电针预处理对脑缺血再灌注大鼠行为学及外周血内皮祖细胞的影响

doi:10.3877/cma.j.issn.2095-9141.2024.02.002

中华神经创伤外科电子杂志 ›› 2024, Vol. 10 ›› Issue (02) : 71 -77. doi: 10.3877/cma.j.issn.2095-9141.2024.02.002

基础研究

电针预处理对脑缺血再灌注大鼠行为学及外周血内皮祖细胞的影响

张艺¹, 任秀君¹, 郭孟玮¹, 赵雅芳¹, 李一凡¹, 李佳阳¹, 任晓暄¹, 邬继红¹, 卢海洋²^,^†(

)

^1. 102488　北京，北京中医药大学针灸推拿学院
^2. 100050　北京，首都医科大学附属北京友谊医院麻醉科

收稿日期:2023-12-26 出版日期:2024-04-15
通信作者: 卢海洋

Effects of electroacupuncture preconditioning on behavioural and peripheral blood EPCs in cerebral ischaemia-reperfusion injury rats

Yi Zhang¹, Xiujun Ren¹, Mengwei Guo¹, Yafang Zhao¹, Yifan Li¹, Jiayang Li¹, Xiaoxuan Ren¹, Jihong Wu¹, Haiyang Lu²^,^†()

^1. School of Acupuncture-Moxibustion and Tuina, Beijing University of Traditional Chinese Medicine, Beijing 102488, China
^2. Department of Anesthesiology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China

Received:2023-12-26 Published:2024-04-15
Corresponding author: Haiyang Lu
Supported by:
National Natural Science Foundation of China(82271307); Beijing Natural Science Foundation(7212167)

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引用本文：

张艺, 任秀君, 郭孟玮, 赵雅芳, 李一凡, 李佳阳, 任晓暄, 邬继红, 卢海洋. 电针预处理对脑缺血再灌注大鼠行为学及外周血内皮祖细胞的影响[J]. 中华神经创伤外科电子杂志, 2024, 10(02): 71-77.

Yi Zhang, Xiujun Ren, Mengwei Guo, Yafang Zhao, Yifan Li, Jiayang Li, Xiaoxuan Ren, Jihong Wu, Haiyang Lu. Effects of electroacupuncture preconditioning on behavioural and peripheral blood EPCs in cerebral ischaemia-reperfusion injury rats[J]. Chinese Journal of Neurotraumatic Surgery(Electronic Edition), 2024, 10(02): 71-77.

目的

观察电针预处理对脑缺血再灌注损伤（CIRI）大鼠行为学及外周血内皮祖细胞（EPCs）水平的影响。

方法

45只成年雄性SD大鼠随机分为假手术组、模型组和电针预处理组（EA组），每组15只。模型组于造模前将大鼠束缚14 d，采用Longa线栓法制备大脑中动脉缺血再灌注模型；假手术组除不插入线栓外，余同模型组；EA组在造模前14 d行电针预处理，余同模型组。造模当天监测大鼠脑血流量，造模后1 d计算脑梗死体积，造模后1、3、7、21、28 d分别采用改良神经功能缺损评分（mNSS）、旷场实验和Morris水迷宫实验对大鼠相关指标进行检测，取外周血采用流式细胞术测定EPCs水平。

结果

模型组和EA组造模过程中各时间点的脑血流量明显低于假手术组，EA组在线栓即刻及再灌注30 min的脑血流量高于模型组，差异均有统计学意义（P<0.05）。模型组和EA组脑梗死体积百分比显著高于假手术组，EA组脑梗死体积百分比显著低于模型组，差异均有统计学意义（P<0.05）。EA组造模后1、3、7及21 d的mNSS评分均低于模型组，差异有统计学意义（P<0.05）。EA组大鼠旷场实验的平均速度在造模后1 d明显高于模型组，目标象限滞留时间百分比在造模后3 d高于模型组，差异有统计学意义（P<0.05）。外周血EPCs水平在造模后2 d高于模型组，差异均有统计学意义（P<0.05）。

结论

电针预处理一定程度上可以改善急性期CIRI大鼠的脑血流状态，减轻神经功能损伤，提升学习记忆能力和运动能力，提高外周血中EPCs水平。

关键词: 电针预处理, 脑缺血再灌注损伤, 大脑中动脉栓塞, 内皮祖细胞

Objective

To observe the effects of electroacupuncture pretreatment on behavioral and peripheral blood endothelial progenitor cells (EPCs) levels in rats with cerebral ischemia-reperfusion injury (CIRI).

