缺血预适应对局灶性脑缺血再灌注大鼠低氧诱导因子-1α及存活素表达的影响

发布时间：2018-03-27 12:51

本文选题：缺血预适应　切入点：脑缺血再灌注　出处：《河北联合大学》2014年硕士论文

【摘要】：目的观察缺血预适应对局灶性脑缺血再灌注大鼠缺血灶周围皮质区及缺血侧海马CA1区低氧诱导因子-1α（HIF-1α）及存活素（Survivin）表达的影响，，探讨缺血预适应的脑保护机制。方法130只雄性SD大鼠，随机分为假手术组（SO组，n=10）、脑缺血组（MCAO组，n=60）、脑缺血预适应组（BIP组，n=60），后两组再分为缺血2h再灌注2h、6h、12h、24h、48h、72h6个观察时间点（每个时间点各10只，其中5只用于免疫组化染色，另外5只用于Western blot检测）。采用大脑中动脉两次线栓法制备大鼠脑缺血预适应模型。各组作如下处理：BIP组：大脑中动脉缺血10min为预适应条件，24h后行大脑中动脉缺血2h；MCAO组：以假手术代替预适应，余步骤同BIP组；SO组：以两次假手术代替预适应及再缺血。免疫组织化学染色及Western blot法检测缺血灶周围皮质区及缺血侧海马CA1区HIF-1α、Survivin的表达情况。结果1SO组皮质区HIF-1α阳性细胞表达水平低（4.77±1.11），蛋白表达量低（0.45±0.03）。MCAO组缺血灶周围皮质区HIF-1α阳性细胞、缺血侧皮质区HIF-1α蛋白量表达均于脑缺血再灌注2h升高，24h达各时间点最高水平，随后表达逐渐降低，72h时仍明显高于SO组。BIP组缺血灶周围皮质区HIF-1α阳性细胞及缺血侧皮质区HIF-1α蛋白量表达趋势与MCAO组一致，相同时间点HIF-1α阳性细胞数量及蛋白表达量均高于MCAO组，差异有统计学意义（P0.05）。MCAO组、BIP组HIF-1α阳性细胞数量及蛋白表达量均高于SO组，差异有统计学意义（P0.05）。2SO组海马CA1区HIF-1α阳性细胞的表达较低（4.80±1.30），蛋白表达量较低（0.35±0.03）。MCAO组缺血侧海马CA1区HIF-1α阳性细胞及蛋白量表达于脑缺血再灌注后2h升高，24h在各时间点表达水平最高，而后逐渐下降，72h时表达接近6h水平。BIP组缺血侧海马CA1区HIF-1α阳性细胞及蛋白量表达规律与MCAO组一致，除再灌注2h、6h外，相同时间点BIP组HIF-1α阳性细胞的数量高于MCAO组，差异有统计学意义（P0.05）；BIP组各时间点HIF-1α蛋白表达量均高于MCAO组，差异有统计学意义（P0.05）。MCAO组、BIP组HIF-1α阳性细胞数量及蛋白表达量均高于SO组，差异有统计学意义（P0.05）。3SO组皮质区可见少量Survivin阳性细胞表达（4.96±0.63），少量Survivin蛋白表达（0.59±0.03）。MCAO组于脑缺血再灌注2h缺血灶周围皮质区Survivin阳性细胞、缺血侧皮质区Survivin蛋白量表达增加，48h达各时间点最高水平，72h时表达接近24h水平。BIP组缺血灶周围皮质区Survivin阳性细胞、缺血侧皮质区Survivin蛋白量的表达趋势与MCAO组具备相同的时间规律性，除再灌注2h外，同一时间点BIP组Survivin阳性细胞表达数量高于MCAO组，差异有统计学意义（P0.05）；BIP组各时间点Survivin蛋白表达量较MCAO组高，两组比较差异有统计学意义（P0.05）。MCAO组与BIP组Survivin阳性细胞的表达数量及蛋白表达量均高于SO组，差异有统计学意义（P0.05）。4SO组海马CA1区Survivin阳性细胞表达量少（3.18±0.46），蛋白表达量少（0.39±0.03）。MCAO组于脑缺血再灌注2h缺血侧海马CA1区Survivin阳性细胞及蛋白量表达增加，24h在各时间点水平为最高，随后表达逐渐下降，72h时表达接近6h水平。BIP组缺血侧海马CA1区Survivin阳性细胞及蛋白量的表达趋势与MCAO组时间性一致，且各时间点Survivin阳性细胞的表达数量及蛋白表达量均高于MCAO组，差异有统计学意义（P0.05）。以上两组Survivin阳性细胞及蛋白的表达水平均高于SO组，差异有统计学意义（P0.05）。结论1缺血预适应可以促进局灶性脑缺血再灌注大鼠缺血灶周围皮质区及缺血侧海马CA1区HIF-1α、Survivin的表达。2缺血预适应可能通过HIF-1α、Survivin的表达上调参与脑缺血耐受的发生。
[Abstract]:Objective To investigate the effects of ischemic preconditioning on the expression of hypoxia - inducible factor - 1伪 ( HIF - 1伪 ) and survivin ( Survivin ) in the peripheral cortex and hippocampus CA1 region of ischemia - reperfusion rats and to explore the mechanism of ischemic preconditioning .

Methods 130 male SD rats were randomly divided into sham operation group ( SO group , n = 10 ) , cerebral ischemia group ( middle cerebral ischemia group , n = 60 ) and cerebral ischemia preconditioning group ( n = 60 ) . Two groups were divided into two groups : ischemia 2h reperfusion 2h , 6h , 12h , 24h , 48h , 72h6 observation time points ( 10 at each time point , 5 only for immunohistochemical staining and 5 for Western blot detection ) . The model of cerebral ischemia preconditioning was prepared by using two - thread method of middle cerebral artery occlusion . The following treatments were used : the cerebral artery ischemia was 10 min as the preconditioning condition , and the middle cerebral artery was ischemia for 2h after 24h ;
In the middle group , the sham operation was used to replace the pre - adaptation , and the remaining steps were the same as that of the bip group .
SO group : The expression of HIF - 1伪 and Survivin was detected by two false surgeries instead of preconditioning and ischemia . Immunohistochemical staining and Western blot were used to detect the expression of HIF - 1伪 and Survivin in the cortex and hippocampus CA1 region .

Results The expression of HIF - 1伪 positive cells and the expression of HIF - 1伪 in the cortex were higher than those in the middle cerebral ischemia - reperfusion group ( P0.05 ) .
There was a significant difference in the expression of Survivin positive cells ( 4.96 卤 0.63 ) and Survivin protein expression ( P 0.05 ) . The expression of Survivin positive cells in the cortical area around the ischemic foci was significantly higher than that in the middle cerebral ischemia group ( P 0.05 ) .
The expression of survivin - positive cells and protein in the CA1 region of the hippocampus CA1 region were significantly higher than those in the middle cerebral ischemia - reperfusion 2h group ( P0.05 ) .

Conclusion 1 Ischemia preconditioning can promote the expression of HIF - 1伪 and Survivin in the peripheral cortex and hippocampus CA1 region of focal cerebral ischemia - reperfusion rats . The expression of HIF - 1伪 and Survivin may be increased by the expression of HIF - 1伪 and Survivin in the ischemic preconditioning .

【学位授予单位】：河北联合大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.3

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