睡眠剥夺对大鼠血脑屏障的影响

doi:10.3969/j.issn.1006-9771.2019.09.006

《中国康复理论与实践》 ›› 2019, Vol. 25 ›› Issue (9): 1020-1025.doi: 10.3969/j.issn.1006-9771.2019.09.006

睡眠剥夺对大鼠血脑屏障的影响

李丹丹^1,2, 马芮^1,3, 黄敏莹⁴, 赵弘轶¹, 黄勇华¹

1.中国人民解放军总医院第七医学中心神经内科，北京市 100700
2.山西医科大学第二临床医学院，山西太原市 030001
3.安徽医科大学，安徽合肥市 230032
4.南方医科大学第二临床医学院，广东广州市 510515

收稿日期:2019-01-17 修回日期:2019-03-05 出版日期:2019-09-25 发布日期:2019-09-25
通讯作者: 黄勇华，E-mail: huangyh@163.com E-mail:huangyh@163.com
作者简介:李丹丹(1988-)，女，汉族，山东济南市人，硕士研究生，医师，主要研究方向：脑小血管病及生命早期应激。
基金资助:
吴阶平医学基金会项目(No. 320.6750.18456)

Influence of Sleep Deprivation on Blood-brain Barrier in Rats

LI Dan-dan^1,2, MA Rui^1,3, HUANG Min-ying⁴, ZHAO Hong-yi¹, HUANG Yong-hua¹

1.Department of Neurology, the Seventh Medical Center of PLA General Hospital, Beijing 100700, China
2.The Second Clinical Medical College of Shanxi Medical University, Taiyuan, Shanxi 030001, China
3.Anhui Medical University, Hefei, Anhui 230032, China
4.The Second Clinical Medical College of Southern Medical University, Guangzhou, Guangdong 510515, China

Received:2019-01-17 Revised:2019-03-05 Published:2019-09-25 Online:2019-09-25
Contact: HUANG Yong-hua, E-mail: huangyh@163.com E-mail:huangyh@163.com
Supported by:
Supported by Wu Jieping Medical Foundation (No. 320.6750.18456)

摘要/Abstract

摘要： 目的探讨睡眠剥夺(SD)对成年雄性大鼠血脑屏障的影响。方法健康雄性Sprague-Dawley大鼠90只随机等分为SD组、SD恢复(SDR)组和对照(K)组。SD组、SDR组大鼠采用水平台法连续SD 5 d，SDR组在SD后再正常饲养2 d。K组不采取措施。Evens Blue (EB)心脏灌注观察大脑染液渗漏情况，Western blotting检测皮质下组织zonula occludens-1 (ZO-1)、Occluding、Claudin-5、Bax/BCL-2、P53、caspase-3含量，免疫荧光染色观察大脑皮质内皮细胞、神经元细胞及其凋亡情况。结果 SD组多个脑叶广泛皮质可见EB染液渗出；SDR组相应部位也有染液渗出，但较轻微；K组未见染液渗出。P53、Caspase-3、Bax/BCL-2表达SD组最多，K组最少；ZO-1、Occludin、Claudin-5表达SD组最少，K组最多；SDR组均介于两者之间(F > 39.915, P < 0.001)。皮质中CD₃₁、NeuN阳性细胞数SD组最少，K组最多；TUNEL阳性细胞SD组最多，K组最少；SDR组介于两者之间(F > 142.056, P < 0.001)。结论 SD能导致大鼠血脑屏障通透性增加，紧密连接蛋白表达降低，神经元凋亡增加，皮质神经元和内皮细胞数目减少；睡眠恢复可部分缓解其对血脑屏障的影响。

