摘要
目的研究不同时间快速动眼(REM)睡眠剥夺对大鼠皮质、海马及延髓葡萄糖调控蛋白(glucose-regulated protein,GRP)表达的影响。方法大鼠随机分为睡眠剥夺1、3、5、7d(SD 1 d,SD 3d,SO 5d,SD 7d,n=10)组、剥夺7d后恢复睡眠6、12h(SD 7d/RS 6h,SD 7d/RS 12h,n= 10)组、环境对照(TC,n=10)组和空白对照(CC,n=10)组。采用改良多平台睡眠剥夺法进行不同时间REM睡眠剥夺。应用免疫组织化学方法及半定量逆转录-聚合酶链反应(RT-PCR)技术检测脑组织标本中GRP78、GRP94表达分布规律及时空的变化。结果睡眠剥夺d1皮质、海马区GRP78、GRP94蛋白表达增多(与TC、CC组比较,P<0.05),d 3达高峰,d 5、7与TC、CC组比较无显著差异(P>0.05);延髓区无明显改变;恢复睡眠后只有延髓区GRP94表达增加。RT-PCR显示睡眠剥夺l d GRP78转录增加,d 3达峰值,d 5、7转录无增加;GRP94转录在剥夺期间无增加;恢复睡眠后GRP78、GRP94 mRNA增加。睡眠剥夺d 3 GRP蛋白表达改变皮质区最为明显(P<0.01),其次为海马区(P<0.05)。结论短时间睡眠剥夺后GRP78、GRP94表达渐升高,可能是内质网启动了自身稳定调节的保护功能;长时间睡眠剥夺导致内质网功能障碍,GRP78、GRP94表达下降。皮质区对睡眠剥夺引起内质网应激的保护性反应最为强烈。
AIM To observe the influence of rapid eye movement （REM） sleep deprivation on expressions of the glucose-regulated protein, GRP78 and GRP94 in the rats cerebral cortex, hippocampus and medulla oblongata on different sleep deprivation stages. METHODS Eighty Wistar rats were distributed randomly into sleep deprivation （SD, n = 40） group, revival sleep （RS, n = 20） group, tank control （TC, n = 10） group and cage control （CC, n = 10） group. The SD group was divided into 4 subgroups, including the rats＇ sleep deprivation for 1, 3, 5 d and 7 d separately. The RS group was divided into two subgroup, including revival sleep of 6 h and 12 h. The modified multiple platform methods （MMPM） were established in the REM sleepdeprived rats at different time. The Streptavdin-peroxidase-biotin （SP） immunohistochemistry methods and RTPCR techniques were used to detect the distribution system and time and space changes of GRP78 and GRP94 expressions in the rat frontal cortex, hippocampus and medulla oblongata. RESULTS Immunohistochemistry staining results showed that compared with those of the rats in TC and CC, the expressions of GRP78 and GRP94 protein increased on 1 d after SD in the cortex and hippocampus region （P 〈 0.05）, and reached the highest on d 3 after SD. While on d 5 and d 7 after SD, the expressions of GRP78 and GRP94 were not different from those of TC and CC （P 〉 0.05）. The expressions of GRP78 and GRP94 protein have no obvious changes in the medulla region. Only expressions of GRP94 increased in the medulla after revival sleep. RT-PCR showed that the transcription of GRP78 increased after one 1 d after SD, and reached the highest on d 3, and no improvement on d 5 and d 7 after SD. The transcription of GRP94 had no improvement during SD. The transcription of GRP78 and GRP94 increased after revival sleep. The changes of the GRP proteins expressions on d 3 were the most obvious in the cerebral cortex region （P 〈 0.01 ） , then in the hippocampus region （P 〈 0.05）. C
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2008年第4期249-253,共5页
Chinese Journal of New Drugs and Clinical Remedies
基金
上海市科技发展基金(024119029)
中国人民解放军第二军医大学附属长征医院“十一五”、“三重二优”学科人才建设基金(2005312)
关键词
睡眠剥夺
海马
延髓
葡萄糖调控蛋白
大脑皮质
sleep deprivation
hippocampus
medulla oblongata
glucose-regulated proteins
cerebralcortex
