醒脑静联合丁苯肽对脑缺血再灌注损伤的脑保护及细胞凋亡的影响

发布时间：2018-03-14 01:05

本文选题：脑缺血再灌注损伤　切入点：Bcl-2　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的通过建立大鼠脑缺血再灌注损伤模型,观察醒脑静、丁苯酞及二者联合应用分别对实验性大鼠脑缺血再灌注损伤后神经细胞凋亡及Bcl-2和Bax表达的影响,明确醒脑静、丁苯酞对大鼠脑缺血再灌注损伤后的脑保护作用。并通过Bcl-2和Bax表达的变化,探讨其可能的作用机制。方法(1)60只成年雄性wistar大鼠(250±20g)随机分为4组,模型组(n=15)、醒脑静组(n=15)、丁苯酞组(n=15)、醒脑静联合丁苯酞组即联合用药组(n=15)。每组又分为6h、24h、72h三个亚组。(2)大鼠实验性脑缺血再灌注模型(大脑中动脉闭塞(Middle cerebral artery occlusion,MCAO)模型)的制备采用改良线栓法。(3)细胞凋亡的检测采用原位末端转移酶标记技术(TUNEL);脑缺血再灌注损伤后脑组织形态学检测采用HE染色,光镜下观察的方法;大鼠脑缺血再灌注各个时间点Bcl-2、Bax的表达的检测采用免疫组化法。结果1.HE染色显示:模型组造模侧脑组织结构变得疏松,细胞间质水肿,部分形成空腔,着色变浅。神经细胞个数显著减少,细胞胞体缩小,部分细胞轮廓不清,有不同程度的神经细胞变性、坏死。其中模型组破坏为著,醒脑静组、丁苯酞组神经细胞破坏较轻,联合用药组神经细胞破坏最少。2.TUNEL染色显示:TUNEL阳性细胞数总体趋势是模型组最高,醒脑静组及丁苯酞组不同程度降低,醒脑静联合丁苯酞组最低。丁苯酞组与模型组,联合用药组与模型组及丁苯酞组与联合用药组之间在6h、24h、72h时间点的差异均有统计学意义(P0.05),而醒脑静组与模型组的比较仅24h时间点上差距有统计学意义(P0.05)。3.免疫组化显示:丁苯酞组及联合用药组在6h、24h、72h时间点Bcl-2阳性表达较模型组均有提高。联合用药组Bcl-2阳性表达在各时间点均最高,醒脑静组在各时间点与模型组比较差异无统计学意义(P0.05)。丁苯酞组及联合用药组在6h、24h、72h时间点Bax阳性表达较模型组均有降低(P0.05)。联合用药组Bax阳性表达在各时间点均最低,与丁苯酞组在各时间点比较差异有统计学意义(P0.05)。醒脑静组在各时间点与模型组比较差异无统计学意义(P0.05)。Bcl-2于6h明显开始升高,24h达高峰,同时Bax于6h表达较少,24h表达最低;醒脑静联合丁苯酞的联合给药组Bcl-2阳性表达在各时问点均最高。同时Bax阳性表达在各时间点均最低(P0.05)。结论1.醒脑静、丁苯酞及二者联合均可通过抑制脑缺血再灌注损伤后神经细胞凋亡来实现神经细胞保护作用,其中二者联合效果最佳。2.丁苯酞可通过增加脑缺血再灌注损伤大鼠Bcl-2表达,减少Bax表达的方式来减少神经细胞凋亡,从而减轻脑缺血再灌注损伤。3.醒脑静本身不能对Bcl-2、Bax的表达产生影响,但其可通过增强丁苯酞作用的方式影响Bcl-2、Bax的表达,从而减轻脑缺血再灌注损伤。
[Abstract]:Objective to observe the effects of Xingnaojing, butyphthalide and their combination on neuronal apoptosis and the expression of Bcl-2 and Bax after cerebral ischemia-reperfusion injury in rats, and to observe the effects of Xingnaojing, butyphthalide and their combination on neuronal apoptosis and expression of Bcl-2 and Bax in rats with cerebral ischemia-reperfusion injury. The protective effect of butyphthalide on brain after cerebral ischemia-reperfusion injury in rats was studied. The possible mechanism of butyphthalide was investigated by the changes of Bcl-2 and Bax expressions. Methods 60 adult male wistar rats were randomly divided into 4 groups. Model group, Xingnaojing group, butyrophthalide group and butyrophthalide group respectively. Each group was divided into three subgroups (6 h, 24 h, 72 h), and the experimental cerebral ischemia reperfusion model (middle cerebral artery occlusion MCAO) was established in each group. In situ terminal transferase labeling technique was used to detect apoptosis, and cerebral histomorphology was detected by HE staining after cerebral ischemia-reperfusion injury. Immunohistochemical method was used to detect the expression of Bcl-2P Bax at different time points after cerebral ischemia-reperfusion in rats. Results 1. He staining showed that the brain structure of the model group became loose, the interstitial edema, and some cavities were formed. The number of nerve cells decreased significantly, the cell body shrank, the outline of some cells was unclear, and there were varying degrees of degeneration and necrosis of nerve cells, among which the model group was damaged, the Xingnaojing group and butyphthalide group were less damaged. The neuronal damage in combination group was the least. 2. Tunel staining showed that the total number of positive cells in the model group was the highest, Xingnaojing group and butyphthalide group were lower, Xingnaojing combined with butyphthalide group was the lowest, butyrophthalide group and model group were the lowest. The differences between the combined group and the model group and the butyphthalide group and the combined treatment group were statistically significant at the time points of 6h and 24h and 72h, but the difference between the Xingnaojing group and the model group was only significant at 24 hours. The results showed that the positive expression of Bcl-2 in butyphthalide group and combined treatment group was higher than that in model group at 6h and 24h / 72h, and the positive expression of Bcl-2 was the highest in combination group at each time point. There was no significant difference between Xingnaojing group and model group at each time point (P 0.05). The positive expression of Bax in butadiphthalide group and combined treatment group was significantly lower than that in model group at 24 h and 72 h. The positive expression of Bax in combination group was the lowest at all time points. There was no significant difference between Xingnaojing group and model group at each time point (P 0.05). The expression of Bax began to increase at 6 h and reached the peak at 24 h, while the expression of Bax was lower at 6 h than that in the model group (P 0. 05%, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05). The positive expression of Bcl-2 was the highest at each time point, and the expression of Bax was the lowest in each time point. Conclusion 1.Xingnaojing, xingnaojing, and butyphthalide combined with butyphthalide have the highest positive expression of Bcl-2 at each time point. The protective effect of butyphthalide and its combination can be achieved by inhibiting neuronal apoptosis after cerebral ischemia-reperfusion injury, and the best combination of them is .2.Butylphthalide can increase the expression of Bcl-2 in rats with cerebral ischemia-reperfusion injury. Xingnaojing itself can not affect the expression of Bcl-2P Bax, but it can affect the expression of Bcl-2O Bax by enhancing the effect of butyphthalide. So as to reduce cerebral ischemia reperfusion injury.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3

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