Methods

Forty-five adult male SD rats were randomly divided into the sham group, the model group and the electroacupuncture pretreatment group (EA group), with 15 rats in each group. Rats in the model group were restrained for 14 d before surgery, and the middle cerebral artery occlusion ischemia-reperfusion model was established by Longa thread-occluded method; rats in the sham group were treated as the same as the rats in the model group except for not inserting thread plug; rats in the EA group were pretreated with EA 14 d before surgery, others were the same as those in the model group. Cerebral blood flow was monitored on the day of modelling, and infarct volume was calculated 1 d after modelling. At different time points after modelling, the modified neurological severity score (mNSS), the open-field experiment and the Morris water maze experiment were used to detect the relevant indexes in rats, and peripheral blood was taken to determine the levels of EPCs by flow cytometry.

Results

The cerebral blood flow at each time point during the modeling process in the model group and EA group was significantly lower than that in the sham surgery group. The cerebral blood flow in the EA group was higher than that in the model group immediately after thrombus insertion and 30 min of reperfusion, and the differences were statistically significant (P<0.05). The cerebral infarction volume percentage in the model group and EA group was significantly higher than that in the sham surgery group, while the cerebral infarction volume percentage in the EA group was significantly lower than that in the model group, with statistical significance (P<0.05). The mNSS scores of the EA group at 1, 3, 7, and 21 d after modeling were lower than those of the model group, and the difference was statistically significant (P<0.05). The average speed of the open field experiment in EA group rats was significantly higher than that of the model group 1 d after modeling, and the percentage of target quadrant retention time was higher than that of the model group 3 d after modeling, the differences were statistically significant (P<0.05). The peripheral blood EPCs level was higher than that of the model group 2 d after modeling, and the differences were statistically significant (P<0.05).

Conclusion

Electroacupuncture pretreatment can improve the cerebral blood flow status of acute phase CIRI rats to a certain extent, alleviate neurological damage, enhance learning and memory abilities and motor abilities, and increase the level of EPCs in peripheral blood.

Key words: Electroacupuncture pretreatment, Cerebral ischemia-reperfusion injury, Middle cerebral artery embolism, Endothelial progenitor cells

图1 3组大鼠造模后的生存曲线

Fig.1 Survival curves of three groups of rats after modeling

表1 3组大鼠脑血流量比较（%，

±s）

Tab.1 Comparison of cerebral blood flow after modeling in three groups of rats (%, Mean±SD)

组别	只数	线栓即刻	缺血后2 h	再灌注10 min	再灌注20 min	再灌注30 min	F值	P值
假手术组	6	97.11±6.56	91.67±12.12	93.87±11.47	95.94±11.60	95.43±9.56	1.976	0.162
模型组	6	44.62±5.44^a	45.75±3.33^a	59.92±4.80^a	60.33±4.83^a	57.00±3.87^a	50.724	<0.001
EA组	6	52.13±3.88^ab	47.41±3.33^a	66.48±2.54^a	65.74±1.75^a	67.83±4.02^ab	106.045	<0.001
F值		137.810	60.161	30.194	34.293	48.095
P值		<0.001	<0.001	<0.001	<0.001	<0.001

表1 3组大鼠脑血流量比较（%，

±s）

Tab.1 Comparison of cerebral blood flow after modeling in three groups of rats (%, Mean±SD)

