高氧液对氧糖剥夺离体神经元损伤的保护作用  

The protective effects of hyperoxia solution on the oxygen and glucose deprivation in cultured neuronal cells

作　　者：曾真[1] 张惠[1] 徐礼鲜[1] 

机构地区：[1]第四军医大学口腔医学院麻醉科,西安市710032

出　　处：《临床麻醉学杂志》2004年第6期353-355,共3页Journal of Clinical Anesthesiology

基　　金：国家科技部创新基金项目 ( 0 0c2 62 2 6110 464 ) ;陕西省自然科学基金项目 ( 99sm3 9)

摘　　要：目的　观察高氧液对氧糖剥夺 (OGD)离体神经元损伤的保护作用。方法　新生小鼠神经元原代培养 14d后 ,造成OGD离体神经元缺血模型 ,4h后复氧复糖 (ROG) ,模拟再灌注模型。随机分组 :对照组 (A组 ,n =6 ) ;治疗组在ROG时 ,根据剂量的不同分为两组 :B组 (n =6 )高氧液浓度为 33 3% ,C组 (n =6 )高氧液浓度为 2 0 0 %。各组分别于ROG后 4、12、2 4h取神经元细胞记数后行四唑盐 (MTT)比色试验。并取ROG 2 4h各组神经元细胞做台盼蓝染色 ,计阳性细胞数百分比。另取原代培养神经元 ,造成OGD离体神经元缺血模型 30min后 ,分A组 (n =10 )和B组 (n =10 )ROG 4h后 ,免疫组化SABC法染色 ,比较Ⅰ型 氧化氮合酶 (nNOS)和Ⅱ型 氧化氮合酶 (iNOS)活性。结果　ROG后 ,B组和C组MTT各时点吸光值 (OD)均高于A组 ,且台盼蓝染色阳性细胞数百分比B组和C组均低于A组 ,但B组和C组间差异不显著。免疫组化染色结果显示 ,B组nNOS和iNOS的表达灰度值均高于A组。结论　高氧液能减轻OGD离体神经元损伤 ,抑制神经元nNOS和iNOS活性是其可能机制之一。Objective To investigate the protective effects of hyperoxia solution(HS) on the oxygen and glucose deprivation(OGD) in cultured neuronal cells.Methods The cultured neurons dissociated from mouse brain were observed on the 14th day after culture.To make the ischemia-reperfusion model in vitro,the neurons were exposed to OGD 4 h before they were placed into normal culture media.Group A was taken as blank control.In group B,the HS which took half volume of the culture media was given to neurons after the oxygen-glucose (OG) regained.HS took 25 percent volume of the culture media in group C.The neuronal injury was detected by MTT assay in different time points (4,12,24 h after OG regain) and by trypan blue stain.Furthermore,the activity expression of nNOS and iNOS was compared by immunohistochemical technique between group A and group B. Results MTT assay was larger in group B and group C than that in group A.The percentage of positive neurons stained by trypan blue was smaller in group B and group C than that in group A,which was not significantly different between group B and group C.The positive expression of nNOS and iNOS was significantly different between group A and group B. Conclusion Hyperoxia solution can mitigate the damage of OGD neurons.The depressive effects of HS on nNOS and iNOS may be a part of the mechanisms.

关 键 词：高氧液 氧糖剥夺 神经元损伤 保护作用 神经元缺血 再灌注损伤 

分 类 号：R965[医药卫生—药理学]