睡眠剥夺引起成年大鼠脑海马神经细胞增殖及干细胞因子的表达

Changes of the expression of stem cell factor and nerve cell proliferation in hippocampus of rats induced by sleep deprivation

目的:观察快眼动睡眠剥夺对成年大鼠脑海马齿状回神经细胞增殖的影响,并探讨干细胞因子mRNA表达的变化。方法:实验于2000-03/2002-03在郑州大学医学院完成。将SD大鼠36只随机分为正常对照组、快眼动睡眠剥夺组及快眼动睡眠剥夺对照组,每组12只,其中6只动物在睡眠剥夺后立即灌流固定,余6只动物饲养3周后进行同样的处理。各组动物于剥夺睡眠当日12:00腹腔注射5-溴-2-脱氧尿嘧啶核苷100mg/kg,连续剥夺快眼动睡眠72h,应用细胞增殖标记免疫组化及原位杂交染色观察各组动物脑海马神经细胞增殖分化及干细胞因子mRNA表达的变化。结果:36只大鼠全部进入结果分析。①快眼动睡眠剥夺组脑海马齿状回区细胞增殖率[(22.21±0.84)%]明显高于快眼动睡眠剥夺对照组和正常对照组[(3.97±0.53)%和(3.81±0.50)%,(F=1594,P<0.001)]。正常对照组和快眼动睡眠剥夺对照组脑海马齿状回区细胞增殖率相似(P>0.05)。②快眼动睡眠剥夺组脑海马齿状回区干细胞因子mRNA表达增强,阳性积分高于快眼动睡眠剥夺对照组和正常对照组[(134.50±52.06),(67.69±16.95),(70.61±19.84)分,(F=6.305,P<0.05)],睡眠剥夺3周后部分增殖细胞可表达成熟神经元标记神经元特异性烯醇化酶。

AIM:To study the effect of rapid eye movement sleep deprivation on nerve cell proliferation in dentate gyrus of hippocampus in adult rats, and study the expre ssion of stem cell factor mRNA. METHODS:The experiment was finished in Zhengzhou University from March 2000 to March 2002.A total 36 SD rats were divided randomly into normal control group, rapid eye movement sleep deprivation group and rapid eye movement sleep deprivat ion control group,12 rats in each group,6 of which received perfusion following sleep deprivation, and the others received the same treatment after 3 week feed .Rats were injected with 100 mg/kg 5 bromo 2 deoxyuridine into abdominal cavi ty at 12:00 on the day of sleep deprivation.Double labeling immunohistochemistry and in situ hybridization staining were used to observe the nerve cell prolifer ation and the expression of stem cell factor mRNA. RESULTS: According to intention to treat, all rats were involved in the resu lt analysis.①The proliferation rate in dentate gyrus of hippocampus increased i n rapid eye movement sleep deprivation group [(2.21±0.84)]%as compared with th at in rapid eye movement sleep deprivation control group and normal control grou p [(3.97±0.53)%, (3.81±0.50)%,F=1 594,P< 0.001],and there was no significant difference between the later two groups(P >0.05);②The positive expression of s tem cell factor mRNA in dentate gyrus of hippocampus increased in rapid eye move ment sleep deprivation group as compared with that in rapid eye movement sleep d eprivation control group and normal control group (134.50±52.06,67.69±16.95,70 .61±19.84, F=6.305,P< 0.05).After 3 week sleep deprivation,some proliferate ce lls could express mature neuron marker neuron specific enolase. CONCLUSION: Rapid eye movement sleep deprivation can increase nerve cell proli feration in dentate gyrus of hippocampus in adult rats.After 3 week sleep depri vation, some proliferate cells can express mature neuron marker neuron specifi c enolase.The result indicates that sleeping not only affects the maturity and d evelopment of nervous system, but also plays an important role in nerve cell pro liferation in brain.