组别	只数	线栓即刻	缺血后2 h	再灌注10 min	再灌注20 min	再灌注30 min	F值	P值
假手术组	6	97.11±6.56	91.67±12.12	93.87±11.47	95.94±11.60	95.43±9.56	1.976	0.162
模型组	6	44.62±5.44^a	45.75±3.33^a	59.92±4.80^a	60.33±4.83^a	57.00±3.87^a	50.724	<0.001
EA组	6	52.13±3.88^ab	47.41±3.33^a	66.48±2.54^a	65.74±1.75^a	67.83±4.02^ab	106.045	<0.001
F值		137.810	60.161	30.194	34.293	48.095
P值		<0.001	<0.001	<0.001	<0.001	<0.001

表2 3组大鼠造模后1 d的脑梗死体积百分比比较（%，

±s）

Tab.2 Comparison of the percentage of cerebral infarction volume in three groups of rats after 1 d of modeling (%, Mean±SD)

组别	只数	梗死体积
假手术组	9	0.00±0.00
模型组	9	29.01±3.52^a
EA组	9	23.67±3.53^ab
F值		232.017
P值		0.001

表2 3组大鼠造模后1 d的脑梗死体积百分比比较（%，

±s）

Tab.2 Comparison of the percentage of cerebral infarction volume in three groups of rats after 1 d of modeling (%, Mean±SD)

组别	只数	梗死体积
假手术组	9	0.00±0.00
模型组	9	29.01±3.52^a
EA组	9	23.67±3.53^ab
F值		232.017
P值		0.001

表3 3组大鼠造模后各时间点的mNSS评分比较（分，

±s）

Tab.3 Comparison of mNSS scores at different time points after modeling in three groups of rats (score, Mean±SD)

组别	只数	造模后1 d	造模后3 d	造模后7 d	造模后21 d	造模后28 d	F值	P值
假手术组	6	0	0	0	0	0	-	-
模型组	6	12.00±1.41^a	9.00±0.58^a	7.00±1.15^a	3.83±1.46^a	2.33±1.60^a	54.772	<0.001
EA组	6	9.33±0.94^ab	7.50±0.76^ab	4.83±1.57^ab	2.33±1.25^ab	1.33±0.94	50.638	<0.001
F值		206.154	380.455	50.620	15.150	5.968
P值		<0.001	<0.001	<0.001	<0.001	0.012

表3 3组大鼠造模后各时间点的mNSS评分比较（分，

±s）

Tab.3 Comparison of mNSS scores at different time points after modeling in three groups of rats (score, Mean±SD)

组别	只数	造模后1 d	造模后3 d	造模后7 d	造模后21 d	造模后28 d	F值	P值
假手术组	6	0	0	0	0	0	-	-
模型组	6	12.00±1.41^a	9.00±0.58^a	7.00±1.15^a	3.83±1.46^a	2.33±1.60^a	54.772	<0.001
EA组	6	9.33±0.94^ab	7.50±0.76^ab	4.83±1.57^ab	2.33±1.25^ab	1.33±0.94	50.638	<0.001
F值		206.154	380.455	50.620	15.150	5.968
P值		<0.001	<0.001	<0.001	<0.001	0.012

表4 3组大鼠旷场实验平均速度比较（cm/s，

±s）

Tab.4 Comparison of average speed in open field experiments of three groups of rats (cm/s, Mean±SD)

组别	只数	造模后1 d	造模后3 d	造模后7 d	造模后21 d	造模后28 d	F值	P值
假手术组	6	5.60±0.45	5.65±2.17	5.64±1.03	5.93±0.80	6.12±1.98	0.087	0.985
模型组	6	2.58±0.76^a	3.17±1.00	3.95±1.00^a	5.41±2.63	5.43±2.52	8.571	0.002
EA组	6	3.84±1.05^ab	4.10±1.36	4.65±0.80	5.62±2.77	5.64±1.33	3.225	0.051
F值		18.311	3.109	3.982	0.040	0.154
P值		<0.001	0.074	0.041	0.961	0.858

