大鼠原代神经元中依达拉奉对糖氧剥夺损伤的保护及对ROS/TXNIP/NLRP3通路的影响

Effects of edaravone on protecting glucose-oxygen deprivation injury and the reactive oxygen species/thioredoxin interacting protein/NOD-like receptor associated protein 3 pathway in rat primary neurons

目的探讨依达拉奉后处理对神经元糖氧剥夺损伤的保护作用及对活性氧(ROS)/硫氧还蛋内结合蛋白(TXNIP)/NOD样受体相关蛋白3(NOD-like receptor associated protein 3,NLRP3)信号通路的影响。方法培养新生SD大鼠皮质神经元,经免疫荧光染色鉴定后随机分为神经元组、依达拉奉组、糖氧剥夺组、糖氧剥夺+依达拉奉组4组。各组神经元在完成处理后以CCK-8试剂盒检测神经元活力;以细胞凋亡试剂盒检测神经元凋亡情况;以ELISA试剂盒检测神经元ROS产生情况;以蛋白免疫印迹法检测ROS/TXNIP/NLRP3信号通路相关蛋白的表达水平。结果神经元缺糖缺氧后细胞活力显著降低,依达拉奉处理后又显著上升,差异有统计学意义(P<0.01);糖氧剥夺后,神经元凋亡水平、ROS含量、ROS/TXNIP/NLRP3通路相关IL-1β、TXNIP、IL-18、Caspase-1、ASC和NLRP3蛋白表达水平升高,依达拉奉处理后又显著下降,差异有统计学意义(P<0.01)。结论依达拉奉对神经元糖氧剥夺损伤具有保护作用,可能与其对R0S/TXN1P/NLKP3信号通路的抑制有关。

Objective To explore the protective effects of edaravone postconditioning on cerebral hypoxiainjury in rats and its effect on the reactive oxygen species(ROS)/thioredoxin interacting protein(TXNIP)/NOD-like receptor-associated protein3(NLRP3)signaling pathway.Methods Neonatal SD rat cortical neurons were cultured and identified by immunofluorescence staining and randomly divided into 4 groups:neuron group,edaravone group,glucose-oxygen deprivation group,glucose-oxygen deprivation+edaravone group.Neurons in each group were tested for neuronal viability by CCK-8 kit after completion of treatment;neuronal apoptosis by apoptosis kit;neuronal ROS production by ELISA kit;and the expression levels of neuronal proteins by Western blotting.Results The cell viability was significantly decreased after glucose deprivation and hypoxia,and increased significantly after edaravone treatment,and the difference was statistically significant(P<0.01);after glucose deprivation,the neuronal apoptosis level,ROS content,IL-1βprotein expression level,TXNIP protein expression level,IL-18 protein expression level,caspase-1 protein expression level,ASC protein expression level and NLRP3 protein expression level were significantly increased,and decreased significantly after edaravone treatment,and the difference was statistically significant(P<0.01).Conclusion Edaravone has a neuronal protective effect,which may be related to the inhibition of the ROS/TXNIP/NLRP3 signaling pathway.