快速眼动期睡眠剥夺对大鼠海马齿状回神经元增殖与凋亡的影响  

Effects of rapid eye movement sleep deprivation on proliferation and apoptosis of hippocampal dentate gyrus neurons in rats

作　　者：梁小敏[1] 陈甲[1] 许志强[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所神经内科,重庆400042

出　　处：《第三军医大学学报》2013年第19期2014-2018,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81271411);重庆市自然科学基金面上项目(CSTC2012jjA10098)~~

摘　　要：目的探讨快速眼动期(rapid eye movement,REM)睡眠剥夺(sleep deprivation,SD)对大鼠海马齿状回神经元增殖与凋亡的影响。方法 45只Sprague-Dawley大鼠按随机数字表法分为3组:①普通鼠笼对照组(CC,n=15);②睡眠剥夺组(SD,n=15);③睡眠恢复组(RS,n=15)。采用改良多平台睡眠剥夺法连续72 h剥夺大鼠REM睡眠后,利用BrdU标记增殖细胞,运用免疫组织化学方法观察大鼠海马齿状回神经元增殖情况,并利用免疫荧光染色检测大鼠海马齿状回新生神经元标记物微管相关蛋白(doublecortin,DCX)和星形胶质细胞标记物胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的表达情况。采用TUNEL法观察大鼠海马神经元凋亡情况。结果免疫组化结果显示SD组及RS组每张切片BrdU阳性细胞平均数均明显少于CC组(P<0.01),但是RS组与SD组相比无显著性差异(P>0.05)。免疫荧光染色结果显示,DCX阳性细胞在BrdU阳性细胞中所占比例最大,各组间BrdU/DCX、BrdU/GFAP双阳性细胞的百分比相差不显著(P>0.05)。CC组、SD组及RS组各组海马区均未见TUNEL阳性细胞。结论 REM睡眠剥夺抑制海马齿状回神经元增殖,短期睡眠恢复后无显著改善。REM睡眠剥夺对海马齿状回神经元的分化和凋亡无影响。Objective To determine the effect of rapid eye movement （REM） sleep deprivation on the proliferation and apoptosis of hippocampal dentate gyrus neurons in rats. Methods Forty-five Sprague- Dawley rats were randomly divided into 3 groups： cage control group （CC）, sleep deprivation group （SD） and sleep revival group （RS）. The modified multiple platform method was used to establish sleep deprivation model for 72-hour REM sleep deprivation in rats. Immunohistoehemieal assay was used to observe the neuron prolifera- tion and differentiation in the dentate gyms of rats, while the apoptosis of hippoeampal neuron was detected by TUNEL assay. Immunofluorescenee staining was used to detect the expression of newborn neurons＇ biomarker, microtubule-associated protein, doublecortin （DCX） , and astrocytes＇ biomarker, glial fibrillary acidic protein （GFAP） in the hippocampal dentate gyms. Results The average numbers of BrdU positive cells were signifi- cantly lower in SD and RS groups than in CC group （ P 〈 0.01 ） , but there was no significant difference between the former 2 groups. Immunofluoreseence staining showed that the majority of BrdU positive cells were DCX positive cells. There was no significant difference in the phenotype of BrdU positive cells （BrdU/DCX and BrdU/GFAP） between each group. No TUNEL positive cells were seen in the hippocampus of all groups. Conclusion REM sleep deprivation inhibits the neuron proliferation in the dentate gyrus of rats, and there is no significant improvement after short-term sleep revival. However, the sleep deprivation has no effect on the differentiation and apoptosis in hippoeampal neurons.

关 键 词：快速眼动期睡眠剥夺 海马齿状回 神经元增殖 凋亡 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R338.63[医药卫生—基础医学]