瑞替加滨对缺血性脑卒中模型大鼠星形胶质细胞及Caspase-3的表达影响
发布时间：2018-07-25 08:54
【摘要】：目的通过Longa5方法比较神经功能缺损评分,TTC染色法测量脑梗死体积变化,应用免疫组化检测星形胶质纤维酸性蛋白(GFAP)阳性表达细胞(即星形胶质细胞)数和细胞凋亡蛋白酶-3(Caspase-3)阳性表达细胞数。探讨瑞替加滨对脑梗死大鼠GFAP、Caspase-3表达水平的影响,进而探讨瑞替加滨(Retigabine RTG)对缺血性脑卒中模型大鼠的保护作用及机制。方法选取SD大鼠75只,随机分为假手术组(15只,Sham组),模型组(15只,MCAO组),干预组(45只,RTG组)。干预组按照梗死再灌注后给药时间的不同又分为RTG 0h组;RTG 1h组;RTG 3h组,各组15只。通过大鼠尾静脉给药。给药剂量:RTG(10mg/kg)。假手术组和模型组不做任何处理,仅给予等量生理盐水。制备经典的大脑中动脉缺血再灌注模型大鼠,Longa5方法比较神经功能缺损评分,应用经典的TTC染色法测量脑梗死体积的变化;免疫组化法观察脑缺血后再灌注三组大鼠脑组织皮层GFAP、Caspase-3表达变化,显微镜下计数阳性细胞数。应用SPSS17.0统计软件进行统计学分析,结果用均数±标准(`X±s)表示,多组间比较采用单因素方差分析,两两比较采用LSD法或SNK法。P0.05认为差异有统计学意义。结果1神经功能缺损评分:假手术组未出现神经功能缺损症状,模型组组均表现出一定程度的神经功能缺损症状,表现为:或身体蜷缩,或身体平衡能力差,向左侧旋转及倾倒,或不能行走等。与模型组相比,RTG干预组神经功能缺损症状评分明显降低(P0.05),表现为单一的神经功能缺损症状,或者合并两种神经功能缺损症状,但RTG不同作用时间各亚组间差异无统计学意义。2 TTC染色:TTC图像显示:假手术组未见缺血梗死灶;模型组脑组织可见不同程度的梗死灶。RTG 0h、1 h、3 h干预组梗死灶较模型组明显降低(P0.05),但RTG不同作用时间各亚组间差异无统计学意义。3免疫组化:免疫组化检测假手术组可见少量GFAP及细胞凋亡Caspase-3阳性细胞的表达;干预组的GFAP、细胞凋亡Caspase-3表达均较模型组明显减少(P0.05),但干预组不同作用时间各亚组间差异无统计学意义。结论1瑞替加滨能明显降低脑梗死大鼠神经功能学评分,减少脑梗死体积。2瑞替加滨减少大鼠脑梗死中GFAP、细胞凋亡Caspase-3的表达,对大鼠缺血性脑卒中具有保护作用,其作用机制可能是瑞替加滨能够在细胞去极化时持续使大量钾离子外流抑制神经元的兴奋性,使神经细胞的兴奋性明显降低,减少谷氨酸的释放,从而进一步减轻脑梗死周围区的炎症反应,也抑制神经元的细胞凋亡,但时间依赖性有待研究。
[Abstract]:Objective to compare the changes of cerebral infarction volume by Longa5. The number of (GFAP) positive cells (astrocytes) and the number of apoptotic protease-3 (Caspase-3) positive cells were detected by immunohistochemistry. Objective: to investigate the effect of retegabine on the expression of Caspase-3 in cerebral infarction rats, and to explore the protective effect and mechanism of retegabine (Retigabine RTG) on ischemic stroke rats. Methods Seventy-five SD rats were randomly divided into sham-operated group (15 rats), model group (15 rats) and intervention group (45 rats). According to the time of administration after reperfusion, the intervention group was divided into RTG 0 h group and 1 h group with 15 rats in each group. The drug was administered through the tail vein of rats. Give the potion a dose of: RTG (10mg/kg). Sham operation group and model group did not do any treatment, only the same amount of physiological saline. A classical middle cerebral artery ischemia-reperfusion model was established to compare the neurological deficit scores in rats. The changes of cerebral infarct volume were measured by classical TTC staining. Immunohistochemical method was used to observe the changes of Caspase-3 expression in cerebral cortex of rats after cerebral ischemia and reperfusion, and the number of positive cells was counted under microscope. SPSS17.0 statistical software was used to carry out statistical analysis. The results were expressed as mean 卤standard (`X 卤s). The results showed that the differences were statistically significant by using LSD method or SNK method. Results 1 the neurological deficit score: there was no neurological deficit in the sham-operation group, and the model group showed some neurological deficit symptoms, such as body curling up or poor body balance. Rotate and fall to the left, or be unable to walk, etc. Compared with the model group, the score of neurological deficit symptoms in RTG intervention group was significantly lower (P0.05), showing a single neurological deficit symptom, or combined with two neurological deficit symptoms. However, there was no significant difference among subgroups of RTG at different time of action. 2. 2 TTC staining showed that there was no ischemic infarct in sham-operation group. The infarct focus of the model group was significantly lower than that of the model group (P0.05), but there was no significant difference between the subgroups of RTG at different time of action. Immunohistochemistry was used to detect sham-operation. A small amount of GFAP and apoptotic Caspase-3 positive cells were found in group A; The expression of GFAPand apoptotic Caspase-3 in intervention group was significantly lower than that in model group (P0.05), but there was no significant difference among subgroups of intervention group at different time of action. Conclusion (1) Retigabine can significantly decrease the neurological function score, reduce the volume of cerebral infarction, reduce the expression of GFAPand apoptosis Caspase-3 in cerebral infarction rats, and have protective effect on ischemic stroke in rats. The mechanism may be that retigabine can continuously inhibit the excitability of neurons and decrease the release of glutamate by retigabine, which can inhibit the excitability of neurons and inhibit the neuronal excitability by a large amount of potassium ion efflux during the depolarization of the cells. Therefore, the inflammatory reaction around cerebral infarction was further alleviated and the apoptosis of neurons was inhibited, but the time dependence needed to be studied.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3;R-332
