Circ0005512 promotes microglia/macrophage pyroptosis after spinal cord injury in female rats / 中国组织工程研究

ABSTRACT

BACKGROUND:Neuroinflammation is an important factor leading to secondary spinal cord injury,and microglia/macrophage pyroptosis is a significant part of post-spinal cord injury neuroinflammation.Studies have shown that microglia/macrophage undergoes pyroptosis after spinal cord injury,but the regulatory mechanism of circular RNA(circRNA)in microglia/macrophage pyroptosis after spinal cord injury remains unclear. OBJECTIVE:To investigate the role and mechanism of circRNA0005512 in regulating microglia/macrophage pyroptosis after spinal cord injury. METHODS:Female Wistar rats were divided into sham group and spinal cord injury group.Motor function was evaluated using the Basso,Beattie,and Bresnahan(BBB)scale.Cavity volume was assessed by hematoxylin-eosin staining.Differential expression of circRNA in spinal cord tissue was screened using RNA-sequencing and circ0005512 was validated by real-time PCR.Immunofluorescence,western blot assay,ELISA,and real-time PCR were performed to detect cell pyroptosis in the rats and lipopolysaccharide-induced microglial cell line HAPI cell models.Gene knockdown was used to confirm the regulatory role of circRNA0005512 in microglia/macrophage pyroptosis. RESULTS AND CONCLUSION:(1)Seven days after spinal cord injury,evident cavities were observed at the injury site.Immediately after spinal cord injury,the motor function of rats was completely lost.Over time,the motor function of rats in the spinal cord injury group gradually partially recovered,and there was a significant difference in BBB scores compared to the sham group.(2)Circ0005512 was significantly upregulated according to the results of the RNA-sequencing and confirmed in both the animal and cell models.(3)Immunofluorescence,western blot assay,real-time PCR,and ELISA confirmed the significant upregulation of cell pyroptosis markers(NLRP3,GSDMD,and caspase-1)in spinal cord injury tissue and lipopolysaccharide-induced HAPI cells.(4)In the cell model,knockdown of circ0005512 resulted in significantly decreased levels of cell pyroptosis marker-NLRP3.(5)The results above indicate that circ0005512 promotes pyroptosis in microglia/macrophages after spinal cord injury.