电损毁外侧缰核对帕金森病模型大鼠空间学习记忆功能的影响及其机制

doi:10.13481/j.1671-587X.20210505

摘要/Abstract

摘要： 目的

观察电损毁外侧缰核（LHb）对帕金森病（PD）模型大鼠空间学习记忆功能的影响，阐明LHb在PD大鼠认知功能障碍中的作用及其相关机制。

方法

30只成年雄性SD大鼠随机分为对照组、PD组和PD+LHb损毁组，每组10只，除对照组外，其余2组大鼠制备PD模型，PD+LHb损毁组PD大鼠造模2周后电损毁LHb，造模后第4周进行相关检测；旷场实验观察3组大鼠穿行格数和站立次数；Morris水迷宫实验观察3组大鼠寻找水下平台游泳距离和撤除平台后在目标象限内游泳距离百分率；神经化学法观察3组大鼠脑内内侧前额叶皮层（mPFC）和海马组织中5-羟色胺（5-HT）水平；免疫组织化学和组织学染色法检测脑组织中多巴胺（DA）能神经元的缺失程度和LHb电损毁情况。

结果

旷场实验，与对照组比较，PD组大鼠的穿行格数和站立次数明显减少（P<0.01）；与PD组比较，PD+LHb损毁组大鼠的穿行格数和站立次数差异无统计学意义（P>0.05）。Morris水迷宫实验，与对照组比较，训练第2、3、4和5天PD组大鼠在定位航行实验中寻找水下平台游泳距离明显增加（P<0.01），撤除平台后空间探索实验中目标象限内游泳距离百分率明显降低（P<0.05），但对照组和PD组大鼠脑内mPFC和海马组织中5-HT水平差异无统计学意义（P>0.05）；与PD组比较，训练第2、3、4和5天PD+LHb损毁组大鼠寻找水下平台游泳距离增加（P<0.01），且撤除平台后目标象限内游泳距离百分率也进一步降低（P<0.05），脑内mPFC和海马组织中5-HT水平明显降低（P<0.01）。免疫组织化学检测，PD大鼠损毁侧黑质致密部DA能神经元完全缺失，且腹侧被盖区DA能神经元较未损毁侧明显减少（P<0.01）；组织学确认LHb被部分损毁。

结论

损毁LHb可加重PD大鼠空间学习记忆功能损伤，其机制与改变相关脑区中5-HT递质水平有关。

关键词: 外侧缰核, 空间学习记忆, 帕金森病, Morris水迷宫

Abstract: Objective

To observe the effect of electrical lesions of the lateral habenular nucleus （LHb） on the spatial learning and memory functions in the rats with Parkinson’s disease （PD）， and to clarify the role of LHb in the cognitive dysfunction in the PD rats and related mechanism.

Methods

Thirty adult male SD rats were randomly divided into control group， PD group and PD+LHb lesion group，and there were 10 rats in each group. Except control group， the rats in other two groups were used to establish the PD models. The PD rats in PD+LHb lesion group were lesioned using direct current stimulation two weeks after modeling，and the related tests were performed at the 4th week after modeling. The numbers of crossed squares and rearings of the rats in three groups were observed by open field test. The distances to find the hidden platform and the percentages of swimming distance in target quadrant of the rats in three groups were observed by Morris water maze test. The levels of serotonin （5-HT） in the medial prefrontal cortex （mPFC） and hippocampus tissue of the rats in three groups were assessed by neurochemical method. The degeneration of dopamine （DA） neurons in the brain tissue and LHb electrical lesioning were confirmed by the immunohistochemical and histological staining methods.

Results

The results of open field test showed that compared with control group， the numbers of crossed squares and rearings of the rats in PD group were significantly decreased （P<0.01），and the number of crossed squares and rearings of the rts in PD+LHb lesion group had no significantly differences（P>0.05）.The results of Morris water maze test showed that compared with control group， the distances to find the hidden platform in navigation trial of the rats in PD group on the 2nd， 3rd，4th and 5th days were significantly increased （P<0.01）， and the percentages of swimming distance in target quadrant after withdrawal of platform in space exploration experiment were significantly decreased （P<0.05）； but there were no significant differences in the 5-HT levels in the mPFC and hippocampus tissues of the rats between two groups（P>0.05）. There were no significant differences in the numbers of crossed squares and rearings of the rats between PD and PD+LHb lesion groups； compared with PD group， the distances to find the hidden platform of the rats in PD+LHb group on the 2nd， 3rd，4th and 5th days were significantly increased （P<0.01），the percentages of swimming distance in target quadrant after withdrawal of platform were significantly decreased （P<0.05）， and the 5-HT levels in mPFC and hippocampus tissues were significantly decreased （P<0.01）. The immunohistochemical staining results showed the DA neurons of the lesioned side of the PD model rats in the substantia nigra pars compacta were missing， and the number of DA neurons in ventral tegmental area was lower than that in unlesioned side （P<0.01）.The histological staining results showed the LHb was partially lesioned.

Conclusion

LHb lesions can aggravate the impairment of spatial learning and memory functions of the rats with PD， and its mechanism may be related to the changes of levels of 5-HT transmitter in the related brain regions.

Key words: lateral habenular nucleus, spatial learning and memory, Parkinson’s disease, Morris water maze

中图分类号:

R338.8

韩玲娜,王春雷,常永丽,原丽,刘小静. 电损毁外侧缰核对帕金森病模型大鼠空间学习记忆功能的影响及其机制[J]. 吉林大学学报(医学版), 2021, 47(5): 1108-1115.

Lingna HAN,Chunlei WANG,Yongli CHANG,Li YUAN,Xiaojing LIU. Effect of electrical lesions of lateral habenular nucleus on spatial learning and memory functions in rats with Parkinson’s disease and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2021, 47(5): 1108-1115.

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Group	Number of crossed squares	Number of rearings
Control	74.70±9.39	18.90±2.54
PD	37.00±5.42*	9.20±1.15*
PD+LHb lesion	34.00±6.27	7.90±1.15

Group	Distance to find thidden platform (l/cm)					Percentage of swimming distance in target quadrant （η/%）
Group	(t/d) 1	2	3	4	5	Percentage of swimming distance in target quadrant （η/%）
Control	1 770.34±104.99	784.44±54.70	497.88±36.59	268.04±34.08	189.70±26.83	57.70±4.40
PD	1 915.47±121.33	1 159.97±52.89^**	784.37±54.37^**	513.95±33.70^**	373.79±40.18^**	42.64±4.18^*
PD+LHb lesion	1 843.32±77.28	1 670 .01±87.40^△△	1 123.77±99.67^△△	956.47±66.03^△△	643.79±49.17^△△	26.65±3.97^△

Group	Level of 5-HT
Group	mPFC	Hippocampus
Control	641.44±73.87	500.45±41.80
PD	492.50±25.27	450.57±49.89
PD+LHb lesion	269.55±34.47*	214.24±27.55*

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