关键词: 睡眠剥夺, 血脑屏障, 紧密连接蛋白, 细胞凋亡, 睡眠恢复, 大鼠

Abstract: Objective To investigate the influence of sleep deprivation (SD) on blood-brain barrier in adult male rats. Methods A total of 90 healthy male Sprague-Dawley rats were randomly divided into SD group, SD and recovery (SDR) group and control (K) group. SD group and SDR group were continuously deprived of sleep for five days by horizontal table, and then, SDR group were fed normally for two days after SD. K group accepted no intervention. The leakage of Evens Blue (EB) in brain was observed after EB perfusion. The expression of zonula occludens-1 (ZO-1), Occluding, Claudin-5, Bax/BCL-2, P53 and caspase-3 in the cortex was detected with Western blotting. The apoptosis of neurons and endothelial cells in cortex was observed with immunofluorescence staining. Results In SD group, EB was observed in multiple cerebral lobe and extensive cortex, and it was also observed in SDR group in a milder way, but not observed in K group. The expression of P53, Caspase-3, Bax/BCL-2 in the cerebral cortex was the most in SD group, and the least in K group; while the expression of ZO-1, Occluding and Claudin-5 was the least in SD group, and the most in K group, and it was in-between in SDR group (F > 39.915, P < 0.001). The CD₃₁ and NeuN positive cells decreased in cortex were the least in SD group, and the most in K group, while the TUNEL positive cells were the most in SD group, and the least in K group, and the levels of CD₃₁, NeuN and TUNEL positive cells were in-between in SDR group (F > 142.056, P < 0.001). Conclusion SD may lead to dysfunction of permeability of blood-brain barrier, while decrease expression of tight junction protein, and increase apoptosis of neurons in rats to reduce the neurons and endothelial cells in cerebral cortex. Sleep recovery may partly alleviate these impacts.

Key words: sleep deprivation, blood-brain barrier, tight junction protein, apoptosis, sleep recovery, rats

中图分类号:

R749.7

李丹丹, 马芮, 黄敏莹, 赵弘轶, 黄勇华. 睡眠剥夺对大鼠血脑屏障的影响[J]. 《中国康复理论与实践》, 2019, 25(9): 1020-1025.

LI Dan-dan, MA Rui, HUANG Min-ying, ZHAO Hong-yi, HUANG Yong-hua. Influence of Sleep Deprivation on Blood-brain Barrier in Rats[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2019, 25(9): 1020-1025.

