早期运动干预对脑缺血大鼠脑神经髓鞘的影响及机制研究

doi:10.11958/20231795

天津医药 ›› 2024, Vol. 52 ›› Issue (6): 589-594.doi: 10.11958/20231795

早期运动干预对脑缺血大鼠脑神经髓鞘的影响及机制研究

王俊懿(), 李宸, 吴昕岳, 丁心语, 万春晓()

天津医科大学总医院康复医学科（邮编300052）

收稿日期:2023-11-20 修回日期:2023-12-12 出版日期:2024-06-15 发布日期:2024-06-06
通讯作者: ^△E-mail：cwan@tmu.edu.cn
作者简介:王俊懿（1999），女，硕士在读，主要从事神经康复方面研究。E-mail：orihara_0504@163.com
基金资助:
天津市医学重点学科（专科）建设项目(TJYXZDXK-060B);天津市卫生健康科技项目重点学科专项(TJWJ2022XK007);天津市应用基础研究多元投入基金面上项目(21JCYBJC01610)

Effect and mechanism of early exercise intervention on cerebral nerve myelin in rats with cerebral ischemia

WANG Junyi(), LI Chen, WU Xinyue, DING Xinyu, WAN Chunxiao()

Department of Physical Medicine and Rehabilitation, Tianjin Medical University General Hospital, Tianjin 300052, China

Received:2023-11-20 Revised:2023-12-12 Published:2024-06-15 Online:2024-06-06
Contact: ^△E-mail: cwan@tmu.edu.cn

王俊懿, 李宸, 吴昕岳, 丁心语, 万春晓. 早期运动干预对脑缺血大鼠脑神经髓鞘的影响及机制研究[J]. 天津医药, 2024, 52(6): 589-594.

WANG Junyi, LI Chen, WU Xinyue, DING Xinyu, WAN Chunxiao. Effect and mechanism of early exercise intervention on cerebral nerve myelin in rats with cerebral ischemia[J]. Tianjin Medical Journal, 2024, 52(6): 589-594.

摘要/Abstract

摘要：

目的探讨早期运动干预通过抑制内质网应激诱导的细胞凋亡对脑缺血大鼠脑神经髓鞘损伤的影响。方法 18只SD大鼠随机分为假手术（SHAM）组，大脑中动脉闭塞静息（MCAO-SED）组、大脑中动脉闭塞运动（MCAO-EX）组，每组6只。除SHAM组外，其他各组采用改进的Longa线栓塞法制备大脑中动脉闭塞模型。造模后MCAO-EX组大鼠置于跑步机上进行运动干预28 d。采用改良的神经功能缺损评分（mNSS）评估神经功能，MRI扫描（T2）检测脑梗死体积，免疫荧光染色检测髓磷脂碱性蛋白（MBP）表达，透射电子显微镜观察髓鞘的结构，Western blot法检测内质网应激相关蛋白表达，TUNEL染色检测细胞凋亡。结果干预28 d后，同MCAO-SED组比，MCAO-EX组神经功能恢复良好，梗死体积减少，髓鞘完整性增加，MBP荧光强度表达增加，MBP的表达水平增加，同时转录激活因子6（ATF6）、磷酸化肌醇需求酶1（p-IRE1）、磷酸化蛋白激酶R样内质网激酶（p-PERK）、活化的胱天蛋白酶-3（cleaved caspase 3）蛋白的表达水平显著降低，细胞凋亡减少。结论早期运动可抑制内质网应激诱导的细胞凋亡，促进脑神经髓鞘修复，减少梗死面积，改善神经功能。

关键词: 脑缺血, 髓鞘, 内质网应激, 凋亡, 早期运动干预

Abstract:

Objective To investigate the effect and potential mechanism of early exercise intervention on cerebral myelin in cerebral ischemia rats. Methods A total of 18 SD rats were randomly divided into the sham group, the middle cerebral artery occlusion resting group (MCAO-SED) and the middle cerebral artery occlusion exercise group (MCAO-EX), with 6 rats in each group. Except the sham group, the middle cerebral artery occlusion model was prepared by modified Longa line embolization method in other groups. After modeling, rats in the MCAO-EX group were placed on a treadmill for exercise intervention for 28 days. Neurological function was assessed by modified neural function deficit score (mNSS). Infarct volume was detected by MRI scanning (T2), myelin basic protein (MBP) expression was detected by immunofluorescence. Myelin sheath structure was detected by transmission electron microscopy. Western blot assay was used to detect endoplasmic reticulum stress-related protein expression. Apoptosis was detected by TUNEL staining. Results After 28 days of intervention, compared with the MCAO-SED group, the nerve function recovered well in the MCAO-EX group, infarct volume decreased, myelin integrity increased, MBP fluorescence intensity expression increased and MBP expression level increased. The expression levels of ATF6, p-IRE1, p-PERK and cleaved caspase 3 were significantly decreased, and apoptosis was reduced. Conclusion Early exercise can inhibit endoplasmic reticulum stress-induced apoptosis, promote cerebral myelin repair, reduce infarct size and improve nerve function.

