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Updated: Oct 22, 2025
Identifying Microglia and Peripheral Infiltrating Macrophages in the Injured Spinal Cords Using Flow Cytometry
Published on: June 24, 2025
S100-β aggravates spinal cord injury via activation of M1 macrophage phenotype
Keke Duan1, Shizhang Liu1, Zhi Yi1
1Shaanxi Provincial People's Hospital, the Third Affiliated Hospital of Xi'an Jiaotong University, Beilin District, Xi'an, Shaanxi, China.
Objectives:
S100-β has been identified as a sensitive biomarker in central nervous system injuries. However, the functions and mechanisms of S100-β are unknown in spinal cord injury.
Methods:
Spinal cord injury (SCI) mouse model was generated by surgical operation, microglia activation model was established by inducing BV-2 cells with LPS. The SCI model was evaluated by Basso-Beattie-Bresnahan (BBB) behavioral score, HE staining, and Nissl staining. The expression level of S100-β was detected by qRT-PCR, western blot, and immunofluorescence. qRT-PCR and western blot were used to detect the expression of iNOS and CD16. Pro-inflammatory cytokines TNF-α and IL-1β levels were detected by qRT-PCR and ELISA.
Results:
The expression of IL-1β, TNF-α, iNOS, and CD16 increased at 3rd day after SCI. In BV2 microglia, LPS treatment promoted the expression of S100-β, IL-1β, TNF-α, iNOS, and CD16. Knockdown of S100-β reduced the expression of iNOS stimulated by LPS. Over-expression of S100-β increased IL-1β and TNF-α, and S100-β inhibition suppressed IL-1β and TNF-α. In SCI mice, knockdown of S100-β attenuated the spinal cord injury and inhibited the expression of iNOS, IL-1β, and TNF-α.
Conclusions:
Down-regulation of S100-β could inhibit the pathogenesis of SCI and inhibit the activation of M1 macrophages. S100-β may be a useful diagnostic biomarker or therapeutic target for SCI.
Insights
Down-regulating S100-β protein may inhibit spinal cord injury (SCI) progression and M1 macrophage activation. S100-β shows potential as a diagnostic biomarker or therapeutic target for SCI.
Area of Science:
- Neuroscience
- Immunology
Background:
- S100-β is a sensitive biomarker for central nervous system injuries.
- The specific roles and mechanisms of S100-β in spinal cord injury (SCI) remain unclear.
Purpose of the Study:
- To investigate the function and mechanism of S100-β in spinal cord injury.
- To explore S100-β as a potential biomarker or therapeutic target for SCI.
Main Methods:
- Established a mouse model of spinal cord injury (SCI) and a microglia activation model.
- Assessed SCI severity using Basso-Beattie-Bresnahan (BBB) scores and histological staining.
- Quantified S100-β, iNOS, CD16, TNF-α, and IL-1β expression via qRT-PCR, western blot, and ELISA.
Main Results:
- S100-β expression and pro-inflammatory markers (IL-1β, TNF-α, iNOS, CD16) increased post-SCI and upon LPS stimulation in microglia.
- S100-β knockdown reduced LPS-induced iNOS expression and attenuated SCI, decreasing iNOS, IL-1β, and TNF-α levels.
- S100-β overexpression increased IL-1β and TNF-α, while S100-β inhibition suppressed them.
Conclusions:
- Down-regulation of S100-β inhibits SCI pathogenesis and M1 macrophage activation.
- S100-β presents potential as a diagnostic biomarker and therapeutic target for spinal cord injury.
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