连续睡眠剥夺对大鼠肝生物钟基因及肝损伤标志物表达水平的影响  

Effect of continuous sleep deprivation on liver circadian clock gene expression and alteration of liver bio-marker levels in rats

作　　者：顾晔[1,2] 邢陈[2] 宋伦[2] 黎卫平[1] 

机构地区：[1]兰州大学基础医学院病理学研究室,兰州730000 [2]军事医学科学院基础医学研究所应激医学研究室,北京100850

出　　处：《军事医学》2017年第3期174-177,共4页Military Medical Sciences

基　　金：军队"十二五"重大资助项目(AWS14J011);北京市自然科学基金青年基金资助项目(7164287);细胞应激生物学国家重点实验室(厦门大学)开放基金资助项目(SKLCSB2016KF010)

摘　　要：目的探讨36 h连续睡眠剥夺(SD)对大鼠肝、肾生物钟基因表达水平及大鼠尿液中肝损伤标志物总胆汁酸(TBA)含量的影响。方法将12只大鼠随机分为正常对照组、睡眠剥夺组(SD组)。SD组大鼠使用睡眠剥夺仪进行睡眠剥夺,连续36 h后,暴露腹腔取肝、肾,用RT-PCR方法检测大鼠肝、肾组织中生物钟基因转录表达水平的变化情况,并对生物钟基因的蛋白表达产物进行免疫印迹分析;暴露盆腔取尿液,用ELISA试剂盒检测大鼠尿液中肝损伤标志物TBA含量的变化情况。结果与对照组相比,SD组大鼠肝生物钟基因clock,bmal1的mRNA表达水平明显下调(P<0.05),肾同种生物钟基因表达水平在睡眠剥夺前后无明显变化。肝生物钟基因的蛋白表达水平与转录水平一致。同时,SD组大鼠与对照组大鼠相比尿液中TBA含量明显上调(P<0.001)。结论 36 h连续睡眠剥夺可导致大鼠肝生物钟基因表达和胆固醇代谢紊乱,TBA可作为昼夜节律紊乱致肝损伤效应的无创性生物标志物。Objective To investigate the effect of 36 h continuous sleep deprivation（ SD） on circadian clock gene expression in the rat liver and kidney and the alteration of urine biomarker levels. Methods Twelve rats were randomly divided into control group and SD group. An SD device was used to deprive the rats of sleep. After 36 h continuous SD,the abdominal cavity was exposed to obtain livers and kidneys,and RT-PCR and Western blotting were used to detect expression of clock genes. Then,the pelvic cavity was exposed to obtain urine,and the changes in bio-marker total bile acids（ TBA） were tested with ELISA. Results Compared with the control group,the mRNA level of liver clock and bmal1 was obviously reduced in the SD-treated rats（ P〈0. 05）. However,no obvious change was found in the samples from the kidney. Sharp down-regulation of CLOCK and BMAL1 protein expression was also observed in the rat liver after SD treatment. Urine TBA content in SD treated rats was raised obviously（ P 0. 001）,compared with control. Conclusion Thirty-six hours of continuous SD could result in deregulation of circadian clock gene and cholesterol metabolism disorder in the rat liver. TBA might be used as a noninvasive biomarker of liver injury under SD stress conditions.

关 键 词：连续睡眠剥夺 生物钟基因 昼夜节律 肝损伤标志物 

分 类 号：R575[医药卫生—消化系统]