异丙酚联合缺氧诱导未成熟大鼠发生认知功能障碍的机制  

Mechanism of propofol combined with hypoxia inducing cognitive dysfunction in immature rats

作　　者：刘亚男[1] 孙颜 李东兴[1] LIU Ya-nan;SUN Yan;LI Dong-xing(Department of Anesthesiology,Handan Central Hospital of Hebei Province,Handan Hebei 056000,China)

机构地区：[1]邯郸市中心医院麻醉科,河北邯郸056000

出　　处：《临床和实验医学杂志》2021年第20期2135-2140,共6页Journal of Clinical and Experimental Medicine

基　　金：2019年河北省医学科学研究课题计划(编号:20191849)。

摘　　要：目的探讨异丙酚联合缺氧诱导未成熟大鼠发生认知功能障碍的机制。方法24只Wistar大鼠幼崽,日龄7 d,体重(14±1)g,置于一个塑料腔中,随机数字表法分成3组:对照组(21%氧浓度暴露1.5 h,n=8),缺氧诱导组[(5±0.5)%氧浓度暴露1.5 h,n=8]和联合诱导组[3%的异丙酚+(5±0.5)%氧浓度暴露1.5 h,n=8]。总气体流量为2 L/min。使用气体分析系统测量氧气和麻醉剂的含量。通过莫里斯水迷宫测试以评估空间学习和记忆能力。通过实时荧光定量聚合酶链式反应测量海马中促炎细胞因子[肿瘤坏死因子α(TNF-α)和白细胞介素-1β(IL-1β)]的水平。通过免疫荧光染色检测大鼠海马中Bax、caspase-3和Bcl-2表达水平。通过神经元特异性核蛋白(NeuN)染色检测神经元阳性细胞数量。通过蛋白质免疫印迹检测在Ser1412处神经元一氧化氮合酶(nNOS)的磷酸化(pnNOS-Ser1412)和pnNOS-Ser847和过氧化物酶体增殖物激活受体(PPAR)-γ蛋白表达水平。结果随着时间的增加,3组逃避潜伏期均降低,游泳速度均增加,差异有统计学意义(P<0.05)。与对照组相比,缺氧诱导组和联合诱导组逃避潜伏期增加(P<0.05),而游泳速度差异无统计学意义(P>0.05)。与对照组相比,缺氧诱导组和联合诱导组的TNF-α和IL-1β的mRNA、Bax和caspase-3的表达、pnNOS-Ser1412的蛋白水平均增加,Bcl-2表达水平、神经元阳性细胞数量、pnNOS-Ser847和PPAR-γ蛋白表达水平均降低,差异均有统计学意义(P<0.05)。与缺氧诱导组相比,联合诱导组逃避潜伏期、TNF-α和IL-1β的mRNA、Bax和caspase-3的表达、pnNOS-Ser1412的蛋白水平均增加,Bcl-2蛋表达水平、神经元阳性细胞数量、pnNOS-Ser847和PPAR-γ蛋白表达水平均降低,差异均有统计学意义(P<0.05)。结论异丙酚联合缺氧诱导未成熟大鼠,通过增加海马神经炎症以及pnNOS-Ser1412表达,降低pnNOS-Ser847和PPAR-γ表达,导致神经元损伤和凋亡,进一步加剧认知功能下降。Objective To explore the mechanism of propofol combined with hypoxia inducing cognitive dysfunction in immature rats.Methods Twenty four Wistar rat pups,7 days old,weight(14±1)g were placed in a plastic cavity,and the experiment was divided into 3 groups according to the random number table method:control group(21%oxygen concentration exposure for 1.5 h,n=8),hypoxia induction group[(5%±0.5)%oxygen concentration exposure for 1.5 h,n=8]and combined induction group[3%propofol+(5±0.5)%oxygen concentration exposure for 1.5 h,n=8].The total gas flow rate was 2 L/min.The content of oxygen and anesthetics was measured using a gas analysis system.Spatial learning and memory abilities were tested and evaluated through the Morris Water Maze.The levels of pro-inflammatory cytokines[tumor necrosis factor alpha(TNF-α),interleukin-1β(IL-1β)]in the hippocampus were measured by qRT-PCR.The expression levels of Bax,caspase-3 and Bcl-2 in rat hippocampus were detected by immunofluorescence staining.The number of neuron-positive cells was detected by neuron specific nuclear protein(NeuN)staining.Phosphorylation of neuronal nitric oxide synthase(nNOS)at Ser1412(pnNOS-Ser1412)and the protein expression levels of pnNOS-Ser847 and peroxisome proliferator activated receptor-γ(PPAR-γ)protein expression levels at Ser1412 were detected by Western blotting.Results With the increase of time,the escape latency of the three groups decreased,and the swimming speed increased,and the differences were statistically significant(P<0.05).Compared with the control group,the escape latency of hypoxia induction group and combined induction group were increased,the differences were statistically significant(P<0.05),and there was no difference in swimming speed(P>0.05).Compared with the hypoxia induction group,the escape latency,the expression of TNF-αand IL-1βmRNA,Bax and caspase-3,and the level of pnNOS-Ser1412 protein were significantly increased in the combined induction group,while the expression level of Bcl-2 protein,the number of neuron-p

关 键 词：未成熟大鼠 异丙酚 缺氧 海马 神经元损 认知功能 

分 类 号：R614[医药卫生—麻醉学]