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Updated: Jan 12, 2026
Author Spotlight: Microglia Research on Spinal Cord Heterogeneity and Purification
Published on: September 22, 2023
Microglial SLC25A28 Knockout Mitigates Spinal Cord Injury in Mice by Inhibiting Heme Synthesis and Subsequent NOX2
Huangtao Chen1, Shaochun Guo2, Yanglan Mi3,4
1Department of Neurosurgery, The Second Affiliated Hospital of Xi'an Jiao Tong University, Xi'an, Shaanxi, China.
Background:
Microglial overactivation-driven neuroinflammation exacerbates secondary damage after spinal cord injury (SCI), but the role of mitochondrial iron metabolism in this process is not well understood. This study investigates the function of the mitochondrial iron transporter solute carrier family 25 member 28 (SLC25A28) in post-SCI neuroinflammation.
Methods:
Microglia-specific SLC25A28 knockout (A28-MGKO) mice were generated by crossing SLC25A28flox/flox mice with Cx3cr1-CreERT2 mice and subjected to clip-compression spinal cord injury (SCI) at the T9 level. Motor recovery was evaluated using the Basso Mouse Scale (BMS), while histological and biochemical assessments including hematoxylin-eosin and Nissl staining, Iba1 immunohistochemistry, Evans blue permeability, and tissue water content were performed to evaluate lesion severity, neuronal survival, microglial activation, and blood-spinal cord barrier integrity. In vitro, primary microglia isolated from A28-MGKO mice and BV2 cells with SLC25A28 overexpression were used to investigate mitochondrial iron homeostasis, heme biosynthesis, and NOX2-mediated oxidative stress. Mitochondrial iron content was quantified using a ferrozine-based assay and Mito-FerroGreen staining, while ROS production, cytokine release, and inflammatory signaling were analyzed by fluorescence imaging, ELISA, and Western blotting under pharmacological modulation of heme synthesis and NOX2 activity.
Results:
We found that SLC25A28 deficiency reduced spinal cord edema, blood-spinal cord barrier disruption, and motor deficits. Mechanistically, SLC25A28 knockout suppressed mitochondrial iron accumulation, inhibited heme synthesis, and reduced NOX2-mediated oxidative stress. However, SLC25A28 overexpression enhanced mitochondrial iron overload and NOX2-driven inflammation, which could be reversed by pharmacological blockade of NOX2 or heme synthesis. Restoration of heme synthesis in A28-MGKO microglia attenuated the anti-inflammatory effects of SLC25A28 knockout.
Conclusion:
These findings demonstrate that microglial SLC25A28 regulates neuroinflammation and functional recovery after SCI by promoting mitochondrial iron-dependent heme synthesis and NOX2 activation. Targeting the SLC25A28-heme-NOX2 axis may provide a novel therapeutic approach for SCI.
Insights
Microglial solute carrier family 25 member 28 (SLC25A28) regulates spinal cord injury (SCI) neuroinflammation. Targeting the SLC25A28-heme-NOX2 pathway may offer new therapeutic strategies for SCI recovery.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglial overactivation drives neuroinflammation, worsening spinal cord injury (SCI) secondary damage.
- The role of mitochondrial iron metabolism in SCI-related neuroinflammation remains unclear.
- This study focuses on the mitochondrial iron transporter solute carrier family 25 member 28 (SLC25A28) in post-SCI neuroinflammation.
Purpose of the Study:
- To investigate the function of SLC25A28 in microglial response to SCI.
- To elucidate the mechanisms linking mitochondrial iron metabolism to neuroinflammation after SCI.
- To explore the therapeutic potential of targeting the SLC25A28 pathway in SCI.
Main Methods:
- Generated microglia-specific SLC25A28 knockout (A28-MGKO) mice for SCI studies.
- Assessed motor recovery using the Basso Mouse Scale (BMS) and evaluated lesion severity via histological and biochemical analyses.
- Utilized primary microglia and BV2 cells to examine mitochondrial iron, heme synthesis, and NOX2-mediated oxidative stress in vitro.
Main Results:
- SLC25A28 deficiency reduced SCI-induced edema, blood-spinal cord barrier disruption, and motor deficits.
- Knockout of SLC25A28 suppressed mitochondrial iron accumulation, inhibited heme synthesis, and decreased NOX2-mediated oxidative stress.
- Overexpression of SLC25A28 exacerbated inflammation, which was reversible by blocking NOX2 or heme synthesis.
Conclusions:
- Microglial SLC25A28 promotes SCI neuroinflammation and impairs functional recovery via mitochondrial iron-dependent heme synthesis and NOX2 activation.
- The SLC25A28-heme-NOX2 axis represents a potential therapeutic target for SCI.
- Modulating microglial mitochondrial iron metabolism offers a novel strategy for SCI treatment.
