异丙酚通过c-Jun氨基末端激酶(c-Jun N-terminal kinase)信号途径诱导神经元细胞凋亡  

Propofol-induced Apoptosis in Neuronal Cells by Means of JNK Signaling Pathway

作　　者：展希[1] 杨林珠[1] 雷常成[1] Zhan Xi;Yang Linzhu;Lei Changcheng(First Affiliated Hospital of Kunming Medical University,Kunming,650032)

机构地区：[1]昆明医科大学第一附属医院,昆明650032

出　　处：《基因组学与应用生物学》2020年第7期3400-3406,共7页Genomics and Applied Biology

摘　　要：为了阐明线粒体途径在异丙酚诱导神经毒性中的作用机制,本研究通过人类胚胎干细胞(human embryonic stem cells,hESC)诱导分化获得神经元细胞;使用TUNEL染色评估hESC衍生神经元细胞的细胞死亡情况,结合Annexin-V/7-AAD分析细胞凋亡率变化;并通过Western blotting对神经元细胞中凋亡相关蛋白因子及JNK途径蛋白因子表达进行评估。结果显示,异丙酚以剂量和时间依赖性方式诱导hESC衍生神经元细胞的细胞死亡;JNK在异丙酚处理的神经元细胞中被强烈激活,并且Western blotting和Annexin V-PE/7-AAD分析证实,引发JNK活化的异丙酚剂量与诱导神经元凋亡的剂量一致。此外,活性氧簇(reactive oxygen species,ROS)清除剂N-乙酰基-L-半胱氨酸(N-acetyl-L-cysteine,NAC)可缓解异丙酚诱导的磷酸化JNK(Phosphorylation JNK,p-JNK)线粒体和细胞核易位,表明ROS的积累在异丙酚诱导的JNK活化中发挥作用;同时,Annexin V-PE/7-AAD分析显示,NAC和SP600125(JNK抑制剂)处理可显著下调异丙酚诱导的神经元凋亡率,分别从(47.13±1.15)%降至(30.11±0.27)%和(27.122±2.25)%。由此表明,JNK信号传导通过线粒体易位和促凋亡基因的转录,在异丙酚诱导hESC衍生神经元细胞中发挥关键作用。This article aims to elucidate the mechanism of action of mitochondrial pathway in propofol-induced neurotoxicity.Neuronal cells were obtained by differentiation induced by hESC;TUNEL staining was used to evaluate the cell death of hESC-derived neurons;And apoptosis was measured by Annexin-V/7-AAD.The expression of apoptosis-related protein factors and JNK pathway protein factors in neuronal cells was evaluated by Western blotting.Propofol induced cell death in hESC-derived neuronal cells in a dose-and time-dependent manner;JNK was strongly activated in propofol-treated neuronal cells.Western blotting and Annexin V-PE/7-AAD analysis confirmed that the dose of propofol that triggers JNK activation is consistent with the dose that induces neuronal apoptosis.In addition,ROS accumulation plays a role in propofol-induced JNK activation,as propofol-induced phosphorylation of JNK(p-JNK)mitochondria and nuclear translocations can be alleviated by ROS scavenger NAC;meanwhile,Annexin V-PE/7-AAD analysis showed that treatment with NAC or SP600125(JNK inhibitor)significantly down-regulated propofol-induced neuronal apoptosis from(47.13±1.15)%to(30.11±0.27)%or(27.122±2.25)%,respectively.JNK signaling plays a key role in propofol-induced hESC-derived neuronal cells through transcription of mitochondrial translocation and proapoptotic genes.

关 键 词：异丙酚 JNK ROS 细胞凋亡 神经毒性 

分 类 号：R96[医药卫生—药理学]