Method for regulating polarization state of macrophages

A technology of macrophages, polarization state, applied in the field of biomedicine

Pending Publication Date: 2020-12-04
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the existing technology, there are few methods for regulating macrophage polarization, and the effect of regulating macrophage polarization to inhibit the occurrence and development of inflammation stil

Method used

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  • Method for regulating polarization state of macrophages
  • Method for regulating polarization state of macrophages
  • Method for regulating polarization state of macrophages

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Isolation and cultivation of bone marrow-derived macrophages

[0050] The mice were killed by neck dislocation, and soaked in two 75% alcohol beakers in turn; the thigh was cut off with scissors, the femur and tibia were separated, and the skin and flesh were removed and placed in 10 mL RPMI1640 medium (containing 10% FBS); the femur and tibia were cut On the articular surface, expose the bone marrow cavity, flush the bone marrow cavity with a 10mL syringe (1mL needle), and blow off the cells with a 1mL pipette gun;

[0051] After the cells were filtered through a cell sieve, the cell suspension was collected into a 50 mL centrifuge tube; centrifuged at 1000 g for 8 min, the supernatant was discarded, and the cells were resuspended in 1640 medium containing 10 ng / mL macrophage colony-stimulating factor (MCSF), and inoculated on 6 After three days, a medium containing 10 ng / mL MCSF was added, and the mature bone marrow-derived macrophages (BMDM) obtained after ...

Embodiment 2

[0052] Example 2 Effects of acetylcholine and cholinergic receptor agonists on M1 macrophages

[0053] RAW264.7 or BMDM were inoculated in 6-well plates, and when the cell density reached about 70%, different concentrations of acetylcholine (0, 10, 50, 100, 200 μM) or cholinergic receptor agonist GTS-21 were added (0, 5, 10, 20, 50 μM) for 10 minutes;

[0054] M1 macrophages were induced by lipopolysaccharide LPS or IFNγ stimulation, and the cells were collected 24 hours later to analyze the expression of pro-inflammatory factors TNFα, IL-1β and IL-6.

[0055] The result is as figure 2 As shown, in LPS-induced RAW264.7 cells, they were treated with different concentrations of acetylcholine (Ach) and GTS-21 for 24 hours, and then the relative expressions of pro-inflammatory factors TNFα, IL-1β and IL-6 were analyzed. The results It was shown that the pro-inflammatory factors induced by LPS could be inhibited by acetylcholine and GTS-21 in a concentration-dependent manner. S...

Embodiment 3

[0056] Example 3 Effects of acetylcholine and cholinergic receptor agonists on M2 macrophages

[0057] RAW264.7 or BMDM were inoculated in 6-well plates, and when the cell density reached about 70%, acetylcholine or cholinergic receptor agonist GTS-21 was added to treat for 10 minutes;

[0058] M2 macrophages were induced by IL-4 stimulation, and the cells were collected 24 hours later to analyze the expressions of anti-inflammatory factors arginase 1 (Arg1), TGF-β, and IL-10.

[0059] The result is as image 3 As shown, in IL4-induced M2 macrophages RAW264.7 and primary bone marrow-derived macrophages, acetylcholine (Ach) and GTS-21 can significantly promote the differentiation of M2 macrophages, and promote the expression of related marker genes such as Arg1. , TGF and IL-10 expression.

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Abstract

The invention provides a method for regulating the polarization state of macrophages. The method comprises the step of activating cholinergic anti-inflammatory pathways of the macrophages by adoptingcholinergic anti-inflammatory pathway agonists and/or vagus nerve electrical stimulation signals. According to the method in the invention, the cholinergic anti-inflammatory pathway agonists are adopted to stimulate the macrophages in vitro; or the electrical stimulation signals are adopted to stimulate vagus nerves in vivo; therefore, the cholinergic anti-inflammatory pathways are activated; polarization of M1 type macrophages is inhibited; polarization of M2 type macrophages is promoted; a regulating effect on macrophage polarization types is achieved; and the method has the important significance in the aspects of specifically regulating the functions of the polarized macrophages and diseases caused by the polarized macrophages.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to a method for regulating the polarization state of macrophages, in particular to a method for regulating the polarization state of macrophages by activating cholinergic anti-inflammatory pathways and its role in delaying the occurrence of atherosclerotic inflammation application in development. Background technique [0002] Macrophages are formed by myeloid progenitor cells differentiated into monocytes, enter the blood, migrate to tissues and differentiate. According to the difference in activation mode and function, macrophages are divided into classical type (M1 type) and replacement type (M2 type). Under the stimulation of bacterial toxin LPS or Th1 cytokines (TNFα, IFNγ), macrophages are activated and polarized into M1 type, express and release pro-inflammatory factors and reactive oxygen species, have the functions of killing pathogens, clearing necrotic tissue and promoti...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/444A61P29/00A61P9/10A61P9/00A61P1/00A61P35/00A61P25/00A61P31/00A61P19/02A61N1/36
CPCA61K45/00A61K31/444A61P29/00A61P9/10A61P9/00A61P1/00A61P35/00A61P25/00A61P31/00A61P19/02A61N1/36053A61N1/36014
Inventor 钱政江李洪超杨海洋李翔
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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