参考文献

1 ObermeierB, VermaA, RansohoffR M. The blood-brain barrier [J]. Handb Clin Neurol, 2016, 133: 39-59.
2 DanemanR, PratA. The blood-brain barrier [J]. Cold Spring Harb Perspect Biol, 2015, 7(1): 32-34.
3 SharifY, JumahF, CoplanL, et al. Blood brain barrier: a review of its anatomy and physiology in health and disease [J]. Clin Anat, 2018, 31(6): 812-823.
4 KeaneyJ, CampbellM. The dynamic blood-brain barrier [J]. FEBS J, 2015, 282(21): 4067-4079.
5 ChengS, GaoW W, XuX, et al. Methylprednisolone sodium succinate reduces BBB disruption and inflammation in a model mouse of intracranial haemorrhage [J]. Brain Res Bull, 2016, 127: 226-233.
6 WangH, ChenF, DuY F, et al. Targeted inhibition of RAGE reduces amyloid-β influx across the blood-brain barrier and improves cognitive deficits in, db/db, mice [J]. Neuropharmacology, 2018, 131: 143-153.
7 ShiX, BaiY, ZhangG, et al. Effects of over-expression of SOD2 in bone marrow-derived mesenchymal stem cells on traumatic brain injury [J]. Cell Tissue Res, 2018, 372(1): 67-75.
8 ChoiK H, KimH S, ParkM S, et al. Regulation of caveolin-1 expression determines early brain edema after experimental focal cerebral ischemia [J]. Stroke, 2016, 47(5): 1336-1343.
9 MoonJ H, NaJ Y, LeeM C, et al. Neuroprotective effects of systemic cerebral endothelial cell transplantation in a rat model of cerebral ischemia [J]. Am J Transl Res, 2016, 8(5): 2343-2353.
10 PulakL M, JensenL. Sleep in the intensive care unit: a review [J]. J Intensive Care Med, 2016, 31(1): 14-23.
11 赵忠新. 睡眠医学[M]. 北京:人民卫生出版社, 2016: 83-87.
12 吴洪敏,娄培安,陈培培,等. 睡眠时间与高血压、糖尿病和冠心病关系的现况研究[J]. 中华全科医学, 2013, 11(9): 1438-1439.
13 OkajimaI, ChenJ. The effect of insomnia on changes in anxiety, depression, and social function after a transdiagnostic treatment targeting excessive worry [J]. Sleep Biol Rhythms, 2017, 15(3): 243-249.
14 GoelN. Neurobehavioral effects and biomarkers of sleep loss in healthy adults [J]. Curr Neurol Neurosci Rep, 2017, 17(11): 89.
15 HeJ, HsuchouH, HeY, et al. Sleep restriction impairs blood-brain barrier function [J]. J Neurosci, 2014, 34(44): 14697-14706.
16 PanW, KastinA J. The Blood-brain barrier: regulatory roles in wakefulness and sleep [J]. Neuroscientist, 2017, 23(2): 124-136.
17 SharmaA, MuresanuD F, LafuenteJ V, et al. Sleep deprivation-induced blood-brain barrier breakdown and brain dysfunction are exacerbated by size-related exposure to Ag and Cu nanoparticles. Neuroprotective effects of a 5-HT3 receptor antagonist ondansetron [J]. Mol Neurobiol, 2015, 52(2): 867-881.
18 TobaldiniE, CostantinoG, SolbiatiM, et al. Sleep, sleep deprivation, autonomic nervous system and cardiovascular diseases [J]. Neurosci Biobehav Rev, 2017, 74(Pt B): 321-329.
19 GangwischJ E. A review of evidence for the link between sleep duration and hypertension [J]. Am J Hypertens, 2014, 27(10): 1235-1242.
20 AlabriM A, JajuD, AlsinaniS, et al. Habitual sleep deprivation is associated with type 2 diabetes: a case-control study [J]. Oman Med J, 2016, 31(6): 399-403.
21 MillerN L, ShattuckL G, MatsangasP. Sleep and fatigue issues in continuous operations: a survey of U.S. Army officers [J]. Behav Sleep Med, 2011, 9(1): 53-65.
22 HeatonK J, MauleA L, MarutaJ, et al. Attention and visual tracking degradation during acute sleep deprivation in a military sample [J]. Aviat Space Environ Med, 2014, 85(5): 497-503.
23 EspositoM J, OcchioneroM, CicognaP. Sleep deprivation and time-based prospective memory [J]. Sleep, 2015, 38(11): 1823-1826.
24 HakimehS, VahidS. Effects of exercise and/or sleep deprivation on anxiety-like behavior and body weight of female rats [J]. Asian J Psychiatr, 2017, 28: 26-27.
25 SayginM, OzgunerM F, OnderO, et al. The impact of sleep deprivation on hippocampal-mediated learning and memory in rats [J]. Bratisl Med J, 2017, 118(7): 408-416.
26 RechtschaffenA, BergmannB M. Sleep deprivation in the rat: an update of the 1989 paper [J]. Sleep, 2002, 25(1): 18-24.
27 SomarajanB I, KhandayM A, MallickB N. Rapid eye movement sleep deprivation induces neuronal apoptosis by noradrenaline acting on alpha1 adrenoceptor and by triggering mitochondrial intrinsic pathway [J]. Front Neurol, 2016, 7: 25.
28 HipólideD C, D'AlmeidaV, RaymondR, et al. Sleep deprivation does not affect indices of necrosis or apoptosis in rat brain [J]. Int J Neurosci, 2002, 112(2): 155-166.
29 SungY H. Study on emotion by rest time in mice with repetitive sleep deprivation [J]. J Exerc Rehabil, 2017, 13(2): 143-147.

