Method for regulating polarization of macrophages based on autophagy-exosome pathway and application

A macrophage and cell technology, applied in the field of biomedicine, can solve the problem of macrophage polarization that has not yet been seen

Pending Publication Date: 2021-11-19
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] So far, there have been no reports about regulating the polarization and recruitment of macrophages induced by influenza virus through cell autophagy and cell exocrine blocking methods, and ultimately controlling influenza virus infection and acute pneumonia

Method used

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  • Method for regulating polarization of macrophages based on autophagy-exosome pathway and application
  • Method for regulating polarization of macrophages based on autophagy-exosome pathway and application
  • Method for regulating polarization of macrophages based on autophagy-exosome pathway and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1PI3

[0054] Example 1 PI3K inhibitors and exosome inhibitors inhibit macrophage recruitment and cell aggregation

[0055] The autophagy inhibitor LY294002 (1 μM) and the exosome inhibitor GW4869 (1 μM) were added to the Ana-1 macrophages infected with the virus in the lower chamber of the Transwell system for cell transmembrane migration, and the effect on GFP in the upper chamber was observed. + Effects on macrophage recruitment (dosing intervention 24 hours post-infection), as figure 1 Immunofluorescence analysis shown shows that viral infection recruits more GFP +Macrophages; when using the autophagy inhibitor LY294002 and the exosome inhibitor GW4869, the results showed that both similarly greatly attenuated the response to GFP + The recruitment of macrophages suppresses the excessive inflammatory response induced by influenza virus.

Embodiment 2

[0056] Example 2 Effects of Pan-PI3K Inhibitors and PI3Kγ Inhibitors on Macrophage Polarization

[0057] In this example, the effects of different PI3K inhibitors on the polarization of macrophages infected with influenza virus were compared, such as figure 2 As shown, when the pan-PI3K inhibitor LY294002 and the PI3Kγ inhibitor AS605240 were used, the transcriptional level of iNOS was significantly inhibited, the level of anti-inflammatory IL-10 was up-regulated, and the replication of influenza virus was not significantly reduced. Reduced M1 polarization of macrophages, IL-1β transcription.

Embodiment 3PI3

[0058] Example 3 Effects of PI3K inhibitors on autophagy and exocrine

[0059] This experiment compared the effects of the PI3Kγ inhibitor AS605240 on the autophagy and exocrine behavior of macrophages under infection and non-infection conditions, such as image 3 As shown in the Western blot pictures, compared with the normal group, the ratio of LC3II / I increased after virus infection, indicating that cell autophagy was enhanced; when the specific PI3Kγ inhibitor AS was used, the protein ratio of LC3II / I was significantly reduced, indicating that AS inhibited the autophagy induced by influenza virus in a dose-dependent manner; at the same time, the results showed that after influenza virus infection, the protein levels of exosomal CD63 and CD81 were significantly lower than those in the normal group, and when AS was used to block autophagy When phagocytic LC3 is produced, CD63 and CD81 proteins show a dose-dependent increase, and the changes of autophagic protein LC3 and exoc...

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Abstract

The invention belongs to the technical field of biological medicine, and relates to a method for regulating polarization of macrophages, and in particular, relates to a method for regulating polarization of macrophages based on an autophagy-exosome pathway and an application. According to the method, the polarization of the macrophages can be regulated by compounds that inhibit the autophagy behavior and the behavior of extracellular secretion; through confirmation of experiments, the autophagy and recruitment of the macrophage M1 can be regulated through cell autophagy-cell exosome pathway blocking, the excessive M1 activation and recruitment of the macrophage induced by influenza virus are antagonized, and the effect of inhibiting influenza virus infection is achieved. Furthermore, the compound for inhibiting the cell autophagy behavior and the cell exosome behavior can be used for preparing drugs for treating influenza virus infection.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a method for regulating the polarization of macrophages, in particular to a method for regulating the polarization of macrophages based on an autophagy-exocrine pathway and its use. Phagocytosis inhibitors and exosome inhibitors regulate the polarization function of macrophages, and are further used in the preparation of drugs for the treatment of influenza virus infection. Background technique [0002] The prior art discloses that influenza virus belongs to the genus Orthomyxovirus, and is an enveloped virus comprising 8 negative-sense single-stranded RNAs (ssRNA), which are tightly surrounded by nucleoproteins, constituting the core of influenza virus. Genome. Studies have shown that there are glycoproteins attached to the surface of influenza virus, such as hemagglutinin (Hemagglutinin, HA) and neuraminidase (Neuraminidase, NA). According to the difference of HA and NA, inf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G01N33/68C12N5/0786A61K45/00A61K31/4706A61K31/5377A61P31/16
CPCG01N33/5044G01N33/68C12N5/0645A61K45/00A61K31/4706A61K31/5377A61P31/16C12N2500/30G01N2333/70503
Inventor 史训龙耿萍夏成杰朱海燕周伟黄海
Owner FUDAN UNIV
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