摘要
目的:观察异氟醚对氧糖剥夺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,造成氧糖剥夺离体神经元缺血模型,4h后复氧、复糖,模拟再灌注模型。随机分为3组:C组为对照组;A组用1MAC异氟醚通入培养液预处理1h;B组用2MAC异氟醚通入培养液预处理1h。各组分别于复氧复糖后4、12和24h取神经元细胞计数后作四唑盐(MTT)比色试验。并取复氧、复糖24h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,造成氧糖剥夺离体神经元缺血模型30min后分成两组后分别按C组和A组处理,复氧、复糖4h后,免疫组化SABC法染色,比较nNOS和iNOS活性。结果:复氧、复糖后A组和B组MTT各时点值均大于C组,且台盼蓝染色阳性细胞数百分比A组和B组均小于C组,但A组和B组间无显著差异。免疫组化染色结果显示:A组nNOS和iNOS的表达灰度值均低于C组。结论:异氟醚能减轻氧糖剥夺离体神经元的损伤,抑制神经元nNOS和iNOS活性是其可能机制之一。
Objective：To investigate the protective effect of isoflurane on the oxygen and glucose deprivation（OGD） in cultured neuronal cell. Methods： The cultured neurons dissociated from the mouse brain were observed on the 14th day after cultured. To make the ischemia-reperfusion model in vitro, the neurons were exposed to OGD 4h before they were placed to nomal culture media. Group Control was taken as blank group. In group A, 1 MAC isoflurane was given to neurons in the culture media before the oxygen-glucose（OG）,and 2 MAC was given in Group B. The neuronal injury was detected by MTT assay in different time （4h,12h,24h after OG regain） and by trypan blue stained. Furthermore, the activity expression of nNOS and iNOS were detected by immunohistochemical technique between Group C and Group A. Results： The activity of neurons （MTT assay） was larger in Group A and Group B than that in Group C. The percentage of postive neurons stained by trypan blue was samaller in Group A and Group B than that in Group C. There was no significant difference between Group A and Group B. In view of the positive expression of nNOS and iNOS, there was significant difference between Group C and Group A. Conclusion： Isoflurane can mitigate the damage of OGD neurons and the restrained effect on nNOS and iNOS may be a part of the mechanism.
出处
《中国临床医学》
2009年第2期291-293,共3页
Chinese Journal of Clinical Medicine
关键词
异氟醚
神经元
氧糖剥夺
Isofurane
Neuron
Oxygen and glucose deprivation
作者简介
通讯作者 李颖川，E-mail：yingchli@yahoo．com．cn