Key words: brain ischemia, myelin sheath, endoplasmic reticulum stress, apoptosis, early exercise intervention

中图分类号:

R743.33

图/表 9

表1 各组大鼠mNSS评分比较（n=6，分，$\bar{x}±s$）

Tab.1 Comparison of mNSS scores between three groups of rats

组别	第1天			第3天		第7天
SHAM组	1.66±0.81			0.17±0.41		0.00±0.00
MCAO-SED组	12.33±1.86^a			9.50±2.26^a		8.33±1.21^a
MCAO-EX组	12.17±1.60			8.50±1.38		6.33±1.03^b
组别		第14天	第21天		第28天
SHAM组		0.00±0.00	0.00±0.00		0.00±0.00
MCAO-SED组		6.17±1.33^a	5.17±0.98^a		4.67±0.82^a
MCAO-EX组		4.33±1.03^b	3.67±0.82^b		3.33±0.52^b

图1 干预前大鼠的脑T2WI成像

Fig.1 T2WI imaging of rats before intervention

图2 干预第28天大鼠的脑T2WI成像

Fig.2 T2WI imaging of rats 28 days after intervention

表2 各组大鼠脑梗死体积比变化比较（%，$\bar{x}±s$）

Tab.2 Comparison of changes of cerebral infarction volume ratio between three groups

组别	n	干预前	干预第28天
SHAM组	3	0.00±0.00	0.00±0.00
MCAO-SED组	3	41.23±0.87^a	30.36±1.40^a
MCAO-EX组	3	41.42±1.13	22.04±0.21^b
F		0.053	102.521^＊＊

图3 透射电镜下各组干预第28天外囊髓鞘完整性超微结构（×6 000）

Fig.3 Ultrastructure of myelin integrity of the external capsule indicated by transmission electron microscope at 28 days of intervention in each group (×6 000)

图4 各组大鼠脑组织MBP免疫荧光染色（×200）绿色荧光为MBP，蓝色荧光为DAPI。

Fig.4 MBP immunofluorescence staining of brain tissue in each group (×200)

图5 各组大鼠ATF6、IRE1、PERK、p-IRE1、p-PERK、cleaved caspase 3、MBP蛋白表达水平 A：SHAM组；B：MCAO-SED组；C：MCAO-EX组。

Fig.5 Expression levels of ATF6, IRE1, PERK, p-IRE1, p-PERK, cleaved caspase 3 and MBP in each group

表3 各组大鼠ATF6、IRE、PERK、p-IRE、p-PERK、cleaved caspase 3及MBP蛋白表达比较（n=4，$\bar{x}±s$）

Tab.3 Comparison of ATF6, IRE, PERK, p-IRE, p-PERK, cleaved caspase 3 and MBP protein expression between three groups of rats

组别	ATF6	IRE1	p-IRE1	PERK	p-PERK	cleaved caspase 3	MBP
SHAM组	1.00±0.25	1.00±0.23	1.00±0.33	1.00±0.17	1.00±0.11	1.00±0.09	1.00±0.09
MCAO-SED组	2.16±0.33^a	1.05±0.13	1.96±0.20^a	0.95±0.19	1.92±0.27^a	2.86±0.88^a	0.50±0.08^a
MCAO-EX组	1.45±0.02^b	1.05±0.17	1.45±0.27^b	1.02±0.13	1.15±0.28^b	1.60±0.45^b	0.75±0.19^b
F	24.841^＊＊	0.091	12.710^＊＊	0.163	17.6743^＊＊	50.112^＊＊	13.867^＊＊

图6 各组大鼠脑组织TUNEL染色（×200）绿色荧光为TUNEL阳性染色，蓝色荧光为DAPI染色。

Fig.6 TUNEL staining of brain tissue of rats in each group (×200)

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早期运动干预对脑缺血大鼠脑神经髓鞘的影响及机制研究

Effect and mechanism of early exercise intervention on cerebral nerve myelin in rats with cerebral ischemia

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