川芎嗪对创伤性颅脑损伤大鼠的保护作用及机制研究
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摘要
目的:观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。
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.
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.
引文
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[3]宫有文,吕延锋.颅内动脉瘤性蛛网膜下腔出血血管痉挛治疗的临床探讨[J].中国实用神经疾病杂志,2015,18(18):66-67.
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[8]Lu H,Wang Y,He X,et al.Netrin-1 hyperexpression in mouse brain promotes angiogenesis and long-term neurological recovery after transient focal ischemia[J].Stroke,2012,43(3):838-843.
[9]Tezer F I,Firat A,Tuzun E,et al.Immunopathology in drug resistant mesial temporal lobe epilepsy with different types of hippocampal sclerosis[J].Int J Neurosci,2018,128(5):421-428.
[10]Shao Z,Wu P,Wang X,et al.Tetramethylpyrazine Protects Against Early Brain Injury and Inhibits the PERK/Akt Pathway in a Rat Model of Subarachnoid Hemorrhage[J].Neurochem Res,2018,43(8):1650-1659.
[11]Davanzo J R,Sieg E P,Timmons S D.Management of Traumatic Brain Injury[J].Surg Clin North Am,2017,97(6):1237-1253.
[12]O’leary R A,Nichol A D.Pathophysiology of severe traumatic brain injury[J].J Neurosurg Sci,2018,62(5):542-548.
[13]Kaur P,Sharma S.Recent Advances in Pathophysiology of Traumatic Brain Injury[J].Curr Neuropharmacol,2018,16(8):1224-1238.
[14]Chen K J,Chen K.Ischemic stroke treated with Ligusticum chuanxiong[J].Chin Med J(Engl),1992,105(10):870-873.
[15]Tsai T H,Liang C.Pharmacokinetics of tetramethylpyrazine in rat blood and brain using microdialysis[J].Int J Pharm,2001,216(1-2):61-66.
[16]Jiang Y R,Chen K J.Pharmacological roles of ligustrazine in cardio-/cerebrovascular systems and its progress in researches of clinical application[J].Zhongguo Zhong Xi Yi Jie He Za Zhi,2013,33(5):707-711.
[17]Zhao T,Fu Y,Sun H,et al.Ligustrazine suppresses neuron apoptosis via the Bax/Bcl-2 and caspase-3 pathway in PC12 cells and in rats with vascular dementia[J].IUBMB Life,2018,70(1):60-70.
[18]Liu S Y,Sylvester D M.Antithrombotic/antiplatelet activity of tetramethylpyrazine[J].Thromb Res,1990,58(2):129-140.
[19]Liu S Y,Sylvester D M.Antiplatelet activity of tetramethylpyrazine[J].Thromb Res,1994,75(1):51-62.
[20]Shen T,Xu H,Weng W,et al.Single-and multiple-dose pharmacokinetics of a novel tetramethylpyrazine reservoirtype transdermal patch versus tetramethylpyrazine phosphate oral tablets in healthy normal volunteers,and in vitro/in vivo correlation[J].Biol Pharm Bull,2013,36(6):931-937.
[2]Zhao Y,Liu Y,Chen K.Mechanisms and Clinical Application of Tetramethylpyrazine(an Interesting Natural Compound Isolated from Ligusticum Wallichii):Current Status and Perspective[J].Oxid Med Cell Longev,2016,2016:2124638.
[3]宫有文,吕延锋.颅内动脉瘤性蛛网膜下腔出血血管痉挛治疗的临床探讨[J].中国实用神经疾病杂志,2015,18(18):66-67.
[4]d'Avila J C,Lam T I,Bingham D,et al.Microglial activation induced by brain trauma is suppressed by post-injury treatment with a PARP inhibitor[J].J Neuroinflammation,2012,9:31.
[5]Liao S L,Kao T K,Chen W Y,et al.Tetramethylpyrazine reduces ischemic brain injury in rats[J].Neurosci Lett,2004,372(1-2):40-45.
[6]Hamm R J.Neurobehavioral assessment of outcome following traumatic brain injury in rats:an evaluation of selected measures[J].J Neurotrauma,2001,18(11):1207-1216.
[7]Thompson H J,Marklund N,LeBold D G,et al.Tissue sparing and functional recovery following experimental traumatic brain injury is provided by treatment with an anti-myelin-associated glycoprotein antibody[J].Eur J Neurosci,2006,24(11):3063-3072.
[8]Lu H,Wang Y,He X,et al.Netrin-1 hyperexpression in mouse brain promotes angiogenesis and long-term neurological recovery after transient focal ischemia[J].Stroke,2012,43(3):838-843.
[9]Tezer F I,Firat A,Tuzun E,et al.Immunopathology in drug resistant mesial temporal lobe epilepsy with different types of hippocampal sclerosis[J].Int J Neurosci,2018,128(5):421-428.
[10]Shao Z,Wu P,Wang X,et al.Tetramethylpyrazine Protects Against Early Brain Injury and Inhibits the PERK/Akt Pathway in a Rat Model of Subarachnoid Hemorrhage[J].Neurochem Res,2018,43(8):1650-1659.
[11]Davanzo J R,Sieg E P,Timmons S D.Management of Traumatic Brain Injury[J].Surg Clin North Am,2017,97(6):1237-1253.
[12]O’leary R A,Nichol A D.Pathophysiology of severe traumatic brain injury[J].J Neurosurg Sci,2018,62(5):542-548.
[13]Kaur P,Sharma S.Recent Advances in Pathophysiology of Traumatic Brain Injury[J].Curr Neuropharmacol,2018,16(8):1224-1238.
[14]Chen K J,Chen K.Ischemic stroke treated with Ligusticum chuanxiong[J].Chin Med J(Engl),1992,105(10):870-873.
[15]Tsai T H,Liang C.Pharmacokinetics of tetramethylpyrazine in rat blood and brain using microdialysis[J].Int J Pharm,2001,216(1-2):61-66.
[16]Jiang Y R,Chen K J.Pharmacological roles of ligustrazine in cardio-/cerebrovascular systems and its progress in researches of clinical application[J].Zhongguo Zhong Xi Yi Jie He Za Zhi,2013,33(5):707-711.
[17]Zhao T,Fu Y,Sun H,et al.Ligustrazine suppresses neuron apoptosis via the Bax/Bcl-2 and caspase-3 pathway in PC12 cells and in rats with vascular dementia[J].IUBMB Life,2018,70(1):60-70.
[18]Liu S Y,Sylvester D M.Antithrombotic/antiplatelet activity of tetramethylpyrazine[J].Thromb Res,1990,58(2):129-140.
[19]Liu S Y,Sylvester D M.Antiplatelet activity of tetramethylpyrazine[J].Thromb Res,1994,75(1):51-62.
[20]Shen T,Xu H,Weng W,et al.Single-and multiple-dose pharmacokinetics of a novel tetramethylpyrazine reservoirtype transdermal patch versus tetramethylpyrazine phosphate oral tablets in healthy normal volunteers,and in vitro/in vivo correlation[J].Biol Pharm Bull,2013,36(6):931-937.