Related Experiment Video
Updated: Nov 5, 2025
Metabolic Characterization of Polarized M1 and M2 Bone Marrow-derived Macrophages Using Real-time Extracellular Flux Analysis
Published on: November 28, 2015
sDR5-Fc inhibits macrophage M1 polarization by blocking the glycolysis
Guang-Yao Zhai1, Shu-Yan Qie2, Qian-Yun Guo1
1Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart Lung and Blood Vessel Disease, Beijing Key Laboratory of Precision Medicine of Coronary Atherosclerotic Disease, Clinical Center for Coronary Heart Disease, Capital Medical University, Beijing, China.
Background:
M1 polarization of macrophages is an important pathological process in myocardial ischemia reperfusion injury, which is the major obstacle for the treatment of acute myocardial infarction. Currently, the strategies and mechanisms of inhibiting M1 polarization are poorly explored. This study aims to investigate the role of soluble death receptor 5-Fc (sDR5-Fc) in regulating M1 polarization of macrophages under extreme conditions and explore the mechanisms from the aspect of glycolysis.
Methods:
Extreme conditions were induced in RAW264.7 cells. Real-time quantitative polymerase chain reaction and western blot were used to detect the expression of mRNA and proteins, respectively. Cell counting kit-8 was used to investigate the proliferation activity of cells. Expression levels of inflammatory cytokines were determined by enzyme-linked immunosorbent assay.
Results:
We found that sDR5-Fc rescues the proliferation of macrophages under extreme conditions, including nutrition deficiency, excessive peroxide, and ultraviolet irradiation. In addition, administration of sDR5-Fc inhibits the M1 polarization of macrophages induced by lipopolysaccharide (LPS) and interferon-gamma (IFN-γ), as the expression of M1 polarization markers CD86, CXC motif chemokine ligand 10, matrix metalloproteinase 9, and tumor necrosis factor-α, as well as the secretion of inflammatory factors interleukin (IL)-1β and IL-6, were significantly decreased. By further investigation of the mechanisms, the results showed that sDR5-Fc can recover the LPS and IFN-γ induced pH reduction, lactic acid elevation, and increased expression of hexokinase 2 and glucose transporter 1, which were markers of glycolysis in macrophages.
Conclusions:
sDR5-Fc inhibits the M1 polarization of macrophages by blocking the glycolysis, which provides a new direction for the development of strategies in the treatment of myocardial ischemia reperfusion injury.
Insights
Soluble death receptor 5-Fc (sDR5-Fc) inhibits M1 macrophage polarization by blocking glycolysis. This finding offers a novel therapeutic strategy for myocardial ischemia reperfusion injury.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- M1 macrophage polarization is key in myocardial ischemia reperfusion injury, hindering acute myocardial infarction treatment.
- Current strategies to inhibit M1 polarization and its mechanisms remain underexplored.
Purpose of the Study:
- Investigate soluble death receptor 5-Fc (sDR5-Fc) role in regulating M1 macrophage polarization under stress.
- Explore the underlying mechanisms focusing on glycolysis.
Main Methods:
- Induced extreme conditions in RAW264.7 cells.
- Utilized real-time quantitative polymerase chain reaction, western blot, cell counting kit-8, and enzyme-linked immunosorbent assay.
- Assessed proliferation, M1 polarization markers, inflammatory cytokine secretion, and glycolysis indicators.
Main Results:
- sDR5-Fc rescued macrophage proliferation under various extreme conditions.
- sDR5-Fc inhibited M1 polarization markers (CD86, CXCL10, MMP-9, TNF-α) and inflammatory cytokines (IL-1β, IL-6).
- sDR5-Fc reversed LPS/IFN-γ-induced reduction in pH, increased lactic acid, and elevated glycolysis markers (HK2, GLUT1).
Conclusions:
- sDR5-Fc inhibits M1 macrophage polarization by suppressing glycolysis.
- This presents a new therapeutic avenue for myocardial ischemia reperfusion injury.
