醒脑静注射液对颅脑损伤大鼠神经功能的影响及机制研究

doi:10.3877/cma.j.issn.1674-6880.2018.01.005

中华危重症医学杂志(电子版) ›› 2018, Vol. 11 ›› Issue (01) : 29 -34. doi: 10.3877/cma.j.issn.1674-6880.2018.01.005

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论著

醒脑静注射液对颅脑损伤大鼠神经功能的影响及机制研究

陈亦华¹^,^†(

), 程振宇¹, 邵林华¹, 何忠平², 王瑞权³, 傅斌⁴, 杜骁龙⁵

^1. 321000　浙江金华，金华市人民医院神经外科
^2. 321000　浙江金华，金华市食品药品检验检测研究院药理所
^3. 321000　浙江金华，金华市人民医院病理科
^5. 321000　浙江金华，金华市人民医院外科

收稿日期:2017-06-22 出版日期:2018-02-01
通信作者: 陈亦华
基金资助:
金华市科技计划项目(2015-3-039)

Protective effect of Xingnaojing injection on neurological functions of rats with craniocerebral injury and its related mechanisms

Yihua Chen¹^,^†(), Zhenyu Cheng¹, Linhua Shao¹, Zhongping He², Ruiquan Wang³, Bin Fu⁴, Xiaolong Du⁵

^1. Department of Neurosurgery, Jinhua People's Hospital, Jinhua 321000, China
^2. Institute of Pathology, Jinhua Institute for Food and Drug Control, Jinhua 321000, China
^3. Department of Pathology, Jinhua People's Hospital, Jinhua 321000, China
^4. Department of Surgery, Jinhua People's Hospital, Jinhua 321000, China

Received:2017-06-22 Published:2018-02-01
Corresponding author: Yihua Chen
About author:
Corresponding author: Chen Yihua, Email: rmyycyh@163.com

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引用本文：

陈亦华, 程振宇, 邵林华, 何忠平, 王瑞权, 傅斌, 杜骁龙. 醒脑静注射液对颅脑损伤大鼠神经功能的影响及机制研究[J/OL]. 中华危重症医学杂志(电子版), 2018, 11(01): 29-34.

Yihua Chen, Zhenyu Cheng, Linhua Shao, Zhongping He, Ruiquan Wang, Bin Fu, Xiaolong Du. Protective effect of Xingnaojing injection on neurological functions of rats with craniocerebral injury and its related mechanisms[J/OL]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2018, 11(01): 29-34.

目的

探讨醒脑静注射液对颅脑损伤大鼠神经功能的影响及相关机制。

方法

108只大鼠采用随机数字表法分成假手术组、创伤组和治疗组，每组36只。采用改进的Feeney自由落体损伤装置制作大鼠颅脑损伤模型，假手术组和创伤组大鼠腹腔注射1 mL等渗NaCl溶液，治疗组大鼠按5 mL/kg的剂量腹腔注射醒脑静注射液，1次/d，连续7 d。每组6只大鼠用于神经功能检测，另外每组分别在12、24、48、72和168 h时点断头处死6只大鼠，分离海马组织。采用Western-blotting检测Survivin蛋白表达，末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法（TUNEL）检测神经细胞凋亡情况。采用前肢放置试验评分和平衡实验评分对术后7、14、21和28 d大鼠进行行为测试，改良神经功能缺损程度评分测定各组大鼠神经功能损伤情况。

结果

三组间大鼠海马组织Survivin蛋白表达和神经细胞凋亡比例比较，差异有统计学意义（F=262.495、203.219，P均< 0.05）。12、24、48、72和168 h创伤组及治疗组大鼠海马组织Survivin蛋白表达和神经细胞凋亡比例均较同时间点假手术组显著升高（P均< 0.05）；与同时间点创伤组比较，48、72和168 h治疗组大鼠海马组织Survivin蛋白表达均显著升高（P均< 0.05），而48、72和168 h神经细胞凋亡比例均较同时间点创伤组显著下降。三组前肢放置试验评分和平衡实验评分比较，差异均有统计学意义（F=60.876、23.143，P均< 0.05）。术后7、14、21和28 d治疗组大鼠前肢放置试验评分和平衡实验评分均较同时间点创伤组显著下降（P均< 0.05）。三组大鼠改良神经功能缺损程度评分比较，差异有统计学意义（F=24.046，P < 0.001）。术后7、14、21和28 d治疗组大鼠改良神经功能缺损程度评分均较同时间点创伤组显著下降（P均< 0.05）。

结论

醒脑静注射液可能通过上调Survivin蛋白表达抑制神经细胞凋亡，从而发挥保护颅脑损伤大鼠神经功能的作用。

关键词: 醒脑静注射液, 大鼠, 神经功能, 神经细胞凋亡, Survivin

Objective

To investigate the effect of Xingnaojing injection on neurological functions of rats with craniocerebral injury and its related mechanisms.

