右美托咪定对小鼠海马神经元缺氧复氧时Src介导NR2A酪氨酸磷酸化水平的影响  

Effect of dexmedetomidine on Src-mediated NR2A tyrosine phosphorylation in hippocampal neurons subjected to hypoxia-reoxygenation in mice

作　　者：于向洋[1] 贾莉莉[2] 喻文立[2] 杜洪印[2] Yu Xiangyang;Jia Lili;Yu Wenli;Du Hongyin(Tianjin Medical University First Central Clinical College、Tianjin 300192, China;Department of Anesthesiology, Tianjin First Central Hospital, Tianjin 300192, China)

机构地区：[1]天津医科大学一中心临床学院,300192 [2]天津市第一中心医院麻醉科,300192

出　　处：《中华麻醉学杂志》2018年第10期1249-1252,共4页Chinese Journal of Anesthesiology

基　　金：天津市卫生行业重点攻关项目(16KG101);天津市卫生和计划生育委员会中医中西医结合科研课题(2017056).

摘　　要：目的评价右美托咪定对小鼠海马神经元缺氧复氧时Src介导NR2A酪氨酸磷酸化水平的影响。方法拉颈处死孕18d的C57小鼠,剖腹取胎鼠,分离海马神经元。将神经元接种于培养液中,培养至第7天,采用随机数字表法分为3组(n=30):对照组(C组)、缺氧复氧组(H/R组)和右美托咪定组(D组)。H/R组和D组缺氧4h复氧24h制备小鼠海马神经元缺氧复氧损伤模型;D组给予终浓度为1μmol/L的右美托咪定孵育4h,随后制备模型。于复氧24h时采用MTT法检测海马神经元活力,TUNEL染色法观察海马神经元凋亡情况,计算细胞凋亡指数,Westernblot法检测海马神经元c-Src、p-SrcY416、NR2A、p-NR2AY1325和cleavedcaspase-3表达水平。结果与C组比较,H/R组海马神经元活力降低,凋亡指数升高,p-SrcY416、p-NR2AY1325和cleavedcaspase-3表达上调(P<0.05);与H/R组比较,D组海马神经元活力升高,凋亡指数降低,p-SrcY416、p-NR2AY1325和cleavedcaspase-3表达下调(P<0.05)。3组c-Src和NR2A表达比较差异无统计学意义(P>0.05)。结论右美托咪定可抑制缺氧复氧小鼠海马神经元凋亡,其机制可能与降低Src介导NR2A酪氨酸磷酸化水平有关。Objective To evaluate the effect of dexmedetomidine on Src-mediated NR2A tyrosine phosphorylation in hippocampal neurons subjected to hypoxia-reoxygenation(H/R)in mice. Methods C57 mice at 18 days of gestation were sacrificed by pulling neck, fetal mice were obtained by caesarean section, and hippocampal neurons were isolated.Neurons were cultured in culture medium for 7 days and then divided into 3 groups(n=30 each)using a random number table method: control group(group C), H/R group and dexmedetomidine group(D group). Hippocampal neurons were exposed to 95% N2-5% CO2 in an incubator at 37 ℃ for 4 h followed by reoxygenation with 95% O2-5% CO2 for 24 h to establish the model of H/R.Dexmedetomidine was given at a final concentration of 1 μmol/L, and neurons were incubated for 4 h before establishing the model.The viability of hippocampal neurons was measured by MTT assay, TUNEL staining was used to observe apoptosis in hippocampal neurons, and the expression of c-Src, phosphorylated Src Y416(p-Src Y416), NR2A, phosphorylated NR2A Y1325(p-NR2A Y1325)and cleaved caspase-3 was determined by Western blot.Apoptosis index was calculated. Results Compared with group C, the viability of hippocampal neurons was significantly decreased, apoptosis index was increased, and the expression of p-Src Y416, p-NR2A Y1325 and cleaved caspase-3 was up-regulated in group H/R(P<0.05). Compared with group H/R, the viability of hippocampal neurons was significantly increased, apoptosis index was decreased, and the expression of p-Src Y416, p-NR2A Y1325 and cleaved caspase-3 was down-regulated in group D(P<0.05). There was no significant difference in the expression of c-Src and NR2A among three groups(P>0.05). Conclusion Dexmedetomidine can inhibit apoptosis in hippocampal neurons subjected to H/R, and the mechanism may be associated with decreasing Src-mediated NR2A tyrosine phosphorylation in mice.

关 键 词：右美托咪啶 缺氧 海马 神经元 src族激酶类 受体 N-甲基-D-天冬氨酸 酪氨酸 

分 类 号：R614[医药卫生—麻醉学]