NOGO-A在创伤性脑损伤后大鼠脑组织含量表达及干预实验研究

doi:10.3877/cma.j.issn.2095-9141.2022.01.002

中华神经创伤外科电子杂志 ›› 2022, Vol. 08 ›› Issue (01) : 6 -10. doi: 10.3877/cma.j.issn.2095-9141.2022.01.002

基础研究

NOGO-A在创伤性脑损伤后大鼠脑组织含量表达及干预实验研究

崔刚¹, 王欢², 付茂武¹, 王莹¹, 段虎斌²^,^†(

)

^1. 250000　济南，山东中医药大学附属医院神经外科
^2. 030001　太原，山西医科大学第一医院神经外科

收稿日期:2021-10-28 出版日期:2022-02-15
通信作者: 段虎斌
基金资助:
国家自然科学基金(30600637); 中国博士后科学基金(2014M561207)

Experimental research of expression and intervention of NOGO-A in brain tissue of rats after traumatic brain injury

Gang Cui¹, Huan Wang², Maowu Fu¹, Ying Wang¹, Hubin Duan²^,^†()

^1. Department of Neurosurgery, The Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Ji’nan 250000, China
^2. Department of Neurosurgery, First Hospital of Shanxi Medical University, Taiyuan 030001, China

Received:2021-10-28 Published:2022-02-15
Corresponding author: Hubin Duan

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目的

研究创伤性脑损伤（TBI）后大鼠脑组织内NOGO-A分子含量及大鼠脑超微结构的改变以及二者与RHOA/ROCK信号传导通道的联系。

方法

45只健康SD大鼠按照随机数字表法分为对照组、中度创伤组和干预组，每组15只。中度创伤组和干预组大鼠采用击锤、撞杆自由落体原理致伤，制作TBI大鼠模型，干预组大鼠创伤后立即给予静脉注射盐酸法舒地尔注射液。TBI后24 h处死各组大鼠，采用电子显微镜观察各组损伤区脑组织细胞超微结构的变化，采用免疫组化技术观察各组大鼠脑组织中NOGO-A蛋白的含量，并测定NOGO-A蛋白的灰度值。

结果

TBI后24 h，中度创伤组及干预组大鼠脑组织损伤区出现病理性损伤，干预组程度较中度创伤组轻。3组大鼠脑组织内NOGO-A蛋白表达量比较，差异有统计学意义（F=176.085，P<0.05），中度创伤组的NOGO-A蛋白含量最高，干预组次之，对照组最低。

结论

TBI后24 h大鼠脑内损伤区出现细胞核、线粒体损害及细胞水肿，NOGO-A含量增加。RHOA/ROCK信号通路被阻断后能够改善TBI大鼠脑组织损伤区细胞超微结构损害以及减少NOGO-A的含量。

关键词: RHOA/ROCK信号通路, 创伤性脑损伤, 神经细胞亚结构, NOGO-A

Objective

To study the changes of the content of the NOGO-A and the ultrastructure in rat brain tissue after traumatic brain injury (TBI), and their relationship with RHOA/ROCK transduction pathway.

Methods

Forty-five healthy SD rats were randomly divided into control group, moderate trauma group and intervention group, with 15 rats in each group. The TBI rat model was established by hammer and free falling rod in moderate trauma group and intervention group. The rats in the intervention group were given fasudil hydrochloride injection intravenously immediately after trauma. The rats in each group were sacrificed 24 h after TBI. The ultrastructural changes of brain tissue cells in the injured area of each group were observed by electron microscope, the content of NOGO-A protein in brain tissue of each group was observed by immunohistochemical technique, and the gray value of NOGO-A protein was measured.

Results

Twenty-four hours after TBI, pathological injury occurred in the brain tissue injury area of rats in the moderate trauma group and the intervention group, and the intervention group was less severe than in the moderate trauma group. There was significant difference in the expression of NOGO-A protein in the brain tissue of the three groups (F=176.085, P<0.05). The moderate trauma group had the highest NOGO-A protein content, followed by the intervention group and the lowest in the control group.

Conclusion

Twenty-four hours after TBI, the injure of cell nucleus、mitochondria and cells edema appeared in the damage area of brains of rats, the distribution and the content of the NOGO-A in the damage area of rats increased. Blocking of the RHO/ROCK signaling pathway can improve the damage of the ultrastructure of the brain cell in the damage area and reduce the content and distribution of NOGO-A.

Key words: RHOA/ROCK signaling pathway, Traumatic brain injury, Ultrastructure of nerve cells, NOGO-A

图1 大鼠撞击骨窗图示

图2 电子显微镜下各组大鼠损伤区超微结构变化（×12 000）A：对照组；B：中度创伤组；C：干预组

图3 颅脑损伤后24 h各组大鼠NOGO-A蛋白的表达（免疫组化染色，×400）A：对照组；B：中度创伤组；C：干预组

表1 创伤性脑损伤后24 h的大鼠NOGO-A蛋白表达变化

组别	只数	NOGO-A蛋白表达量
对照组	15	100.10±8.25
干预组	15	127.44±9.48^a
中度创伤组	15	157.92±7.47^ab
F值		176.085
P值		<0.001

表1 创伤性脑损伤后24 h的大鼠NOGO-A蛋白表达变化

组别	只数	NOGO-A蛋白表达量
对照组	15	100.10±8.25
干预组	15	127.44±9.48^a
中度创伤组	15	157.92±7.47^ab
F值		176.085
P值		<0.001

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