RHO/ROCK信号通路对创伤性脑损伤后颅内神经系统微环境的影响

doi:10.3877/cma.j.issn.2095-9141.2022.04.003

目的

探讨创伤性脑损伤（TBI）后颅内RHO/ROCK信号通路对神经系统微环境的影响。

方法

选用健康SD大鼠195只，采用击锤、撞杆自由落体原理致伤制作脑创伤大鼠模型，作为创伤组；伤后给予部分大鼠尾静脉注射RHO/ROCK信号通路阻断剂，作为干预组；选用健康大鼠作为对照组。创伤组和干预组分别于0.5、6、12、24、48、72 h处死大鼠，每个时间点15只；对照组选取15只。各时间点取大鼠创伤区及对照组大鼠脑组织测定ROCKⅡ、NOGO-A、NF-κB蛋白的含量并进行统计学分析。

结果

创伤组大鼠脑创伤区及周围组织中ROCKⅡ、NOGO-A、NF-κB含量较对照组增多，且其在脑内的含量与时间有相关性。给予RHO/ROCK信号通路阻断剂后，干预组各时间点的ROCKⅡ、NOGO-A、NF-κB蛋白含量明显减少，差异均具有统计学意义（P<0.05）。

结论

RHO/ROCK信号通路可改变TBI后大鼠脑组织内神经系统微环境。

关键词: 创伤性脑损伤, RHO/ROCK信号通路, 神经系统微环境, ROCKⅡ, NOGO-A, 核转录因子κB

Objective

To investigate the effect of RHO/ROCK signal pathway on nervous system microenvironment after traumatic brain injury (TBI) in rats.

Methods

One hundred and ninety-five healthy SD rats were selected to make a brain trauma rat model by using the principle of hammer and rod free fall as the trauma group; After injury, some rats were injected with RHO/ROCK signal pathway blocker through caudal vein as the intervention group; Healthy rats were selected as the control group. The rats in the trauma group and the intervention group were killed at 0.5, 6, 12, 24, 48 and 72 h respectively, with 15 rats per time point; 15 rats were selected as control group. At each time point, the brain tissues of rats in the traumatic group and the control group was taken to measure the protein content of ROCK Ⅱ, NOGO-A and NF-κB.

Results

After TBI, the content of ROCK II, NOGO-A and NF-κB in brain trauma area and surrounding tissue increased in the traumatic group compared with the control group, and its content in the brain has a correlation with the time. After the administration of RHO/ROCK signaling pathway blocker, the protein content of ROCK Ⅱ, NOGO-A and NF-κB was reduced in the intervention group at each time point, and the difference were statistically significant (P<0.05).

Conclusion

RHO/ROCK signaling pathway can change the microenvironment of nervous system in rat brain after TBI.

Key words: Traumatic brain injury, RHO/ROCK signaling pathway, Nervous system microenvironment, ROCKⅡ, NOGO-A, Nuclear factor kappa B

图1 创伤标本制作过程示意图A：撞击骨窗图示（直径5 mm）；B：自由落体致伤原理图示；C：致伤完成缝合伤口图示

表1 3组大鼠脑组织ROCKⅡ蛋白含量比较

组别	0.5 h	6h	12 h	24 h	48 h	72 h	F值	P值
对照组	1.000±0.037	1.000±0.037	1.000±0.037	1.000±0.037	1.000±0.037	1.000±0.037
创伤组	1.064±0.081	1.121±0.073^a	1.180±0.044^a	1.235±0.083^a	1.288±0.072^a	1.358±0.103^a	28.990	<0.001
干预组	0.985±0.070^b	1.034±0.073^b	1.077±0.043^ab	1.108±0.069^ab	1.165±0.052^ab	1.256±0.053^ab	37.590	<0.001
F值	6.216	14.510	70.590	47.970	102.400	104.100
P值	0.004	<0.001	<0.001	<0.001	<0.001	<0.001

表1 3组大鼠脑组织ROCKⅡ蛋白含量比较

组别	0.5 h	6h	12 h	24 h	48 h	72 h	F值	P值
对照组	1.000±0.037	1.000±0.037	1.000±0.037	1.000±0.037	1.000±0.037	1.000±0.037
创伤组	1.064±0.081	1.121±0.073^a	1.180±0.044^a	1.235±0.083^a	1.288±0.072^a	1.358±0.103^a	28.990	<0.001
干预组	0.985±0.070^b	1.034±0.073^b	1.077±0.043^ab	1.108±0.069^ab	1.165±0.052^ab	1.256±0.053^ab	37.590	<0.001
F值	6.216	14.510	70.590	47.970	102.400	104.100
P值	0.004	<0.001	<0.001	<0.001	<0.001	<0.001

表2 3组大鼠脑组织NOGO-A蛋白含量比较

组别	0.5 h	6 h	12 h	24 h	48 h	72 h	F值	P值
对照组	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082
创伤组	1.078±0.153	1.337±0.118^a	1.407±0.133^a	1.578±0.075^a	1.590±0.077^a	1.692±0.058^a	63.810	<0.001
干预组	1.043±0.088	1.099±0.090^ab	1.217±0.095^ab	1.273±0.095^ab	1.427±0.048^ab	1.562±0.035^ab	92.960	<0.001
F值	1.814	46.590	55.930	176.300	276.800	531.400
P值	0.175	<0.001	<0.001	<0.001	<0.001	<0.001

表2 3组大鼠脑组织NOGO-A蛋白含量比较

组别	0.5 h	6 h	12 h	24 h	48 h	72 h	F值	P值
对照组	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082
创伤组	1.078±0.153	1.337±0.118^a	1.407±0.133^a	1.578±0.075^a	1.590±0.077^a	1.692±0.058^a	63.810	<0.001
干预组	1.043±0.088	1.099±0.090^ab	1.217±0.095^ab	1.273±0.095^ab	1.427±0.048^ab	1.562±0.035^ab	92.960	<0.001
F值	1.814	46.590	55.930	176.300	276.800	531.400
P值	0.175	<0.001	<0.001	<0.001	<0.001	<0.001

表3 3组大鼠脑组织NF-κB蛋白含量比较

组别	0.5 h	6h	12 h	24 h	48 h	72 h	F值	P值
对照组	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082
创伤组	1.113±0.047^a	1.161±0.068^a	1.220±0.057^a	1.300±0.054^a	1.350±0.060^a	1.428±0.065^a	61.650	<0.001
干预组	1.046±0.103	1.093±0.083^a	1.130±0.066^ab	1.157±0.071^ab	1.265±0.090^ab	1.336±0.070^ab	26.800	<0.001
F值	2.435	4.328	38.270	68.550	80.860	142.900
P值	0.171	0.075	<0.001	<0.001	<0.001	<0.001

表3 3组大鼠脑组织NF-κB蛋白含量比较

组别	0.5 h	6h	12 h	24 h	48 h	72 h	F值	P值
对照组	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082	1.000±0.082
创伤组	1.113±0.047^a	1.161±0.068^a	1.220±0.057^a	1.300±0.054^a	1.350±0.060^a	1.428±0.065^a	61.650	<0.001
干预组	1.046±0.103	1.093±0.083^a	1.130±0.066^ab	1.157±0.071^ab	1.265±0.090^ab	1.336±0.070^ab	26.800	<0.001
F值	2.435	4.328	38.270	68.550	80.860	142.900
P值	0.171	0.075	<0.001	<0.001	<0.001	<0.001

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Effects of RHO/ROCK signaling pathway on the microenvironment of nervous system after traumatic brain injury in rats

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Effects of RHO/ROCK signaling pathway on the microenvironment of nervous system after traumatic brain injury in rats