睡眠剥夺对幼鼠小脑沉默信息调节因子6表达的影响  

Effect of sleep deprivation on expression of SIRT6 in cerebellum of immature mice

作　　者：肖岚 高尘怡 陈婷[1] 张学敏 钟琦[1] 曹玥 唐李娟 陈畅[1] 张宗泽[1] Xiao Lan;Gao Chenyi;Chen Ting;Zhang Xuemin;Zhong Qi;Cao Yue;Tang Lijuan;Chen Chang;Zhang Zongze(Department of Anesthesiology,Zhongnan Hospital of Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院麻醉科,武汉430071

出　　处：《中华麻醉学杂志》2024年第1期41-45,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81901109,82371290);湖北省中央引导专项(2022BGE236)。

摘　　要：目的评价睡眠剥夺对幼鼠小脑沉默信息调节因子6(SIRT6)表达的影响。方法SPF级健康雄性C57BL/6小鼠50只,4周龄,体质量14~16 g,采用随机数字表法分为2组(n=25):对照组(Con组)和睡眠剥夺组(SD组)。采用多平台水环境法制备小鼠睡眠剥夺模型,每天睡眠剥夺20 h,连续10 d。睡眠剥夺后行平衡木实验检测小鼠平衡和协调能力,麻醉后处死小鼠,取小脑组织,透射电子显微镜观察超微结构,采用高尔基染色法测定小脑浦肯野细胞树突棘密度,采用免疫荧光法检测小脑浦肯野细胞SIRT6和钙结合蛋白D-28k(CbD-28k)共表达情况,检测小脑葡萄糖转运体3(Glut3)表达。结果与Con组比较,SD组小鼠平衡木通过时间延长,后足滑脱次数增加,小脑4-6cb神经元突触间隙增大,突触后致密物厚度、浦肯野细胞树突棘密度及SIRT6与CbD-28k共表达阳性细胞数降低,Glut3表达下调(P<0.05)。结论睡眠剥夺降低幼鼠平衡和协调能力的机制与下调小脑浦肯野细胞SIRT6表达,降低神经元糖代谢,从而损伤小脑突触可塑性有关。Objective To evaluate the effect of sleep deprivation on the expression of sirtuin 6(SIRT6)in the cerebellum of immature mice.Methods Fifty SPF healthy male C57BL/6 mice,aged 4 weeks,weighing 14-16 g,were divided into 2 groups(n=25 each)using a random number table method:control group(Con group)and sleep deprivation group(SD group).The chronic sleep deprivation model was prepared by using the multi-platform water environment method,with 20 h of sleep deprivation per day for 10 consecutive days.After sleep deprivation,a balance beam experiment was performed to test the balance and coordination ability of mice.The mice were sacrificed after anesthesia and cerebellar lobular IV-VI(4-6 cb)tissues were taken for microscopic examination of the ultrastructure(with a transmission electron microscope)and for determination of the dendritic spine density of cerebellar 4-6cb Purkinje neurons(by Golgi staining),co-expression of SIRT6 and Calbindin D-28k(CbD-28k)and expression of glucose transporter Glut3 of cerebellar 4-6cb(by immunofluorescence staining).Results Compared with group Con,the duration of passage through the balance beam was significantly prolonged,and the number of posterior foot slips was increased,the synaptic gap of cerebellar 4-6cb neurons was increased,the thickness of postsynaptic density was increased,the density of dendritic spines of Purkinje cells and the number of positive cells co-expressing SIRT6 and CbD-28k were decreased,and the expression of Glut3 was down-regulated in group SD(P<0.05).Conclusions The mechanism by which sleep deprivation decreases the abilities of balance and coordination is related to down-regulating SIRT6 expression in cerebellar Purkinje cells and decreasing neuronal glucose metabolism,thus damaging the synaptic plasticity of cerebellum in immature mice.

关 键 词：睡眠剥夺 小脑 沉默信息调节因子6 

分 类 号：R614[医药卫生—麻醉学]