异丙酚诱导GAD_(67)-GFP基因敲入小鼠意识消失过程中GABA能神经元的活化  

Activation of GABAergic neurons in the GAD_(67)-GFP knock-in mouse brain during loss of consciousness induced by propofol

作　　者：周扬[1] 汪伟[2] 韩丽春[1] 高巍[1] 徐礼鲜[1] 武胜昔[2] 张惠[1] 

机构地区：[1]第四军医大学口腔医学院麻醉科 [2]第四军医大学基础部人体解剖学与组织胚胎学教研室,西安710032

出　　处：《神经解剖学杂志》2009年第4期387-392,共6页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(No.30772073;30770694)资助项目

摘　　要：探讨在系统性给予谷氨酸脱羧酶67-绿色荧光蛋白(GAD67-GFP)基因敲入小鼠异丙酚后诱导的意识消失作用中的GA-BA能神经元的活化情况。GAD67-GFP阳性小鼠20只,腹腔注射异丙酚130mg/kg(对照组腹腔注射相同体积的生理盐水)。分别在注射后5min,30min,1h进行行为学评分,随后立即处死并取脑,应用免疫组织化学观察神经元活化标记物Fos在全脑的表达分布并用免疫荧光双标的方法观察Fos与GABA能神经元的共存情况。行为学结果表明在给予异丙酚后5min和30min均能达到适度或深度麻醉效果,而1h时则处于过渡状态,行为学评分为5min时13分,30min时14分,1h时为9分。与对照组相比,Fos在海马CA1区、杏仁核、丘脑室旁核、下丘脑室旁核、梨状核、下丘脑腹内侧核、中脑导水管周围灰质的表达在三个时间点都有明显增加(P<0.05),在嗅球外丛状层有表达但与对照组相比无明显差异(P>0.05)。注射后5min、30min和1h下丘脑腹内侧核有Fos与GABA共存,有共存的细胞占该区域Fos阳性神经元的80.3%、89.7%和91.6%。下丘脑腹内侧核GABA能神经元活化是异丙酚诱导意识消失的可能机制之一。To investigate the activation of GABAergic neurons during loss of consciousness induced by propofol in the GAD67-GFP knockin mouse. Twenty GAD67-GFP knock-in mice were used in the present study. Mice in experimental group were injected propofol 130 mg/ kg （ i · p · ）, and saline was set as the negative control. Mice were graded by ethology while sacrificed at 5 rain, 30 min and 1 hr after injection, respectively. The whole brain was obtained and cut with a freezing microtome consecutively. Immunohistochemistry and double immuno-fluorescence were performed to observe the Fos expression and the coexistence of Fos and GABA in the whole brain. The scores of ethology were 13 at 5 rain, 14 at 30 min and 9 at 1h. It indicated that the mice were in the situation of moderate or deep anesthesia at 5 min and 30 min, and various transition states at 1 h. Expression of Fos protein increased remarkably in CA1, amygdaloid nuelens, thalamic paraventrieular nucleus, hypothalamic paraventricular nucleus, hypothalamie ventromedialis nucleus, periaqueduetal gray compared with the control group at three time points（ P 〈 0.05 ）. Neurons expressing Fos and GABA simultaneously could be observed in hypothalamic ventromedial nueleus at 5 rain,30 min and 1 h, the proportion of which among GFP-positive neurons was 80.3% , 89.7% and 91.6%. The activation of GABAergie neurons may be one of the meehanisms during loss of consciousness induced by systematie propofol administration.

关 键 词：异丙酚 意识消失 GABA FOS GAD67-GFP 基因敲入小鼠 

分 类 号：R96[医药卫生—药理学]