Methods

A total of 108 rats were randomly divided into the sham operation group, trauma group and treatment group, each with 36 rats. The craniocerebral injury model was established using the improved Feeney free fall injury device. Rats in the sham operation and trauma groups were injected intraperitoneally with 1 mL normal saline, while rats in the treatment group were administrated intraperitoneally with 5 mL/kg Xingnaojing injection, once a day for 7 days. Six rats in each group were used for neurological function tests. At 12, 24, 48, 72, and 168 h, 6 rats in each group were sacrificed to separate hippocampal tissues, respectively. The Western-blotting method was used to detect the expression of Survivin, and terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining was used to detect neuronal apoptosis. The forelimb placement test and balanced experiment were used to test behaviors of rats, and the degree of improved neurologic impairment was used to measure neurological injury of rats in each group on 7, 14, 21, and 28 d after operation.

Results

There were significant differences in the expressions of Survivin and neuron apoptosis among the three groups (F=262.495, 203.219, both P < 0.05). The proportions of Survivin and neuronal apoptosis in hippocampus were significantly higher in the trauma and treatment groups than in the sham operation group at 12, 24, 48, 72, and 168 h (all P < 0.05). Compared with the trauma group at 48, 72, and 168 h, the expression of Survivin significantly increased and the percentage of neuron apoptosis significantly declined in the treatment group (all P < 0.05). The scores of forelimb placement test and balanced experiment in the three groups were statistically significant (F=60.876, 23.143; both P < 0.05). The scores on 7, 14, 21, and 28 d after operation were significantly lower in the treatment group than in the trauma group (all P < 0.05). The scores of improved neurologic impairment degree in the three groups were also statistically significant (F=24.046, P < 0.001). The scores on 7, 14, 21, and 28 d after operation were significantly lower in the treatment group than in the trauma group (all P < 0.05).

Conclusion

Xingnaojing injection may exert a protective effect on neurological functions of craniocerebral injury rats by up-regulating the expression of Survivin and subsequently inhibiting neuronal apoptosis.

Key words: Xingnaojing injection, Rats, Neurological function, Neuronal apoptosis, Survivin

表1 三组颅脑损伤大鼠各时间点海马组织Survivin蛋白表达的变化（±s）

组别	只数	12 h	24 h	48 h	72 h	168 h
假手术组	30	100.0 ± 14.5	97.7 ± 12.8	108.5 ± 15.2	102.3 ± 15.9	102.2 ± 15.7
创伤组	30	213.7 ± 34.8^a	282.0 ± 35.3^a	454.1 ± 72.5^a	398.4 ± 44.7^a	288.9 ± 26.4^a
治疗组	30	224.5 ± 32.0^a	313.5 ± 32.7^a	582.8 ± 61.3^ab	498.2 ± 72.0^ab	360.9 ± 30.0^ab

表1 三组颅脑损伤大鼠各时间点海马组织Survivin蛋白表达的变化（±s）

组别	只数	12 h	24 h	48 h	72 h	168 h
假手术组	30	100.0 ± 14.5	97.7 ± 12.8	108.5 ± 15.2	102.3 ± 15.9	102.2 ± 15.7
创伤组	30	213.7 ± 34.8^a	282.0 ± 35.3^a	454.1 ± 72.5^a	398.4 ± 44.7^a	288.9 ± 26.4^a
治疗组	30	224.5 ± 32.0^a	313.5 ± 32.7^a	582.8 ± 61.3^ab	498.2 ± 72.0^ab	360.9 ± 30.0^ab

表2 三组颅脑损伤大鼠各时间点海马神经细胞凋亡比例的变化（%，±s）

组别	只数	12 h	24 h	48 h	72 h	168 h
假手术组	30	9.2 ± 2.1	10.5 ± 1.9	11.2 ± 2.9	10.2 ± 2.0	10.1 ± 1.9
创伤组	30	35.3 ± 4.3^a	54.6 ± 7.0^a	50.8 ± 6.3^a	44.2 ± 4.4^a	30.9 ± 4.5^a
治疗组	30	33.8 ± 4.9^a	48.9 ± 6.4^a	41.8 ± 6.6^ab	37.7 ± 2.2^ab	25.4 ± 3.1^ab

表2 三组颅脑损伤大鼠各时间点海马神经细胞凋亡比例的变化（%，±s）

组别	只数	12 h	24 h	48 h	72 h	168 h
假手术组	30	9.2 ± 2.1	10.5 ± 1.9	11.2 ± 2.9	10.2 ± 2.0	10.1 ± 1.9
创伤组	30	35.3 ± 4.3^a	54.6 ± 7.0^a	50.8 ± 6.3^a	44.2 ± 4.4^a	30.9 ± 4.5^a
治疗组	30	33.8 ± 4.9^a	48.9 ± 6.4^a	41.8 ± 6.6^ab	37.7 ± 2.2^ab	25.4 ± 3.1^ab

