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Updated: Jan 29, 2026
Three-dimensional Confocal Analysis of Microglia/macrophage Markers of Polarization in Experimental Brain Injury
Published on: September 4, 2013
Effects of hydrogen on polarization of macrophages and microglia in a stroke model
Ke Ning1, Wen-Wu Liu2, Jun-Long Huang1
1Department of Navy Aeromedicine, Faculty of Naval Medicine, Navy Medical University, Shanghai, China.
Abstract:
It has been confirmed that inflammation plays an important role in the pathogenesis of ischemic stroke. The polarization of microglia as an important participant in the inflammation following stroke is also found to be involved in stroke. This study aimed to investigate the effects of hydrogen gas on the polarization of macrophages/microglia in vitro. Raw264.7 cells were treated with lipopolysaccharides and then exposed to hydrogen. The microglia were treated with the supernatant from oxygen and glucose deprivation-treated neurons and then exposed to hydrogen. The phenotypes of Raw 264.7 cells and microglia were determined by flow cytometry, and cell morphology was observed. Results showed lipopolysaccharides significantly increased the M1 macrophages, and the supernatant from oxygen and glucose deprivation-treated neurons dramatically elevated the proportion of M1 microglia, but both treatments had little influence on the M2 cells. In addition, hydrogen treatment significantly inhibited the increase in M1 cells, but had no influence on M2 ones. Our findings suggest that the neuroprotection of hydrogen may be related to its regulation of microglia in the nervous system after stroke.
Insights
Hydrogen gas may protect against ischemic stroke by regulating microglia polarization. This study found hydrogen inhibited the M1 (pro-inflammatory) macrophage/microglia phenotype in vitro, suggesting a potential therapeutic mechanism.
Area of Science:
- Neuroscience
- Immunology
- Biochemistry
Background:
- Inflammation is crucial in ischemic stroke pathogenesis.
- Microglia polarization significantly impacts stroke-related inflammation.
- Understanding microglia modulation is key for stroke therapeutics.
Purpose of the Study:
- To investigate the effect of hydrogen gas on macrophage and microglia polarization in vitro.
- To determine if hydrogen influences the M1/M2 phenotypes relevant to stroke inflammation.
Main Methods:
- Utilized Raw264.7 cells and primary microglia cultures.
- Induced M1 polarization using lipopolysaccharides and neuronal supernatant.
- Exposed cells to hydrogen gas and analyzed phenotypes via flow cytometry.
Main Results:
- Lipopolysaccharide and neuronal supernatant increased M1 macrophages/microglia.
- Hydrogen gas treatment significantly reduced M1 cell proportions.
- Hydrogen did not significantly alter M2 (anti-inflammatory) cell populations.
Conclusions:
- Hydrogen gas may exert neuroprotection in ischemic stroke by modulating microglia polarization.
- Hydrogen's anti-inflammatory effect is linked to the inhibition of M1 phenotype.
- Further research into hydrogen therapy for stroke is warranted.
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