川芎嗪对创伤性颅脑损伤大鼠的保护作用及机制研究

易润青 全向阳

【摘要】 目的：觀察川芎嗪对创伤性颅脑损伤的保护作用及其机制。方法：选取72只雄性SD大鼠，采用PCI3000建立创伤性颅脑损伤模型，造模成功后随机分为模型组（n=15）、川芎嗪低剂量（10 mg/kg）组（n=15）、川芎嗪中剂量（20 mg/kg）组（n=15）、川芎嗪高剂量（40 mg/kg）组（n=15），同时建立假手术组（n=12）作为对照。川芎嗪三个剂量组分别尾静脉注射给予相应剂量的盐酸川芎嗪注射液，1次/d，连续7 d，模型组和假手术组给予等体积的生理盐水。给药第2、4、6、8天分别进行转棒试验和纸条粘附试验，第8天大鼠安乐死后取材进行NeuN神经元免疫组化染色和凋亡相关蛋白检测。结果：川芎嗪中、高剂量组大鼠转棒时间和纸条撕掉时间均短于模型组，差异均有统计学意义（P<0.05）;川芎嗪中、高剂量组脑梗死体积分别为（16.50±2.59）、（14.10±2.29）mm3，与模型组比较，差异均有统计学意义（P<0.05）。免疫组化染色发现川芎嗪高剂量组和模型组TBI大鼠损伤区周围皮层神经元细胞存活数目分别为（290.70±41.19）和（225.00±59.44）个，两组比较差异有统计学意义（P<0.05）;Westernblot研究可见，与模型组相比，川芎嗪高剂量组Bax蛋白表达降低、Bcl-2蛋白表达升高、Bcl-2/Bax比例升高，差异均有统计学意义（P<0.05）。结论：川芎嗪对创伤性颅脑损伤大鼠具有很好的保护作用，其机制可能与其抑制神经细胞凋亡保护神经细胞损伤有关。

【关键词】 川芎嗪; 创伤性颅脑损伤; 氧化应激; 细胞凋亡

【Abstract】 Objective：To observe the protective effect and mechanism of tetramethylpyrazine on traumatic craniocerebral injury.Method：72 male SD rats were selected and PCI3000 was used to establish the traumatic craniocerebral injury model，after successful modeling，they were randomly divided into the model group（n=15），the low dose（10 mg/kg）group（n=15），the medium dose（20 mg/kg）group（n=15），and the high dose（40 mg/kg）group（n=15），meanwhile，the sham operation group（n=12）was established as the control.Tetramethylpyrazine Hydrochloride Injection was injected into the tail vein of three dose groups，1 time/day for 7 days.The model group and the sham group were given equal volume saline.Rod transfer test and strip adhesion test were performed on days 2，4，6 and 8 after administration，after the rats were euthanized on day 8，NeuN neuron immunohistochemical staining and apoptosis-related protein detection were performed.Result：The rotarod test time and adhesive paper removal time in the tetramethylpyrazine medium group and the high dose group were significantly shorter than those in TBI model group，the differences were statistically significant（P<0.05）.Brain infarction volume was（16.50±2.59）mm3  and（14.10±2.29）mm3 in the tetramethylpyrazine medium dose group and high dose group，which were significantly decreased compared with model group（P<0.05）.Immunohistochemistry study showed that the NeuN+ neurons in the periphery area of TBI injury were（290.70±41.19）and（225.00±59.44）in the tetramethylpyrazine high dose group and model group，the difference between the two groups was statistically significant（P<0.05）.Western blot results displayed that，in tetramethylpyrazine high dose group，the Bax protein expression markedly decreased，Bcl-2 protein expression and the ratio of Bcl-2/Bax also obviously increased compared with than those of the model group（P<0.05）.Conclusion：Tetramethylpyrazine has a good protective effect on rats with traumatic brain injury and its mechanism may be related to the inhibition of apoptosis and the protection of nerve cell injury，Tetramethylpyrazine has a protective effect on rats with traumatic brain injury，which may be related to its inhibition of apoptosis.

【Key words】 Tetramethylpyrazine; Traumatic brain injury; Oxidative stress; Apoptosis

First-authors address：Guangzhou Medical Vocationl School，Guangzhou 510430，China

doi：10.3969/j.issn.1674-4985.2019.17.007

創伤性颅脑损伤（traumatic brain injury，TBI）是神经外科常见疾病。全球患病人数约为3 000多万，每年死亡人数高于100万。在中国，每年约有100万TBI患者，死亡人数近十万余人。TBI治疗除了手术和常规对症治疗外，尚无有效的治疗手段。TBI发病机制主要为直接创伤性颅脑损伤和继发的氧化应激损伤、细胞钙超载、血管应激性痉挛、氨基酸兴奋性毒性等[1]，导致神经细胞死亡、神经传递功能障碍并最终引起神经功能的缺损。由于TBI发病机制复杂，单一靶点药物药效有限，而多个靶点的药物可能对TBI治疗具有很好的作用。川芎嗪（tetramethylpyrazine，TMP）是中药川芎的有效成分，其作用机制主要包括抑制钙超载、清除自由基、舒张血管、抑制细胞凋亡、改善线粒体功能、抑制细胞炎性反应等多种作用，临床上主要用于缺血性脑卒中的治疗[2]。……