[1]	李芳, 霍速, 杜巨豹, 刘秀贞, 李小爽, 宋为群. 经颅直流电刺激联合任务导向性康复训练对脊髓损伤大鼠前肢运动障碍的效果[J]. 《中国康复理论与实践》, 2023, 29(7): 777-781.
[2]	罗兰, 李璐, 金沐. 氙气后处理对脊髓缺血再灌注损伤的效果：Akt信号通路和自噬机制[J]. 《中国康复理论与实践》, 2023, 29(2): 174-181.
[3]	黄志霖, 徐发邵, 施静, 黄淦, 刘梅芳, 张霞辉. 线栓法建立卒中后吞咽障碍的大鼠模型[J]. 《中国康复理论与实践》, 2023, 29(10): 1147-1153.
[4]	秦彦强, 董浩, 孙迎春, 程先宽, 姚海江. 不同针刺方案对卒中后抑郁大鼠神经递质及相关炎性因子的影响[J]. 《中国康复理论与实践》, 2023, 29(1): 30-37.
[5]	缪培,张通,米海霞,张伟东. 不同线栓法复制局灶性脑缺血模型大鼠恢复期学习记忆能力的差异及其机制[J]. 《中国康复理论与实践》, 2022, 28(7): 789-796.
[6]	周小珏,冯婧,庞日朝,刘捷,张安仁. 隔日限食减轻脊髓损伤大鼠炎症反应的芳香烃受体/细胞因子信号传导抑制因子2/核转录因子-κB信号通路机制[J]. 《中国康复理论与实践》, 2022, 28(5): 544-551.
[7]	宋绍霏,侯园园,王云雷,张通. 异常光周期所致昼夜节律紊乱对大鼠腓肠肌时钟基因和葡萄糖摄取相关基因表达节律的影响[J]. 《中国康复理论与实践》, 2022, 28(5): 552-558.
[8]	黄菲菲,王万松,董晓阳,冯珍. 高压氧修复缺血再灌注损伤大鼠血脑屏障的自噬机制[J]. 《中国康复理论与实践》, 2022, 28(4): 415-420.
[9]	李童,方志鹏,邵玉萍,王平. 有氧运动对睡眠剥夺大鼠学习记忆及海马神经元突触可塑性的效果[J]. 《中国康复理论与实践》, 2022, 28(11): 1270-1277.
[10]	王静怡,尹杰,刘建成,庞日朝,王文春. 蜘蛛香环烯醚萜类成分对急性脊髓损伤大鼠神经细胞焦亡的影响[J]. 《中国康复理论与实践》, 2021, 27(6): 653-660.
[11]	周文美,陶陶,吴霜,王廷龙,杨正奕,张莹. 丰富环境对脑缺血再灌注损伤大鼠神经功能和缺血半暗带区葡萄糖代谢的影响[J]. 《中国康复理论与实践》, 2021, 27(5): 522-529.
[12]	王琼芬,王风波,王科,钟永强,王娇娇. 电针风池对急性脑梗死大鼠脑星形胶质细胞和神经元的保护效果[J]. 《中国康复理论与实践》, 2021, 27(3): 302-309.
[13]	李向哲,丁洁,王庆华,董传明,王彤,吴勤峰. 减重平板训练对脊髓损伤大鼠神经病理性疼痛及脊髓后角谷氨酸脱羧酶-65/67表达的影响[J]. 《中国康复理论与实践》, 2021, 27(2): 131-136.
[14]	艾坤,许明,刘琼,邓石峰,刘继生,祁芳,易细芹,瞿启睿,张泓. 电针对骶上脊髓损伤后逼尿肌亢进大鼠逼尿肌中环腺苷酸和蛋白激酶A含量及肌球蛋白轻链激酶磷酸化的影响[J]. 《中国康复理论与实践》, 2021, 27(2): 137-144.
[15]	凌梦钰,杨一卓,刘帅,叶超群. 运动训练对脑缺血再灌注大鼠认知功能及前额皮层神经元核抗原和突触素1表达的效果[J]. 《中国康复理论与实践》, 2021, 27(11): 1272-1281.

睡眠剥夺对大鼠血脑屏障的影响

Influence of Sleep Deprivation on Blood-brain Barrier in Rats

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0