DDX3X/NF-κB通路介导蛛网膜下腔出血小鼠早期神经元凋亡
DDX3X/NF-κB pathway mediates early neurons apoptosis in mice with subarachnoid hemorrhage
郝广志 1郇宇 1韩雨薇 1董玉书 1梁国标1
作者信息
- 1. 中国人民解放军北部战区总医院神经外科,沈阳 110016
- 折叠
摘要
目的:研究DDX3X/NF-κB通路在蛛网膜下腔出血(SAH)小鼠早期神经元凋亡过程中的作用.方法:利用颈内动脉穿刺的方法构建SAH小鼠模型,对小鼠进行神经功能评分.使用表达DDX3X靶向shRNA的重组慢病毒(Lv-shDDX3X)预先敲低脑内DDX3X的表达,或者通过NF-κB抑制剂NF-κB-IN-1(简称IN-1)抑制NF-κB信号通路,利用Western Blot检测小鼠皮质DDX3X和NF-κB(p65)的表达,通过TUNEL/NeuN染色检测各组小鼠大脑皮质神经元的凋亡.结果:SAH术后24d小鼠神经功能显著障碍(P＜0.05),皮质中DDX3X表达显著增加而NF-KB(p65)的表达显著降低(P＜0.05).预先敲低DDX3X后,小鼠神经功能显著恢复,NF-KB(p65)蛋白表达显著高于SAH组(P＜0.05);当在敲低DDX3X表达的同时使用IN-1抑制NF-κB活性,则小鼠神经功能恢复不明显.TUNEL/NeuN染色显示敲低DDX3X表达后小鼠脑组织中TUNEL阳性的死亡神经元数量少于SAH组(P＜0.05),而如果在敲低DDX3X表达的同时使用IN-1抑制NF-κB活性,则TUNEL阳性的神经元数量减少不明显.结论:DDX3X/NF-κB通路介导了 SAH后早期脑损伤小鼠的细胞死亡.
Abstract
Objective:To study the role of DDX3X/NF-κB pathway in early neuronal apoptosis in subarachnoid hemorrhage(SAH)mice.Methods:The mouse model of SAH was established by internal carotid artery puncture,and the neurological function score of the mice was evaluated.The DDX3X expression was knocked down using recombinant lentivirus expressing DDX3X targeted shRNA(Lv-shDDX3X),or the NF-κB pathway was inhibited by NF-κB-IN-1(IN-1).Western Blot was used to detect the expression of DDX3X and NF-κB(p65)in mouse cortex.TUNEL/NeuN staining was used to detect the apoptosis of cerebral cortex neurons.Results:Twenty-four hours after SAH operation,the neurological function of mice was significantly impaired(P＜0.05).While the expression of DDX3X was signifi-cantly increased and the expression of NF-κB(p65)was significantly decreased in the cortex(P＜0.05).When the DDX3X expression is knocked down firstly,then SAH surgery is performed.The neurological function of mice was sig-nificantly recovered,and the expression of NF-κB(p65)protein was significantly higher than that in SAH group(P＜0.05);If the NF-κB activity was inhibited by IN-1 while DDX3X knockdown,there is no significant recovery of neuro-logical function in SAH mice.TUNEL/NeuN staining showed that the number of TUNEL-positive neurons in the brain tissue after DDX3X knockdown was less than that in the SAH group(P＜0.05),while the number of TUNEL-positive neurons was not significantly reduced when IN-1 was used to inhibit NF-κB activity at the same time of DDX3X knock-down.Conclusion:DDX3X/NF-κB mediated cell death in mice with early brain injury after SAH.
关键词
DEAD-box/RNA解旋酶3(DDX3X)/NF-κB/蛛网膜下腔出血/早期脑损伤/凋亡/小鼠Key words
DEAD-box helicase 3 X-linked(DDX3X)/NF-κB/subarachnoid hemorrhage(SAH)/early brain in-jury(EBI)/apoptosis/mouse引用本文复制引用
出版年
2024
NETL
NSTL
万方数据