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

doi:10.3969/j.issn.1006-9771.2022.07.008

摘要/Abstract

摘要：

目的探讨线栓法永久性脑缺血和缺血再灌注大鼠模型在3周自然恢复状态下学习记忆功能的差异及其脑结构变化。

方法 SPF级雄性Sprague-Dawley大鼠21只随机分为假手术组(n = 7)、永久性缺血组(n = 7)和缺血再灌注组(n = 7)。后两组线栓法制备左侧大脑中动脉闭塞(MCAO)模型,缺血再灌注组于缺血90 min恢复血供。术后16~22 d行Morris水迷宫实验,记录定位航行实验逃避潜伏期,空间探索实验中第一次到达平台所在位置的潜伏期、平台象限游泳时间比及距离比、边界游泳时间比及距离比、游泳路径。术后22 d各组取Longa评分相近的4只大鼠磁共振弥散张量成像(DTI)测量缺血皮质、纹状体、海马及其对侧相应区域的各向异性分数(FA)和梗死体积,计算FA比(rFA)和梗死体积比。

结果定位航行实验中,三组潜伏期均随训练时间增加而下降(F = 36.202, P < 0.001),但永久性缺血组各时间点间无显著性差异(F = 4.004, P > 0.05);永久性缺血组各时间点潜伏期均大于另两组(P < 0.05)。空间探索实验中,两模型组边界游泳时间比和距离比均显著大于假手术组(P < 0.017),永久性缺血组大鼠潜伏期大于假手术组(P < 0.017);缺血再灌注组和假手术组的游泳路径更优。两模型组左侧皮质、纹状体区FA、rFA均较假手术组下降(P < 0.05);永久性缺血组左侧皮质FA、rFA及左侧纹状体FA均低于缺血再灌注组(P < 0.05)。永久性缺血组梗死体积和梗死体积比明显大于缺血再灌注组(t > 4.185, P < 0.01)。皮质、纹状体左侧FA、rFA与逃避潜伏期、边界时间比和距离比相关(|r| > 0.498, P < 0.05);脑梗死体积和百分比与逃避潜伏期、边界时间比相关(|r| > 0.538, P < 0.05)。

结论永久性脑缺血大鼠模型认知功能障碍更严重,与梗死体积更大,皮质、纹状体神经纤维结构破坏更严重有关。

关键词: 脑缺血, 永久性缺血, 缺血再灌注, 学习记忆, 大鼠

Abstract:

Objective To explore the difference of learning and memory function between permanent cerebral ischemia and ischemia-reperfusion rat models by filament-occluded method after three weeks of natural recovery, and to observe the changes of brain structure.

Methods A total of 21 SPF male Sprague-Dawley rats were randomly divided into sham group (n = 7), permanent ischemia group (n = 7) and ischemia-reperfusion group (n = 7). The latter two group underwent left middle cerebral artery occlusion, and the ischemia-reperfusion group was reperfused after 90 minutes of ischemia. All the rats were tested with Morris Water Maze 16 to 22 days after modeling, to record escape latency in the navigation experiment, as well as first latency, platform quadrant swimming time ratio and distance ratio, boundary swimming time ratio and distance ratio, average speed, and the swimming path in the space exploration experiment. On the 22nd day after modeling, four rats with similar Longa scores in each group were scanned by magnetic resonance by diffusion tensor imaging, to measure the fractional anisotropy (FA) of cerebral ischemic cortex, striatum, hippocampus and their corresponding contralateral regions, to calculate ratio of FA (rFA); as well as the volume of infarct (VI) and ratio of VI (rVI).

