本文關鍵詞：雙孔鉀通道TREK-1在急性腦缺血中的作用研究　出處：《蘭州大學》2014年碩士論文　論文類型：學位論文

  更多相關文章： REK-1 局灶性腦缺血 神經保護作用 血管擴張

【摘要】：目的：雙孔鉀通道TREK-1激活劑亞麻酸(ALA)干預急性腦缺血大鼠,觀察TREK-1通道激活對腦缺血的保護作用。 方法：線栓法制作大鼠急性腦缺血模型(MCAO),激光多普勒腦血流儀檢測缺血區(qū)域腦血流,將造模后大腦皮質局部腦血流下降≥30%的動物納入下一步實驗;實驗動物分為5組,分別為假手術組(N.S)、模型組(N.S)、亞麻酸小劑量組(ALA1uM),亞麻酸中劑量組(ALA10uM)和亞麻酸大劑量組(ALA100uM),10只/組；亞麻酸小、中、大劑量組動物經腦室分別給予1uM、10uM和100uM ALA,5uL/只,假手術組和模型組動物經腦室給予N.S5uL;激光多普勒腦血流儀再次檢測缺血區(qū)域腦血流;造模5小時后,單盲法對亞麻酸干預MCAO大鼠進行Garcia JH神經功能評分,激光多普勒再次記錄局部腦血流值；造模6小時后處死大鼠,斷頭取腦和基底動脈,實時熒光定量PCR和Western Blot法檢測TREK-1通道在腦皮質和基底動脈的表達情況。同時,腦組織冠狀位連續(xù)切片,紅四氮唑(TTC)染色計算腦梗死面積。 結果：1、與假手術組比較,模型組大鼠神經功能評分明顯下降,有統(tǒng)計學意義(P0.05)；亞麻酸各劑量組大鼠神經功能評分均增高,其中亞麻酸大、中劑量組動物神經功能評分升高與模型組比較具有統(tǒng)計學意義(P0.05)。與假手術組比較,模型組動物局部腦血流下降,有統(tǒng)計學意義(P0.01)：亞麻酸各劑量組大鼠局部腦血流均有不同程度升高,其中亞麻酸大、中劑量組大鼠腦血流升高與模型組比較具有統(tǒng)計學意義(P0.05)。模型組大鼠腦TTC染色腦梗死明顯；亞麻酸中、大劑量組大鼠腦TTC染色顯示腦梗死體積下降,其中亞麻酸大劑量組大鼠腦TTC染色腦梗死體積下降與模型組比較具有統(tǒng)計學意義(P0.05)。 2、Real-time PCR結果顯示,與假手術組比較,模型組與亞麻酸大劑量組大鼠腦皮質區(qū)TREK-1基因表達增加(P0.05)；與假手術組比較,各組大鼠基底動脈TREK-1基因表達無明顯差異(P0.05)。Western Blot結果顯示,假手術組、模型組和亞麻酸各劑量組大鼠腦皮質區(qū)均有TREK-1通道蛋白表達,其中以亞麻酸大劑量組TREK-1通道蛋白表達水平最高,其次是缺血組和假手術組。 結論：雙孔鉀通道TREK-1激活劑α-亞麻酸經腦室干預MCAO模型大鼠,可增加缺血區(qū)局部血流量,減輕腦組織缺血壞死,對腦缺血具有保護作用,與腦皮質TREK-1激活相關。
[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 偽 -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 偽 -linolenic acid through cerebral ventricle. Correlates with cortical TREK-1 activation.
【學位授予單位】：蘭州大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R743.3

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