新疆医科大学学报

目的 观察模拟不同模式的睡眠剥夺对雌性大鼠脑组织的损伤作用，及其对大鼠行为学的影响。方法 以改良多平台水环境法(Modified multiple platform method, MMPM)建立不同模式的睡眠剥夺模型，将30只SPF级雌性SD大鼠分为空白对照组(n=10)、规律性睡眠剥夺组(n=10)和不规律性睡眠剥夺组(n=10)。连续实验3周，记录分析各组大鼠体重、血常规、行为学等指标变化；观察各组脑组织病理变化；采用免疫组化法检测心肌组织中钾离子通道蛋白2.1(Kv2.1)、脑组织中脑和肌肉芳香烃受体核转运样蛋白1(BMAL1)、生物钟调节蛋白(CLOCK)表达水平；采用酶联免疫吸附试验(ELISA)检测脑组织中炎症因子C-反应蛋白(CRP)和血清肿瘤坏死因子-α(TNF-α)水平。结果 睡眠剥夺后，规律性睡眠剥夺组与不规律性睡眠剥夺组大鼠体重增长幅度较小，与规律性睡眠剥夺组相比，不规律性睡眠剥夺组大鼠体重增长幅度最小，差异有统计学意义(P<0.05)。睡眠剥夺后，与空白对照组相比，规律性睡眠剥夺组WBC、LYM%水平降低，Bas%升高，差异有统计学意义(P<0.05);不规律性睡眠剥夺组WBC、MCH、LYM、LYM%水平降低的更明显，Bas%和NEU水平升高，差异有统计学意义(P<0.05);与规律性睡眠剥夺组相比，不规律性睡眠剥夺组MCH、LYM水平降低，NEU水平升高，差异有统计学意义(P<0.05)。睡眠剥夺前，3组大鼠站立次数、修饰次数无显著差异，睡眠剥夺后，与空白对照组相比，规律性睡眠剥夺组大鼠的站立次数及修饰次数增加(P<0.05);与规律性睡眠剥夺组相比，不规律性睡眠剥夺组大鼠的站立次数及修饰次数降低(P<0.05)。与空白对照组及规律性睡眠剥夺组大鼠相比，不规律性睡眠剥夺组大鼠海马CAI区出现病理损伤。与空白对照组相比，规律性睡眠剥夺组大鼠心肌组织中Kv2.1、脑组织中CLOCK表达量均降低(P<0.05),不规律性睡眠剥夺组大鼠心肌组织中Kv2.1、脑组织中BMAL1、CLOCK表达降低的更明显，差异均具有统计学意义(P<0.05)。与空白对照组及规律性睡眠剥夺组大鼠比较，不规律性睡眠剥夺组大鼠脑组织中CRP及TNF-α水平均升高，差异有统计学意义(P<0.05)。结论 通过模拟不同模式的睡眠剥夺，模型大鼠在不规律性睡眠剥夺模式下，脑组织损伤与焦虑情绪逐渐增加，可能与生物钟紊乱、炎症反应、海马体神经细胞受损等多种因素有关。

Objective To observe the damage effect of simulated different modes of sleep deprivation on the brain tissue of female rats, and to explore its effect on the behavior of rats. Methods A sleep deprivation model was established by modified multi-platform water environment(MMPM) and 30 SPF female SD rats were divided into blank control group(n=10), regular sleep deprivation group(n=10) and irregular sleep deprivation group(n=10). The changes of body weight, blood routine and behavior of the rats in each group were recorded and analyzed for 3 weeks.The pathological changes of brain tissue in each group were observed. Immunohistochemistry was used to detect the expression levels of Kv2.1 in myocardial tissue, BMAL1 and CLOCK in brain tissue. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory cytokines CRP and TNF-α in brain tissues. Results After sleep deprivation, the weight gain of of rats in the regular sleep deprivation group and the irregular sleep deprivation group was smaller. Compared with the regular sleep deprivation group, the weight gain of rats in the irregular sleep deprivation group was the smallest, and the difference was statistically significant(P<0.05). After sleep deprivation, compared with the blank control group, the levels of WBC and LYM% in the regular sleep deprivation group decreased, and the Bas% increased, with statistically significant differences(P<0.05); in the irregular sleep deprivation group, the levels of WBC, MCH, LYM and LYM% decreased more significantly, and the Bas% and NEU levels increased, with statistically significant differences(P<0.05); compared with the regular sleep deprivation group, the levels of MCH and LYM in the irregular sleep deprivation group decreased, and the NEU level increased(P<0.05). Before sleep deprivation, there were no significant differences in the standing times and modification times among the groups. After sleep deprivation, compared with the blank control group, the standing times and modification times of rats in the regular sleep deprivation group increased significantly(P<0.05). Compared with the regular sleep deprivation group, the standing times and modification times of rats in the irregular sleep deprivation group decreased(P<0.05). Compared with the blank control group and the regular sleep deprivation group, pathological damage occurred in the hippocampal CA1 area of rats in the irregular sleep deprivation group. Compared with the blank control group, the expression levels of Kv2.1 in the myocardial tissue and CLOCK in the brain tissue of rats in the regular sleep deprivation group decreased(P<0.05), and the expression levels of Kv2.1 in the myocardial tissue, BMAL1 and CLOCK in the brain tissue of rats in the irregular sleep deprivation group decreased more significantly, and all the differences were statistically significant(P<0.05). Compared with the blank control group and the regular sleep deprivation group, the levels of CRP and TNF-α in the brain tissues of rats in the irregular sleep deprivation group increased, and the differences were statistically significant(P<0.05). Conclusion By simulating different modes of sleep deprivation, brain tissue damage and anxiety in model rats under irregular sleep deprivation patterns were gradually increased, which may be caused by various factors such as circadian clock disorder, inflammatory response and damage to neurons in the hippocampus.