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Disrupting RhoA activity by blocking Arhgef3 expression mitigates microglia-induced neuroinflammation post spinal
Liang Liao1, Zhan-Yang Qian2, Xin-Yu Li3
1Department of Orthopedics, Bengbu Third People's Hospital, 233000 Bengbu, China.
Abstract:
Excess inflammatory microglia activation deteriorates the pathological degree of spinal cord injury (SCI). We here employed microglia samples in vitro and murine model in vivo to trace the role of inhibition of Arhgef3 in inflammatory response post SCI. From the specimen analysis of lipopolysaccharide (LPS)-induced inflammatory microglia, we found that Arhgef3 expression was positively relative to microglia activation. In vitro, LPS caused the microglia inflammatory activation and induced upregulation of the Arhgef3 expression. Interestingly, presence of Arhgef3 could activate RhoA through promoting Rho GTPases, but silencing of Arhgef3 decreased RhoA activation and inhibited the microglia inflammation. Moreover, disruption of Arhgef3 inhibited the GTP-RhoA, resulted in a suppression of proinflammatory cytokines, and alleviated the LPS-elicited inflammatory genes expression. Moreover, artificially decreasing Arhgef3 expression remarkedly reduced ROS generation after LPS treatment. In vivo of a mouse mechanical contusion-induced SCI model, inhibition of Arhgef3 reduced the ratio of GTP-RhoA/Total-RhoA, and prevented SCI via mitigating the microglial inflammatory phenotype and decreased secondary neurological injury. Besides, inhibition of Arhgef3 prevented alleviated the degree of demyelination but did not affect neuronal regeneration. Meaningfully, absence of Arhgef3 improved mouse locomotor recovery post SCI. Taken together, Arhgef3 involves the microglial activation and inflammatory response following neural injury, and targeted disrupting of which may indicate a promising therapeutic direction in preventing SCI.
Insights
Inhibiting Arhgef3 reduces inflammatory microglia activation and secondary injury following spinal cord injury (SCI). This approach improves locomotor recovery in mice, suggesting Arhgef3 as a therapeutic target for SCI.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia activation exacerbates spinal cord injury (SCI) pathology.
- Arhgef3 expression correlates with microglia activation and inflammatory responses.
Purpose of the Study:
- To investigate the role of Arhgef3 in microglia activation and inflammation post-SCI.
- To evaluate the therapeutic potential of inhibiting Arhgef3 in a murine SCI model.
Main Methods:
- In vitro studies using lipopolysaccharide (LPS)-stimulated microglia.
- In vivo experiments using a mechanical contusion-induced SCI mouse model.
- Analysis of Arhgef3 expression, RhoA activation, inflammatory cytokines, ROS generation, and locomotor recovery.
Main Results:
- LPS upregulated Arhgef3, which activated RhoA and promoted microglia inflammation.
- Arhgef3 inhibition decreased RhoA activation, suppressed pro-inflammatory cytokines and ROS generation, and alleviated inflammatory gene expression.
- In vivo, Arhgef3 inhibition mitigated microglial inflammation, reduced secondary neurological injury, and improved locomotor recovery, without affecting neuronal regeneration.
Conclusions:
- Arhgef3 plays a critical role in microglia activation and inflammatory response after neural injury.
- Targeting Arhgef3 represents a promising therapeutic strategy for preventing and treating spinal cord injury.
