H2对大鼠全脑缺血再灌注损伤后海马CA1区神经元及树突棘的保护作用  

Protective effect of hydrogen gas on neurons and dendritic spines of hippocampns CA1 region in rats after global cerebral ischemia/reperfusion injury

作　　者：谭永星[1] 袁楠楠[2] 夏裕宁 张鑫磊[2] 梁维[3] 魏佑震[4] 

机构地区：[1]桂林医学院附属医院重症医学科,桂林541001 [2]桂林医学院研究生院,桂林541000 [3]桂林医学院附属医院麻醉科,桂林541001 [4]同济大学东方转化医学研究中心,上海200120

出　　处：《中华神经医学杂志》2016年第11期1091-1097,共7页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（81260206）;广西壮族自治区自然科学基金（2013GXNSFAA019155、2015GXNSFAA139130）;广西壮族自治区卫生厅课题（Z2012402、Z2013497）

摘　　要：目的探讨吸入高浓度H2对大鼠全脑缺血再灌注损伤后海马CA1区神经元及树突棘的保护作用及可能机制。方法健康雄性SD大鼠120只按随机数字表法分为3组：假手术组、模型组和治疗组（n=40），假手术组吸入含67％N2和33％O2的混合气体，模型组、治疗组采用四血管阻断法建立大鼠全脑缺血再灌注损伤模型后分别于再灌注同时吸入67％N2和33％O2的混合气体、67％H2和33％O2的混合气体。各组大鼠分别于再灌注72h、5d、9d时采用免疫组化染色观察海马CA1区特异性神经元核蛋白（NeuN）染色情况及计数阳性细胞，采用比色法测定血清超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量变化。同时在再灌注9d时采用水迷宫实验检测大鼠学习记忆能力以及采用高尔基染色检测锥体神经元及树突棘数量。结果（1）免疫组化染色NeuN结果显示：与模型组相比．治疗组在再灌注72h、5d、9d时海马CA1区神经元结构相对完整且集中，形态接近正常，阳性细胞数量明显增多，差异有统计学意义（P〈0．05）。随着再灌注时间的延长，模型组再灌注各时点NeuN阳性细胞计数逐渐减少，差异有统计学意义（P〈0．05）；治疗组再灌注各时点NeuN阳性细胞计数虽有所下降，但差异无统计学意义（P〉0．05）。（2）SOD活性及MDA含量检测显示：治疗组在再灌注72h、5d、9d时血清SOD活性明显高于模型组和假手术组。而MDA含量则明显低于模型组，差异均有统计学意义（P〈0．05）。随着再灌注时间的延长，各组再灌注不同时点SOD活性相比差异无统计学意义（P〉0．05）；但模型组及治疗组MDA含量均逐渐下降，再灌注不同时点相比差异均有统计学意义（P〈0．05）。（3）在再灌注9d时，水迷宫实验发现定向航行实验中治疗组逃避潜伏期明显短于模型组，空间探索实验中第Ⅳ象限游泳时间明显长于模Objective To explore the protective effect of in-taking high concentration hydrogen gas on neurons and dendritic spines in hippocampus CA1 region of rats after globe cerebral ischemia/reperfusion （I/R） injury and its mechanism. Methods Four-vessel occlusion （4VO） was used to establish the models of global cerebral I/R injury in rats. One hundred and twenty healthy male Sprague-Dawley rats were randomly divided into 3 groups using a random number table： sham-operated group （inhaled 67% N2 and 67% O2, n=40）, model group （inhaled 67% N2 and 67% O2 during reperfusion, n=40）, and treatment group （inhaled 67% H2 and 67% O2 during reperfusion, n=40）. After 72 h, 5 and 9 d reperfusion, neuron-specific nuclear （NeuN） protein expression in the pyramidal neurons of the hippocampal CA1 region was detected with immumohistochemical staining and the positive cells were counted. And the contents of superoxide dismutase （SOD） and malondialdehyde （MDA） in sentrn were tested with colorimetry. Water maze test was used to measure the spatial orientation and memory function and Golgi staining to detect the number of dendritic spines in neurons 9 d after reperfusion. Results （1） Immunohistochemical staining of NeuN results showed that as compared with those in the model group, the neurons ofhippocampus CA1 region were significantly closer to normal with relatively intact structure, and the number of positive neurons was significantly increased in the treatment group 72 h, 5 d, and 9 d after reperfusion （P〈0.05）. With the reperfusion time being prolonged, the number of NeuN stained positive neurons at different time points of reperfusion in model group was gradually decreased （P〈0.05）, and the numeric of the NeuN stained positive neurons at different time points of reperfusion in treatment group was slightly declined without significant difference （P〉0.05）. （2） The serum SOD activity in the treatment group was significantly higher than that in the model group and sham-operated

关 键 词：氢气 脑缺血再灌注损伤 神经元 树突棘 氧化应激 

分 类 号：R741[医药卫生—神经病学与精神病学]