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

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。

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

ObjectiveTo investigate the effect of Xingnaojing injection on neurological functions of rats with craniocerebral injury and its related mechanisms. MethodsA 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. ResultsThere 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). ConclusionXingnaojing 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.