difference in activation and function. Under bacterial toxin LPS or TH1 cytokines (TNFα, IFNγ), macrophages are activated into M1, expression, and release related markers gene and active oxygen, with kill pathogens, remove necrotic tissue and promote inflammation. The effect of the reaction occurs; with the induction of TH2 cytokines (IL-4, IL-13), macrophages are activated as M2 type, express anti-inflammatory factors (such as TGFβ, IL-10), with Formulating inflammation, promoting the role of tissue growth and repair. [0003] The study found that two different polarized macrophages have importa...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/12C12N15/861C12N15/113A61K31/7088A61P29/00A61P9/00A61P31/04A61P19/02A61P1/00A61P25/00A61P35/00A61P9/10
CPCC12N5/0645C12N15/113C12N15/86C07K14/4702A61K31/7088A61P29/00A61P9/00A61P31/04A61P19/02A61P1/00A61P25/00A61P35/00A61P9/10C12N2510/00C12N2509/00C12N2509/10C12N2501/22C12N2710/10043
Inventor 钱政江李燕娇李翔李洪超杨海洋
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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