In vivo activation of macrophages
In vivo activation of macrophages
Macrophages are important natural immune cells in vivo, and in vitro detection of their function does not fully reflect their in vivo behavior, while detection of the actual effects of certain drugs often requires in vivo measurement of macrophage activity. As an example, the bacterial endotoxin LPS activates macrophages, LPS is a complex of lipids and polysaccharides, which is the main component of the cell wall of Gram-negative bacteria, and it is an endotoxin and an important group-specific antigen.LPS causes sepsis, septic shock, and even death by inducing the activation of monocyte-phagocyte system, and the release of a large number of pro-inflammatory cytokines, such as TNFα, IL-6, etc.TLR4 TLR4 is the main pattern recognition receptor for LPS recognition by the monocyte-phagocyte system.
Principle
The basic principle of macrophage activation in vivo is that LPS, a complex of lipids and polysaccharides, is a major component of the cell wall of Gram-negative bacteria and is an endotoxin and an important group-specific antigen.LPS leads to the development of sepsis, septic shock, and even death by inducing activation of the monocyte-phagocyte system, which results in the release of large amounts of various pro-inflammatory cytokines, such as TNFα, IL-6, etc.TLR4 is the major pattern recognition receptor for the recognition of LPS by the monocyte-phagocyte system. TLR4 is the main pattern recognition receptor of monocyte-phagocyte system to recognize LPS.
Operation method
In vivo activation of macrophages
Principle
The basic principle of macrophage activation in vivo is that LPS, a complex of lipids and polysaccharides, is a major component of the cell wall of Gram-negative bacteria and is an endotoxin and an important group-specific antigen.LPS leads to the development of sepsis, septic shock, and even death by inducing activation of the monocyte-phagocyte system, which results in the release of large amounts of various pro-inflammatory cytokines, such as TNFα, IL-6, etc.TLR4 is the major pattern recognition receptor for the recognition of LPS by the monocyte-phagocyte system. TLR4 is the main pattern recognition receptor of monocyte-phagocyte system to recognize LPS.
Materials and Instruments
Animals: Move The basic process of in vivo activation of macrophages can be divided into the following steps: For more product details, please visit Aladdin Scientific website.
6-8 weeks old Kunming mice
Reagents:
Saline, 2 μg LPS (dissolved in 1 ml N
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Instrument:
Syringe.
(1) The mice were randomly divided into 2 groups, and were injected intraperitoneally with 1 ml of saline and 1 ml of LPS solution respectively. 12 hours later, the mice were decapitated and executed, and the peritoneal macrophages and peritoneal fluid were recovered respectively.
(2) The NO level of peritoneal fluid was detected by Griess method; TNF-α was determined by ELISA method.
(3) The peritoneal macrophages were cultured by wall-attachment method, and fluorescent microspheres were added to the culture medium at the same time, and cultured at 37 ℃ for 1-2 hours, and the phagocytosis ability of macrophages was observed by flow cytometry.
(4) The activation types of IL-12, IL-10, iNOS and Arg-1 can be analyzed at the same time, and each experiment can be repeated at least 3 times.