表4 3组大鼠旷场实验平均速度比较（cm/s，

±s）

Tab.4 Comparison of average speed in open field experiments of three groups of rats (cm/s, Mean±SD)

组别	只数	造模后1 d	造模后3 d	造模后7 d	造模后21 d	造模后28 d	F值	P值
假手术组	6	5.60±0.45	5.65±2.17	5.64±1.03	5.93±0.80	6.12±1.98	0.087	0.985
模型组	6	2.58±0.76^a	3.17±1.00	3.95±1.00^a	5.41±2.63	5.43±2.52	8.571	0.002
EA组	6	3.84±1.05^ab	4.10±1.36	4.65±0.80	5.62±2.77	5.64±1.33	3.225	0.051
F值		18.311	3.109	3.982	0.040	0.154
P值		<0.001	0.074	0.041	0.961	0.858

表5 3组大鼠目标象限滞留时间百分比比较（%，

±s）

Tab.5 Comparison of the percentage of target quadrant retention time among three groups of rats (%, Mean±SD)

组别	只数	造模后3 d	造模后7 d	造模后21 d	造模后28 d	F值	P值
假手术组	6	35.25±10.17	21.87±2.87	18.83±2.70	18.05±5.63	12.000	<0.001
模型组	6	19.93±3.95^a	19.31±2.36	18.93±6.48	17.28±8.31	0.510	0.683
EA组	6	29.43±4.65^b	20.67±3.59	17.68±4.86	17.40±4.97	5.608	0.011
F值		6.347	0.913	0.099	0.020
P值		0.010	0.422	0.906	0.980

表5 3组大鼠目标象限滞留时间百分比比较（%，

±s）

Tab.5 Comparison of the percentage of target quadrant retention time among three groups of rats (%, Mean±SD)

组别	只数	造模后3 d	造模后7 d	造模后21 d	造模后28 d	F值	P值
假手术组	6	35.25±10.17	21.87±2.87	18.83±2.70	18.05±5.63	12.000	<0.001
模型组	6	19.93±3.95^a	19.31±2.36	18.93±6.48	17.28±8.31	0.510	0.683
EA组	6	29.43±4.65^b	20.67±3.59	17.68±4.86	17.40±4.97	5.608	0.011
F值		6.347	0.913	0.099	0.020
P值		0.010	0.422	0.906	0.980

表6 3组大鼠外周血EPCs水平比较（%，

±s）

Tab.6 Comparison of peripheral blood EPCs levels in three groups of rats (%, Mean±SD)

组别	只数	造模后2 d	造模后6 d	造模后20 d	造模后27 d	F值	P值
假手术组	6	9.47±0.74	9.37±1.99	10.07±1.99	10.67±1.31	0.430	0.735
模型组	6	4.73±1.79^a	5.55±1.71^a	8.07±0.76	9.23±2.69	6.436	0.007
EA组	6	6.85±0.71^ab	7.80±0.70	8.93±1.44	9.43±3.29	1.994	0.165
F值		14.149	5.529	2.632	0.357
P值		<0.001	0.016	0.105	0.705

表6 3组大鼠外周血EPCs水平比较（%，

±s）

Tab.6 Comparison of peripheral blood EPCs levels in three groups of rats (%, Mean±SD)

组别	只数	造模后2 d	造模后6 d	造模后20 d	造模后27 d	F值	P值
假手术组	6	9.47±0.74	9.37±1.99	10.07±1.99	10.67±1.31	0.430	0.735
模型组	6	4.73±1.79^a	5.55±1.71^a	8.07±0.76	9.23±2.69	6.436	0.007
EA组	6	6.85±0.71^ab	7.80±0.70	8.93±1.44	9.43±3.29	1.994	0.165
F值		14.149	5.529	2.632	0.357
P值		<0.001	0.016	0.105	0.705

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Effects of electroacupuncture preconditioning on behavioural and peripheral blood EPCs in cerebral ischaemia-reperfusion injury rats

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Effects of electroacupuncture preconditioning on behavioural and peripheral blood EPCs in cerebral ischaemia-reperfusion injury rats