表3 颅脑损伤大鼠各时间点前肢放置实验评分的变化（分，±s）

组别	只数	1 d	3 d	7 d	14 d	21 d	28 d
创伤组	6	8.8 ± 0.4	8.0 ± 0.6	7.5 ± 0.6	6.5 ± 0.6	4.8 ± 0.8	4.0 ± 0.9
治疗组	6	8.5 ± 0.6	7.7 ± 0.5	6.5 ± 0.6	4.7 ± 0.8	3.2 ± 0.8	2.5 ± 0.6
t值	?	1.195	1.000	3.162	5.596	3.835	3.503
P值	?	0.260	0.341	0.010	0.001	0.003	0.006

表3 颅脑损伤大鼠各时间点前肢放置实验评分的变化（分，±s）

组别	只数	1 d	3 d	7 d	14 d	21 d	28 d
创伤组	6	8.8 ± 0.4	8.0 ± 0.6	7.5 ± 0.6	6.5 ± 0.6	4.8 ± 0.8	4.0 ± 0.9
治疗组	6	8.5 ± 0.6	7.7 ± 0.5	6.5 ± 0.6	4.7 ± 0.8	3.2 ± 0.8	2.5 ± 0.6
t值	?	1.195	1.000	3.162	5.596	3.835	3.503
P值	?	0.260	0.341	0.010	0.001	0.003	0.006

表4 颅脑损伤大鼠各时间点平衡实验评分的变化（分，±s）

组别	只数	1 d	3 d	7 d	14 d	21 d	28 d
创伤组	6	5.8 ± 0.8	5.3 ± 0.5	5.2 ± 0.8	4.3 ± 1.2	3.7 ± 0.5	3.0 ± 0.9
治疗组	6	5.5 ± 0.6	4.7 ± 0.8	4.0 ± 0.9	2.8 ± 0.8	2.5 ± 0.6	1.8 ± 0.4
t值	?	0.877	1.690	2.445	2.577	3.796	2.907
P值	?	0.401	0.122	0.035	0.032	0.004	0.016

表4 颅脑损伤大鼠各时间点平衡实验评分的变化（分，±s）

组别	只数	1 d	3 d	7 d	14 d	21 d	28 d
创伤组	6	5.8 ± 0.8	5.3 ± 0.5	5.2 ± 0.8	4.3 ± 1.2	3.7 ± 0.5	3.0 ± 0.9
治疗组	6	5.5 ± 0.6	4.7 ± 0.8	4.0 ± 0.9	2.8 ± 0.8	2.5 ± 0.6	1.8 ± 0.4
t值	?	0.877	1.690	2.445	2.577	3.796	2.907
P值	?	0.401	0.122	0.035	0.032	0.004	0.016

表5 颅脑损伤大鼠各时间点改良神经功能缺损程度评分的变化（分，±s）

组别	只数	1d	3d	7 d	14 d	21 d	28 d
创伤组	6	14.0 ± 1.3	12.5 ± 1.9	10.0 ± 1.1	8.5 ± 0.6	7.0 ± 0.6	6.0 ± 0.9
治疗组	6	13.2 ± 1.5	11.7 ± 2.3	8.0 ± 1.4	6.3 ± 1.0	5.5 ± 1.1	4.2 ± 1.0
t值	?	1.052	0.682	2.739	4.540	3.000	3.372
P值	?	0.318	0.511	0.021	0.002	0.013	0.007

表5 颅脑损伤大鼠各时间点改良神经功能缺损程度评分的变化（分，±s）

组别	只数	1d	3d	7 d	14 d	21 d	28 d
创伤组	6	14.0 ± 1.3	12.5 ± 1.9	10.0 ± 1.1	8.5 ± 0.6	7.0 ± 0.6	6.0 ± 0.9
治疗组	6	13.2 ± 1.5	11.7 ± 2.3	8.0 ± 1.4	6.3 ± 1.0	5.5 ± 1.1	4.2 ± 1.0
t值	?	1.052	0.682	2.739	4.540	3.000	3.372
P值	?	0.318	0.511	0.021	0.002	0.013	0.007

