| Objective: To study the effects and mechanism of probenecid(PROB)in the acti vation of NACHT-LRR-PYD-containing protein 1(NLRP1)inflammasome in acute s pinal cord injury rat.Methods:The spinal cord injury was produced by the clip compression model de scribed by Rivlin and Tator.Ninety male adult Sprague-Dawley(SD)rats,220-250 g,were randomly divided into sham-operated group(Sham group),spinal cord inj ury group(SCI group)and PROB group(PROB group).The vertebral plates of S ham group were resected.The PROB was intraperitoneal injection immediately af ter spinal cord injury,5 mg/kg every 12 h for 3 days,and the normal saline was used as solvent control.(1)The locomotor function of hind limbs was evalucate d by Basso-Beattle-Bresnahan(BBB)score on the 1,3,7,14,21 day after spinal cord injury.(2)The expression of Caspase-1,XIAP were detected by western-bl ot,and the samples were obtained on 6 h,24 h,72 h after spinal cord injury.(3)The expression levels of Caspase-1,IL-1? were detected by immunochemistry sta in,and the samples were obtained on 6 h after spinal cord injury;(4)The histop athological changes of spinal cords were observed by HE staining,and the sample s were obtained on 72 h after spinal cord injury..Results:(1)The BBB scores of Sham group at different time points were higher than SCI group and PROB group,while the scores of PROB group on 3 day,7day,14 day and 21 day after injure were higher than SCI group with statistical significance(p<0.05).(2)Western blot results showed that the expression of Cas pase-1 in SCI group was significantly higher than Sham group in the time-depend ent manners,which was highest in 72 h after injury,while the expression of Cas pase-1 in PROB group was lower than SCI group.The expression of XIAP in Sh am group was observably higher than SCI group and PROB group,and the expre ssion of XIAP in SCI group was reduced in a time-dependent manner,which was lowest in 72 h after injury.The expression of XIAP in PROB group was higher than SCI group with statistical significance(p<0.05).(3)Immunohistochemistry staining revealed that the expression of Caspase-1 and IL-1? of SCI group were sig nificantly higher than Sham group,whereas the expression of Caspase-1 and IL-1? in PROB group were significantly lower than SCI group with statistical signific ance(p<0.05).(4)The HE staining results showed that compared with SCI grou p,PROB group showed less neutrophil infiltration,less edema,fewer neuronal de generation and necrosis and smaller spinal cavity area.Conclusion: PROB has positive effects in acute spinal cord injury rat,which co uld reduce the secondary spinal cord injury by inhibiting the activation of NLRP1 inflammasome,and it is related to the cleavage of XIAP and the activation of Ca spase-1. |
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