Results For navigation experiment, the escape latency decreased in all the groups with the time (F = 36.202, P < 0.001), but it was not significant for the permanent ischemia group (F = 4.004, P > 0.05). The escape latency was longer in the permanent ischemia group than in the other two groups at each time points (P < 0.05). For space exploration experiment, the boundary time ratio and distance ratio were more in the permanent ischemia group and the ischemia-reperfusion group than in the sham group (P < 0.017), and the first latency was longer in the permanent ischemia group than in the sham group (P < 0.017). The swimming path of ischemia-reperfusion group and sham operation group was better than that of the permanent ischemia group. FA and rFA in left cortex and striatum were less in the permanent ischemia group and the ischemia-reperfusion group than in the sham group (P < 0.05), while FA and rFA of left cortex and FA of left striatum were less in the permanent ischemia group than in the ischemia-reperfusion group (P < 0.05). The VI and rVI were larger in the permanent ischemia group than in the ischemia-reperfusion group (t > 4.185, P < 0.01). FA and rFA of left cortex and striatum correlated with escape latency, boundary time ratio and distance ratio (|r| > 0.498, P < 0.05). The VI and rVI correlated with escape latency and boundary time ratio (|r| > 0.538, P < 0.05).

Conclusion Cognitive impairment is more severe in the rat model of permanent cerebral ischemia, which may relate to the larger damage of structure of nerve fibers in ischemic cortex and striatum, and larger infarct size.

Key words: cerebral ischemia, permanent ischemia, ischemia-reperfusion, learning and memory, rats

中图分类号:

R743.3

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.

MIAO Pei,ZHANG Tong,MI Haixia,ZHANG Weidong. Learning and memory ability and its mechanism in rats with focal cerebral ischemia induced by two filament-occluded methods[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(7): 789-796.

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组别	n	第1天	第2天	第3天	第4天	第5天	F值	P值
假手术组	7	48.457±6.43	19.214±12.08^a	12.117±4.00^a	11.099±3.92^a	6.900±1.64^a,b,c,d	44.268	< 0.001
缺血再灌注组	7	62.264±18.56	31.089±24.55^a	26.210±26.22^a	23.166±23.87^a,b	18.526±22.75^a,b	20.376	< 0.001
永久性缺血组	7	81.647±13.73^e,f	72.447±24.47^e,f	64.357±24.90^a,e,f	58.917±26.99^e,f	51.183±37.72^a,e,f	4.004	0.058
F值		10.165	12.172	11.590	9.889	5.696
P值		0.001	< 0.001	0.001	0.001	0.012

组别	n	潜伏期/s	象限时间比/%	象限距离比/%	边界时间比/%	边界距离比/%	速度/m·s^-1
假手术组	7	6.866±3.80	33.921±7.48	28.747±4.69	7.909±3.40	5.211±2.36	0.753±0.16
缺血再灌注组	7	17.101±9.21	30.844±9.76	33.734±9.92	20.759±11.51^a	29.241±30.19^a	0.783±0.13
永久性缺血组	7	55.349±42.68^a	26.346±5.55	27.753±5.33	46.163±35.48^a	42.847±37.79^a	0.810±0.11
χ²值		12.475	4.148	2.167	12.632	11.688	0.925
P值		0.002	0.126	0.338	0.002	0.003	0.630

组别	n	左侧FA	右侧FA	rFA
假手术组	4	0.212±0.01	0.228±0.03	0.937±0.07
缺血再灌注组	4	0.095±0.01^a	0.229±0.02	0.421±0.06^a
永久性缺血组	4	0.069±0.01^a,b	0.216±0.04	0.321±0.04^a,b
F值		251.107	0.237	133.492
P值		< 0.001	0.794	< 0.001

组别	n	左侧FA	右侧FA	rFA
假手术组	4	0.242±0.02	0.262±0.03	0.930±0.07
缺血再灌注组	4	0.121±0.01^a	0.311±0.04	0.396±0.08^a
永久性缺血组	4	0.091±0.01^a,b	0.276±0.03	0.328±0.02^a
F值		96.358	2.235	114.896
P值		< 0.001	0.163	< 0.001

组别	n	左侧FA	右侧FA	rFA
假手术组	4	0.223±0.08	0.225±0.07	1.000±0.08
缺血再灌注组	4	0.235±0.04	0.251±0.03	0.942±0.17
永久性缺血组	4	0.241±0.03	0.236±0.06	1.061±0.25
F值		0.133	0.214	0.445
P值		0.877	0.811	0.654