远隔缺血处理对脑缺血再灌注大鼠认知功能障碍的神经保护作用

doi:10.3969/j.issn.1673-5765.2020.03.005

摘要/Abstract

摘要：

目的观察远隔缺血处理对缺血再灌注损伤所致认知障碍大鼠的神经保护作用及其作用机理。方法选用雄性SD大鼠，采用双侧颈动脉闭塞（bilateral common carotid arteries occlusion，BCCAO）方法建立脑缺血再灌注致血管性认知障碍（vascular cognitive impairment，VCI）动物模型，将大鼠随机分为假手术组、对照组（VCI组）及远隔缺血处理（remote ischemic conditioning，RIC）组（VCI+RIC组）。VCI 建模成功24 h后对RIC组大鼠进行RIC连续干预21 d，RIC期满后对三组大鼠采用Morris水迷宫实验进行定位航行实验和空间探索实验，定位航行实验连续进行5 d，记录第1天、第3天、第5天检测大鼠逃避潜伏期，第6天检测平台停留时间和穿越平台次数以评估认知功能。实验完成后在各组大鼠中随机选择5只取脑进行HE染色，电镜下观察各组大鼠脑白质及海马病理改变和神经元凋亡情况。结果 Morri s水迷宫实验结果显示第1天、第3天、第5天对照组及RI C组大鼠逃避潜伏期较假手术组均有延长（P＜0.05）。除假手术组外，另外两组大鼠逃避潜伏期时长随着训练次数增加逐渐缩短。第1天、第3天两组间差异无统计学意义，第5天RIC组成绩好于对照组（P＜0.05）。第6天空间探索实验中，目标象限停留时间假手术组与RIC组相近（47.2±10.2 s vs 41.2±9.7 s，P＞0.05），均优于对照组（33.5±11.3 s），差异均具有统计学意义；穿越平台次数假手术组与RIC组相近（5.3±1.6 vs 4.7±1.2， P＞0.05），均多于对照组（2.8±1.3），差异均具有统计学意义。在空间探索实验中，与假手术组比较，对照组大鼠的运动缺乏目的性而呈现出杂乱的曲线轨迹，RIC组相对于对照组具有明确的目的呈现出较规则的曲线。HE染色显示与假手术组（CA1：93.53±5.01；CA3：104.63±8.26）相比，对照组CA1区和CA3区存活的锥体神经元数目（CA1：51.03±4.95；CA3：78.53±5.31）明显减少（均P＜0.05）；而与对照组相比，RIC组大鼠CA1区和CA3区锥体神经元存活数目（CA1：80.57±7.30；CA3：92.43±8.16）明显增加（均P＜0.05）。结论脑缺血再灌注损伤可引起大鼠学习记忆障碍而导致VCI。RIC能够明显改善VCI，发挥神经保护作用。

文章导读： 远隔缺血处理可改善脑缺血再灌注损伤所致血管性认知障碍动物模型的认知障碍，发挥神经保护作用。

关键词: 远隔缺血处理; 脑缺血再灌注; 认知功能障碍; 神经保护

Abstract:

Objective To explore the neuroprotective effect of remote ischemic conditioning (RIC) and the corresponding mechanism in rats with vascular cognitive impairment (VCI) due to cerebral ischemia-reperfusion injury. Methods Male Sprague-Dawley rats were selected and randomly assigned to sham operation group (sham group), control group (VCI group) and RIC group (VCI+RIC group). The VCI animal models were established by bilateral common carotid arteries occlusion (BCCAO) method. The RIC was initiated after 24 hours of establishing the VCI models and lasted for 21 days only in RIC group. After the completion of RIC, the Morris water maze test was performed for place navigation test and space probe test in the three groups. In navigation test, the rats received 5 daily swim trials, the escape latency of rats were recorded on day 1, 3 and 5, and the platform stay time and the times of crossing platform of rats on day 6 were recorded to evaluate their cognitive function. After completing the whole procedure, 5 rats in each group were randomly selected to take brain tissue for HE staining. The pathological changes of brain white matter and hippocampus and neuron apoptosis were observed under the electron microscope. Results The Morris water maze test results showed that the escape latency of control group and RIC group were prolonged compared with sham group on day 1, 3, and 5 (P <0.05). Except sham group, the escape latency of the other two groups were gradually shortened with the increase of training times. There were no statistical difference between the two groups on day 1 and 3, and the performance of RIC group was better than that of control group on day 5 (P <0.05). In the space probe test on day 6, the sham group was similar to RIC group in the platform stay time (47.2±10.2 s vs 41.2±9.7 s, P >0.05), and they were better than control group (33.5±11.3 s) (both P <0.05). The sham group was similar to RIC group in the times of crossing platform (5.3±1.6 vs 4.7±1.2, P >0.05), and they were more than control group (2.8±1.3) (both P <0.05). In the probe test, compared with sham group, the control group showed a chaotic curve trajectory lacking purpose; compared with control group, the RIC group showed a more regular curve with a clear purpose. Compared with sham group (CA1: 93.53±5.01; CA3: 104.63±8.26), the number of surviving pyramidal neurons in CA1 and CA3 areas in control group (CA1: 51.03±4.95; CA3: 78.53±5.31) decreased significantly (both P <0.05); compared with control group, the survival number of surviving pyramidal neurons in CA1 and CA3 areas in RIC group (CA1: 80.57±7.30; CA3: 92.43±8.16) increased significantly (both P <0.05). Conclusions Cerebral ischemia-reperfusion injury can cause VCI presenting with learning and memory impairment. RIC can significantly improve VCI by playing neuroprotective effect.

