發(fā)布時間：2018-03-14 01:05

  本文選題：腦缺血再灌注損傷　切入點：Bcl-2　出處：《青島大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的通過建立大鼠腦缺血再灌注損傷模型,觀察醒腦靜、丁苯酞及二者聯(lián)合應用分別對實驗性大鼠腦缺血再灌注損傷后神經細胞凋亡及Bcl-2和Bax表達的影響,明確醒腦靜、丁苯酞對大鼠腦缺血再灌注損傷后的腦保護作用。并通過Bcl-2和Bax表達的變化,探討其可能的作用機制。方法(1)60只成年雄性wistar大鼠(250±20g)隨機分為4組,模型組(n=15)、醒腦靜組(n=15)、丁苯酞組(n=15)、醒腦靜聯(lián)合丁苯酞組即聯(lián)合用藥組(n=15)。每組又分為6h、24h、72h三個亞組。(2)大鼠實驗性腦缺血再灌注模型(大腦中動脈閉塞(Middle cerebral artery occlusion,MCAO)模型)的制備采用改良線栓法。(3)細胞凋亡的檢測采用原位末端轉移酶標記技術(TUNEL);腦缺血再灌注損傷后腦組織形態(tài)學檢測采用HE染色,光鏡下觀察的方法;大鼠腦缺血再灌注各個時間點Bcl-2、Bax的表達的檢測采用免疫組化法。結果1.HE染色顯示:模型組造模側腦組織結構變得疏松,細胞間質水腫,部分形成空腔,著色變淺。神經細胞個數(shù)顯著減少,細胞胞體縮小,部分細胞輪廓不清,有不同程度的神經細胞變性、壞死。其中模型組破壞為著,醒腦靜組、丁苯酞組神經細胞破壞較輕,聯(lián)合用藥組神經細胞破壞最少。2.TUNEL染色顯示:TUNEL陽性細胞數(shù)總體趨勢是模型組最高,醒腦靜組及丁苯酞組不同程度降低,醒腦靜聯(lián)合丁苯酞組最低。丁苯酞組與模型組,聯(lián)合用藥組與模型組及丁苯酞組與聯(lián)合用藥組之間在6h、24h、72h時間點的差異均有統(tǒng)計學意義(P0.05),而醒腦靜組與模型組的比較僅24h時間點上差距有統(tǒng)計學意義(P0.05)。3.免疫組化顯示:丁苯酞組及聯(lián)合用藥組在6h、24h、72h時間點Bcl-2陽性表達較模型組均有提高。聯(lián)合用藥組Bcl-2陽性表達在各時間點均最高,醒腦靜組在各時間點與模型組比較差異無統(tǒng)計學意義(P0.05)。丁苯酞組及聯(lián)合用藥組在6h、24h、72h時間點Bax陽性表達較模型組均有降低(P0.05)。聯(lián)合用藥組Bax陽性表達在各時間點均最低,與丁苯酞組在各時間點比較差異有統(tǒng)計學意義(P0.05)。醒腦靜組在各時間點與模型組比較差異無統(tǒng)計學意義(P0.05)。Bcl-2于6h明顯開始升高,24h達高峰,同時Bax于6h表達較少,24h表達最低;醒腦靜聯(lián)合丁苯酞的聯(lián)合給藥組Bcl-2陽性表達在各時問點均最高。同時Bax陽性表達在各時間點均最低(P0.05)。結論1.醒腦靜、丁苯酞及二者聯(lián)合均可通過抑制腦缺血再灌注損傷后神經細胞凋亡來實現(xiàn)神經細胞保護作用,其中二者聯(lián)合效果最佳。2.丁苯酞可通過增加腦缺血再灌注損傷大鼠Bcl-2表達,減少Bax表達的方式來減少神經細胞凋亡,從而減輕腦缺血再灌注損傷。3.醒腦靜本身不能對Bcl-2、Bax的表達產生影響,但其可通過增強丁苯酞作用的方式影響B(tài)cl-2、Bax的表達,從而減輕腦缺血再灌注損傷。
[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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R743.3

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