[1]	Majdan M, Brazinova A, Rusnak M, et al. Outcome prediction after traumatic brain injury: comparison of the performance of routinely used severity scores and multivariable prognostic models[J]. J Neurosci Rural Pract, 2017, 8 (1): 20-29.
[2]	王林国，俞文华，董晓巧，等.血浆和肽素浓度对重型颅脑损伤后急性创伤性凝血病的预测价值[J/CD].中华危重症医学杂志（电子版），2015，8（5）：300-305.
[3]	Di Pietro V, Ragusa M, Davies D, et al. MicroRNAs as novel biomarkers for the diagnosis and prognosis of mild and severe traumatic brain injury[J]. J Neurotrauma, 2017, 34 (11): 1948-1956.
[4]	da P Oliveira DM, Pereira CU, da P Freitas ZM. Pr-ognosis of patients with traumatic brain injury after implementation of a nurse assessment protocol[J]. J Neurosci Nurs, 2016, 48 (5): 278-284.
[5]	Babaee A, Eftekhar-Vaghefi SH, Asadi-Shekaari M, et al. Melatonin treatment reduces astrogliosis and apoptosis in rats with traumatic brain injury[J]. Iran J Basic Med Sci, 2015, 18 (9): 867-872.
[6]	Yao J, Zheng K, Zhang X. Rosiglitazone exerts neuro-protective effects via the suppression of neuronal autophagy and apoptosis in the cortex following traumatic brain injury[J]. Mol Med Rep, 2015, 12 (5): 6591-6597.
[7]	Chuang CH, Hsu YC, Wang CC, et al. Cerebral blood flow and apoptosis-associated factor with electroacupuncture in a traumatic brain injury rat model[J]. Acupunct Med, 2013, 31 (4): 395-403.
[8]	Brany D, Dvorská D, Slávik P, et al. Survivin and gynaecological tumours[J]. Pathol Res Pract, 2017, 213 (4): 295-300.
[9]	Roy K, Singh N, Kanwar RK, et al. Survivin modul-ators: an updated patent review (2011-2015)[J]. Recent Pat Anticancer Drug Discov, 2016, 11 (2): 152-169.
[10]	Chen X, Duan N, Zhang C, et al. Survivin and tumorigenesis: molecular mechanisms and therapeutic strategies[J]. J Cancer, 2016, 7 (3): 314-323.
[11]	Sah NK, Seniya C. Survivin splice variants and their diagnostic significance[J]. Tumour Biol, 2015, 36 (9): 6623-6631.
[12]	Zhang L, Yan R, Zhang Q, et al. Survivin, a key component of the Wnt/β-catenin signaling pathway, contributes to traumatic brain injury-induced adult neurogenesis in the mouse dentate gyrus[J]. Int J Mol Med, 2013, 32 (4): 867-875.
[13]	Zhang Z, Wang H, Jin Z, et al. Downregulation of survivin regulates adult hippocampal neurogenesis and apoptosis, and inhibits spatial learning and memory following traumatic brain injury[J]. Neuroscience, 2015 (300): 219-228.
[14]	Wu L, Zhang H, Xing Y, et al. Meta-analysis of the effects of Xingnaojing injection on consciousness disturbance[J]. Medicine (Baltimore), 2016, 95 (7): e2875.
[15]	Pan W, Yang L, Feng W, et al. Determination of five sesquiterpenoids in Xingnaojing injection by quantitative analysis of multiple components with a single marker[J]. J Sep Sci, 2015, 38 (19): 3313-3323.
[16]	Wei J, Yao L, Yang L, et al. Alteration of glutamate/GABA balance during acute alcohol intoxication in rats: effect of Xingnaojing injection[J]. J Ethnopharmacol,2015 (166): 333-339.
[17]	Xu M, Su W, Xu QP, et al. Effect of Xingnaojing injection on cerebral edema and blood-brain barrier in rats following traumatic brain injury[J]. Chin J Traumatol, 2010, 13 (3): 158-162.
[18]	Kawamata T, Dietrich WD, Schallert T, et al. Intraci-sternal basic fibroblast growth factor enhances functional recovery and up-regulates the expression of a molecular marker of neuronal sprouting following focal cerebral infarction[J]. Proc Natl Acad Sci USA, 1997, 94 (15): 8179-8184.
[19]	Chen J, Li Y, Wang L, et al. Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats[J]. Stroke, 2001, 32 (4): 1005-1011.
[20]	涂悦，杨细平，商崇智.醒脑静对颅脑创伤的保护作用[J].中国应用生理学杂志，2014，30（3）：230-236.
[21]	樊茉丽，周广喜.醒脑静对脑出血大鼠脑水肿及血肿周围基质金属蛋白酶-9和补体C3的影响[J].中国现代神经疾病杂志，2009，9（3）：284-289.
[22]	戴永建，戢翰升.醒脑静注射液对大鼠脑损伤后细胞凋亡及相关蛋白表达的影响[J].中国临床神经外科杂志，2006，11（9）：551-553.

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Protective effect of Xingnaojing injection on neurological functions of rats with craniocerebral injury and its related mechanisms

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Protective effect of Xingnaojing injection on neurological functions of rats with craniocerebral injury and its related mechanisms