Key words: Remote ischemic conditioning; Cerebral ischemia reperfusion; Cognitive impairment; Neuroprotective effect

徐耀铭，周文静，赵林，山花，赵文博，吉训明. 远隔缺血处理对脑缺血再灌注大鼠认知功能障碍的神经保护作用[J]. 中国卒中杂志, 2020, 15(03): 250-256.

XU Yao-Ming, ZHOU Wen-Jing, ZHAO Lin, SHAN Hua, ZHAO Wen-Bo,JI Xun-Ming. Neuroprotective Effect of Remote Ischemic Conditioning on Cognitive Impairment due to Cerebral Ischemia-reperfusion Injury in Rats[J]. Chinese Journal of Stroke, 2020, 15(03): 250-256.

参考文献

[1] HACHINSKI V，IADECOLA C，PETERSEN R C，

et al. National Institute of Neurological Disorders

and Stroke-Canadian Stroke Network vascular

cognitive impairment harmonization standards[J].

Stroke，2006，37（9）：2220-2241.

[2] DAVE K R，SAUL I，PRADO R，et al. Remote

organ ischemic preconditioning protect brain from

ischemic damage following asphyxial cardiac

arrest[J]. Neurosci Lett，2006，404（1-2）：170-175.

[3] JIWA N S，GARRARD P，HAINSWORTH A

H. Experimental models of vascular dementia

and vascular cognitive impairment：a systematic

review[J]. J Neurol Sci，2010，299（1-2）：188-192.

[4] REN C，GAO X，STEINBERG G K，et al. Limb

remote-preconditioning protects against focal

ischemia in rats and contradicts the dogma of

therapeutic time windows for preconditioning[J].

Neuroscience，2008，151（4）：1099-1103.

[5] KUMAR R，BUKOWSKI M J，WIDER J M，et al.

Mitochondrial dynamics following global cerebral

ischemia[J]. Mol Cell Neurosci，2016，76：68-75.

[6] FAN M，SONG C，WANG T，et al. Protective effects

of lithium chloride treatment on repeated cerebral

ischemia-reperfusion injury in mice[J]. NeurolSci，

2015，36（2）：315-321.

[7] MINGYUE F，WEI J，HAIFENG Z，et al. Lithium

chloride administration prevents spatial learning and

memory impairment in repeated cerebral ischemiareperfusion

mice by depressing apoptosis and

increasing BDNF expression in hippocampus[J].

Behav Brain Res，2015，15：399-406.

[8] VORHEES C V，WILLIAMS M T. Morris water

maze：procedures for assessing spatial and related

forms of learning and memory[J]. Nat Protoc，2006，

1（2）：848-858.

[9] D'HOOGE R，de DEYN P P. Applications of the

Morris water maze in the study of learning and

memory[J]. Brain Res Brain Res Rev，2001，36（1）：

60-90.

[10] AL-AMIN M M，RAHMAN M M，KHAN F R，et

al. Astaxanthin improves behavioral disorder and

oxidative stress in prenatal valproic acid-induced

mice model of autism[J]. Behav Brain Res，2015，

286：112-121.

[11] TAPURIA N，KUMAR Y，HABIB M M，et al.

Remote ischemic preconditioning a novel protective

method from ischemic reperfusion injury-a review[J].

Surg Res，2008，150（2）：304-330.

[12] LIM S Y，YELLON D M，HAUSENLOY D J.

The neural and humoral pathways in remote limb

ischemic preconditioning[J]. Basic Res Cardiol，2010，

105（5）：651-655.

[13] MEDVEDEVA Y V，JI S G，YIN H Z，et al.

Differential vulnerability of CA1 versus CA3

pyramidal neurons after ischemia：possible

relationship to sources of Zn2+ accumulation and its

entry into and prolonged effects on mitochondria[J].

J Neurosci，2017，37（3）：726-737.

[14] YIN B，BARRIONUEVO G，BATINICHABERLE

I，et al. Differences in reperfusioninduced

mitochondrial oxidative stress and cell

death between hippocampal CA1 and CA3 subfields

are due to the mitochondrial thioredoxin system[J].

Antioxid Redox Signal，2017，27（9）：534-549.

[15] SUN L N，SHEN J，SU F，et al. Bicyclol attenuates

oxidative stress and neuronal damage following

transient forebrain ischemia in mouse cortex and

hippocampus[J]. Neurosci Lett，2009，459（2）：84-

87.

[16] WANG J Y，SHEN J，GAO Q，et al. Ischemic

postconditioning protects against global cerebral

ischemia/reperfusion-induced injury in rats[J].

Stroke，2008，39（3）：983-990.

[17] LI P，SU L，LI X，et al. Remote limb ischemic

postconditioning protects mouse brain against cerebral ischemia/reperfusion injury via

upregulating expression of Nrf2，HO-1，NQO-1 in

mice [J]. Int J Neurosci，2016，126（6）：552-559.

[18] PENG B，GUO Q L，HE Z J，et al. Remote

ischemic postconditioning protects the brain from

global cerebral ischemia/reperfusion injury by upregulating

endothelial nitric oxide synthase through

the PI3K/Akt pathway[J]. Brain Res，2012，1445：

92-102.