本文编号：2143295
[Abstract]:Objective to compare the changes of cerebral infarction volume by Longa5. The number of (GFAP) positive cells (astrocytes) and the number of apoptotic protease-3 (Caspase-3) positive cells were detected by immunohistochemistry. Objective: to investigate the effect of retegabine on the expression of Caspase-3 in cerebral infarction rats, and to explore the protective effect and mechanism of retegabine (Retigabine RTG) on ischemic stroke rats. Methods Seventy-five SD rats were randomly divided into sham-operated group (15 rats), model group (15 rats) and intervention group (45 rats). According to the time of administration after reperfusion, the intervention group was divided into RTG 0 h group and 1 h group with 15 rats in each group. The drug was administered through the tail vein of rats. Give the potion a dose of: RTG (10mg/kg). Sham operation group and model group did not do any treatment, only the same amount of physiological saline. A classical middle cerebral artery ischemia-reperfusion model was established to compare the neurological deficit scores in rats. The changes of cerebral infarct volume were measured by classical TTC staining. Immunohistochemical method was used to observe the changes of Caspase-3 expression in cerebral cortex of rats after cerebral ischemia and reperfusion, and the number of positive cells was counted under microscope. SPSS17.0 statistical software was used to carry out statistical analysis. The results were expressed as mean 卤standard (`X 卤s). The results showed that the differences were statistically significant by using LSD method or SNK method. Results 1 the neurological deficit score: there was no neurological deficit in the sham-operation group, and the model group showed some neurological deficit symptoms, such as body curling up or poor body balance. Rotate and fall to the left, or be unable to walk, etc. Compared with the model group, the score of neurological deficit symptoms in RTG intervention group was significantly lower (P0.05), showing a single neurological deficit symptom, or combined with two neurological deficit symptoms. However, there was no significant difference among subgroups of RTG at different time of action. 2. 2 TTC staining showed that there was no ischemic infarct in sham-operation group. The infarct focus of the model group was significantly lower than that of the model group (P0.05), but there was no significant difference between the subgroups of RTG at different time of action. Immunohistochemistry was used to detect sham-operation. A small amount of GFAP and apoptotic Caspase-3 positive cells were found in group A; The expression of GFAPand apoptotic Caspase-3 in intervention group was significantly lower than that in model group (P0.05), but there was no significant difference among subgroups of intervention group at different time of action. Conclusion (1) Retigabine can significantly decrease the neurological function score, reduce the volume of cerebral infarction, reduce the expression of GFAPand apoptosis Caspase-3 in cerebral infarction rats, and have protective effect on ischemic stroke in rats. The mechanism may be that retigabine can continuously inhibit the excitability of neurons and decrease the release of glutamate by retigabine, which can inhibit the excitability of neurons and inhibit the neuronal excitability by a large amount of potassium ion efflux during the depolarization of the cells. Therefore, the inflammatory reaction around cerebral infarction was further alleviated and the apoptosis of neurons was inhibited, but the time dependence needed to be studied.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3;R-332
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