双孔钾通道TREK-1在急性脑缺血中的作用研究

发布时间：2018-01-08 05:14

本文关键词：双孔钾通道TREK-1在急性脑缺血中的作用研究　出处：《兰州大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：双孔钾通道TREK-1激活剂亚麻酸(ALA)干预急性脑缺血大鼠,观察TREK-1通道激活对脑缺血的保护作用。方法：线栓法制作大鼠急性脑缺血模型(MCAO),激光多普勒脑血流仪检测缺血区域脑血流,将造模后大脑皮质局部脑血流下降≥30%的动物纳入下一步实验;实验动物分为5组,分别为假手术组(N.S)、模型组(N.S)、亚麻酸小剂量组(ALA1uM),亚麻酸中剂量组(ALA10uM)和亚麻酸大剂量组(ALA100uM),10只/组；亚麻酸小、中、大剂量组动物经脑室分别给予1uM、10uM和100uM ALA,5uL/只,假手术组和模型组动物经脑室给予N.S5uL;激光多普勒脑血流仪再次检测缺血区域脑血流;造模5小时后,单盲法对亚麻酸干预MCAO大鼠进行Garcia JH神经功能评分,激光多普勒再次记录局部脑血流值；造模6小时后处死大鼠,断头取脑和基底动脉,实时荧光定量PCR和Western Blot法检测TREK-1通道在脑皮质和基底动脉的表达情况。同时,脑组织冠状位连续切片,红四氮唑(TTC)染色计算脑梗死面积。结果：1、与假手术组比较,模型组大鼠神经功能评分明显下降,有统计学意义(P0.05)；亚麻酸各剂量组大鼠神经功能评分均增高,其中亚麻酸大、中剂量组动物神经功能评分升高与模型组比较具有统计学意义(P0.05)。与假手术组比较,模型组动物局部脑血流下降,有统计学意义(P0.01)：亚麻酸各剂量组大鼠局部脑血流均有不同程度升高,其中亚麻酸大、中剂量组大鼠脑血流升高与模型组比较具有统计学意义(P0.05)。模型组大鼠脑TTC染色脑梗死明显；亚麻酸中、大剂量组大鼠脑TTC染色显示脑梗死体积下降,其中亚麻酸大剂量组大鼠脑TTC染色脑梗死体积下降与模型组比较具有统计学意义(P0.05)。 2、Real-time PCR结果显示,与假手术组比较,模型组与亚麻酸大剂量组大鼠脑皮质区TREK-1基因表达增加(P0.05)；与假手术组比较,各组大鼠基底动脉TREK-1基因表达无明显差异(P0.05)。Western Blot结果显示,假手术组、模型组和亚麻酸各剂量组大鼠脑皮质区均有TREK-1通道蛋白表达,其中以亚麻酸大剂量组TREK-1通道蛋白表达水平最高,其次是缺血组和假手术组。结论：双孔钾通道TREK-1激活剂α-亚麻酸经脑室干预MCAO模型大鼠,可增加缺血区局部血流量,减轻脑组织缺血坏死,对脑缺血具有保护作用,与脑皮质TREK-1激活相关。
[Abstract]:Aim: to investigate the protective effect of TREK-1 activator of double pore potassium channel (TREK-1) on cerebral ischemia in rats with acute cerebral ischemia. Methods: the rat model of acute cerebral ischemia was made by thread embolization, and the cerebral blood flow in ischemic area was measured by laser Doppler cerebral blood flow meter. The animals whose regional cerebral blood flow decreased more than 30% after the model were included in the next experiment. The experimental animals were divided into 5 groups: sham operation group, model group, small dose linolenic acid group. The middle dose of linolenic acid group (ALA10uM) and the high dose group of linolenic acid (ALA100uMN) (10 / group); Small, medium and large doses of linolenic acid were treated with 1uM10uM and 100uM ALA5uL respectively, and rats in sham-operated group and model group were given N.S5uL via ventricle. The cerebral blood flow in ischemic area was detected again by laser Doppler cerebral blood flow analyzer. After 5 hours of modeling, single blind method was used to evaluate the Garcia JH neurological function of MCAO rats with linolenic acid, and the regional cerebral blood flow was recorded again by laser Doppler. The rats were killed 6 hours later, and the brain and basilar artery were taken out. Real-time quantitative PCR and Western Blot were used to detect the expression of TREK-1 channel in cerebral cortex and basilar artery. The area of cerebral infarction was calculated by red tetrazolium TTC staining. Results compared with the sham operation group, the neurological function score of the model group decreased significantly (P 0.05). The scores of nerve function of rats in each dose group of linolenic acid were all increased, and the scores of nerve function in middle dose group were higher than those in the model group, which were significantly higher than those in the sham operation group (P 0.05). The regional cerebral blood flow in the model group was decreased with statistical significance (P 0.01): the regional cerebral blood flow in each dose group of linolenic acid was increased in varying degrees, among which linolenic acid was high. The increase of cerebral blood flow in the middle dose group was significantly higher than that in the model group (P 0.05). The cerebral infarction was obviously stained by TTC in the model group. In linolenic acid group, TTC staining showed that the volume of cerebral infarction decreased. The cerebral infarct volume in the large dose linolenic acid group was significantly lower than that in the model group by TTC staining (P 0.05). 2the results of Real-time PCR showed that compared with sham operation group, the expression of TREK-1 gene in cerebral cortex of model group and high dose group of linolenic acid increased (P 0.05). Compared with sham-operated group, the expression of TREK-1 gene in basilar artery of rats in each group was not significantly different. Western Blot showed that there was no significant difference in the expression of TREK-1 gene in basilar artery of rats in the sham-operation group. The expression of TREK-1 channel protein was found in the cortex of rats in the model group and linolenic acid groups, and the highest expression level of TREK-1 channel protein was found in the large dose linolenic acid group. The second was ischemia group and sham operation group. Conclusion: double hole potassium channel TREK-1 activator 伪 -linolenic acid can increase the regional blood flow in ischemic area, reduce ischemic necrosis of brain tissue and protect cerebral ischemia in MCAO model rats after intervention with 伪 -linolenic acid through cerebral ventricle. Correlates with cortical TREK-1 activation.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.3

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