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

doi:10.3969/j.issn.1006-9771.2021.03.009

摘要/Abstract

摘要： 目的

探讨电针风池穴对急性脑梗死大鼠脑星形胶质细胞和神经元的调节作用。

方法

清洁级Sprague-Dawley雄性大鼠64只，随机数字表法分为假手术组(n = 16)、模型组(n = 16)、非穴组(n = 16)和风池组(n = 16)。后三组采用改良线栓法制备急性脑梗死模型，后两组于造模后分别电针风池旁非穴部位和风池，共7 d。治疗后进行神经功能缺损评分；取脑组织测量脑梗死面积和脑组织含水量；HE染色观察海马组织病理学改变；ELISA法检测海马组织中脑源性神经营养因子(BDNF)、胶质纤维酸性蛋白(GFAP)、神经元特异性烯醇化酶(NSE)的表达水平；采用TUNEL法检测海马神经元细胞凋亡数；采用Western blotting和逆转录实时定量聚合酶链反应(RT-qPCR)检测脑匀浆caspase-3和Bcl-2水平。

结果

与假手术组相比，其他三组神经功能缺损评分、脑梗死面积和含水量升高(P < 0.05)，海马组织中GFAP和NSE水平升高(P < 0.05)，BDNF降低(P < 0.05)，海马神经元凋亡细胞数升高(P < 0.05)，caspase-3表达升高(P < 0.05)，Bcl-2表达降低(P < 0.05)；与模型组和非穴组相比，风池组神经行为学评分、脑梗死面积和含水量降低(P < 0.05)，GFAP和NSE水平降低(P < 0.05)，BDNF升高(P < 0.05)，神经元凋亡细胞数减少(P < 0.05)，caspase-3表达降低(P < 0.05)，Bcl-2表达增加(P < 0.05)。

结论

电针风池穴可改善急性脑梗死大鼠脑组织损伤，调节脑组织GFAP、NSE和BDNF表达水平，降低神经元凋亡水平。

关键词: 脑梗死, 电针, 星形胶质细胞, 神经元, 大鼠

Abstract: Objective

To investigate the effect of electroacupuncture at Fengchi (GB20) on astrocytes and neurons in rats with acute cerebral infarction.

Methods

A total of 64 male Sprague-Dawley rats were randomly divided into sham group (n = 16), model group (n = 16), non-acupoint group (n = 16) and Fengchi group (n = 16). The latter three groups were modeled with middle cerebral artery occlusion (MCAO), and the latter two groups were treated with electroacupuncture after modeling, at the point near Fengchi and Fengchi, respectively, for seven days. All the rats were assessed with neurological deficit score after treatment, while the cerebral infarction area and water content were measured, the histopathology of hippocampus was observed by HE staining, the expression levels of brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP) and neuron-specific enolase (NSE) in hippocampus were detected with ELISA, the apoptosis of brain neurons were measured with TUNEL, and the expression levels of caspase-3 and Bcl-2 in brain homogenate were detected with Western blotting and reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR).

Results

Compared with the sham group, the neurological deficit score, cerebral infarction area and water content increased significantly (P < 0.05) in the other groups; while the the expression of GFAP and NSE in hippocampus increased (P < 0.05) and the expression of BDNF decreased (P < 0.05); the number of apoptosis in hippocampus increased (P < 0.05), and the expression of caspase-3 increased (P < 0.05) and the expression of Bcl-2 decreased (P < 0.05). Compared with the model group and non-acupoint group, the neurological deficit score, cerebral infarction area and water content decreased in the Fengchi group (P < 0.05); while the the expression of GFAP and NSE in hippocampus decreased (P < 0.05) and the expression of BDNF increased (P < 0.05); the number of apoptosis in hippocampus decreased (P < 0.05), and the expression of caspase-3 decreased (P < 0.05) and the expression of Bcl-2 increased (P < 0.05).

Conclusion

Electroacupuncture at Fengchi can protect the brain from acute cerebral infarction, adjust the expression of GFAP, NSE and BDNF, and reduce the neuronal apoptosis.

Key words: cerebral infarction, electroacupuncture, astrocyte, neurons, rats

中图分类号:

R743.32

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.

Qiong-fen WANG,Feng-bo WANG,Ke WANG,Yong-qiang ZHONG,Jiao-jiao WANG. Effect of Electroacupuncture at Fengchi on Astrocytes and Neurons in Rats with Acute Cerebral Infarction[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(3): 302-309.

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组别	n	梗死面积	脑组织含水量
假手术组	4	0	68.68±4.64
模型组	4	47.51±2.20^a	88.28±6.16^a
非穴组	4	46.84±3.10^a	86.73±5.67^a
风池组	4	24.60±2.12^a,b,c	77.30±4.51^a,b,c
统计值		H = 12.918	F = 11.851
P值		0.005	0.001

组别	n	BDNF	GFAP	NSE
假手术组	8	60.59±4.18	293.94±12.28	78.88±8.07
模型组	8	18.25±1.90^a	414.66±18.32^a	154.11±10.69^a
非穴组	8	18.19±2.18^a	413.36±15.73^a	154.43±9.93^a
风池组	8	41.55±2.75^a,b,c	323.48±14.88^a,b,c	97.31±5.74^a,b,c
F值		402.917	128.713	135.543
P值		< 0.001	< 0.001	< 0.001

组别	n	凋亡细胞数	caspase-3 mRNA (2^-ΔΔCt)	Bcl-2 mRNA (2^-ΔΔCt)	caspase-3蛋白 (/GAPDH)	Bcl-2蛋白 (/GAPDH)
假手术组	8	12.00±5.89	0.96±0.20	4.77±0.16	0.11±0.09	0.80±0.11
模型组	8	62.38±12.91^a	7.33±0.98^a	2.46±0.31^a	1.01±0.09^a	0.30±0.10^a
非穴组	8	72.13±10.30^a	7.43±0.74^a	2.91±0.37^a	1.00±0.11^a	0.31±0.10^a
风池组	8	32.63±5.10^a,b,c	4.07±0.67^a,b,c	6.98±0.27^a,b,c	0.37±0.13^a,b,c	2.01±0.10^a,b,c
F值		74.437	57.263	151.894	58.433	194.096
P值		< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

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