低氧预适应减轻大鼠缺血—再灌注脑损伤中凋亡及相关蛋白表达的变化

发布时间：2018-01-08 04:28

本文关键词：低氧预适应减轻大鼠缺血—再灌注脑损伤中凋亡及相关蛋白表达的变化　出处：《山西医科大学》2006年硕士论文　论文类型：学位论文

【摘要】： 目的:探讨低氧预适应减轻大鼠缺血-再灌注性脑损伤中,凋亡及相关蛋白和HSP70表达的变化。方法:采用SD大鼠局灶性脑缺血再灌注模型(transient middle cerebral artery occlusion,MCAO),将48只大鼠随机分为3组6个亚组:假手术组(Sham)、缺血再灌注组(ischmia/reperfusion I/R)、低氧预适应组(hypoxic preconditioning HP+I/R),应用免疫组织化学和细胞死亡原位末端标记(Insitu end labeling ,ISEL)法研究脑组织细胞凋亡及凋亡相关蛋白Survivin、fas/fas-L和应激蛋白HSP-70的表达情况;并在缺血(ischemia,I)和再灌注(reperfusion,R)I/R后不同时间点(I1h、I2h、I3h、R1h、R4h、R8h、R24h)对大鼠进行神经体征的观察,记录动物神经行为缺陷计分,通过Y-电迷宫测试缺血再灌注24h后大鼠学习、记忆能力。结果: (1)缺血3h再灌注24h后神经功能缺陷计分及Y-电迷宫测试大鼠学习记忆能力(N代表学会标准时的训练次数;A/10代表记忆保持率): Sham组: 0分,N为47.5±7.1 , A/10为0.91±0.08; I/R组: 1.1±0.6分,N为86.3±9.2, A/10为0.54±0.14; HP+I/R组: 0.4±0.5分,N为66.3±7.4, A/10为0.66±0.09。 (2)高倍视野(high power field ,HPF)下Survivin蛋白阳性细胞数: Sham组为0.6±0.9个/HPF,I/R组为14.0±2.1个/HPF, HP+I/R组为27.1±4.2个/HPF。 (3)高倍视野下fas蛋白阳性细胞数: Sham组为0.8±1.2个/HPF,I/R组为27.3±2.8个/HPF, HP+I/R组为11.1±2.2个/HPF。 (4)高倍视野下fas-L蛋白阳性细胞数: Sham组为0.5±0.8个/HPF ,I/R组为24.6±3.6个/HPF, HP+I/R组为9.6±1.7个/HPF。 (5)高倍视野下HSP-70蛋白阳性细胞数: Sham组为0.3±0.5个/ HPF,I/R组为26.3±3.6个/HPF, HP+I/R组为47.0±3.1个/HPF。 (6)高倍视野下凋亡细胞数: Sham组为0.5±0.7个/HPF,I/R组为39.4±2.4个/HPF, HP+I/R组为23.1±4.4个/HPF。高倍视野下Survivin、fas/fas-L和HSP-70蛋白阳性细胞数、凋亡细胞数比较,HP+I/R组与I/R组及Sham组间差异具有统计学意义(p0.05)。缺血3h再灌注24h后神经功能缺陷计分及Y-电迷宫测试大鼠学习记忆能力,HP+I/R组优于I/R组,差异有统计学意义(p0.05)。结论:低氧预适应可减轻局灶性缺血再灌注脑损伤,减少大鼠脑缺血再灌注后的脑细胞凋亡和神经功能缺失,提高动物学习记忆能力和损伤后神经功能恢复。上调神经细胞中Survivin、HSP-70蛋白表达;下调fas/fas-L蛋白表达可能是其发挥保护作用的分子机制之一。
[Abstract]:Aim: to investigate the changes of apoptosis and the expression of related proteins and HSP70 in hypoxic preconditioning rats. Methods: transient middle cerebral artery occlusion was used in SD rats with focal cerebral ischemia-reperfusion. MCAO, 48 rats were randomly divided into 3 groups and 6 subgroups: sham-operated group and ischemia-reperfusion group. Hypoxic preconditioning HP I / R in hypoxic preconditioning group. In situ end labeling was labeled by immunohistochemistry and in situ end labeling of cell death. The expression of apoptotic and apoptosis-related protein survivin fas-L and stress protein HSP-70 was studied by ISELL method. At different time points after I / R / R / R, I _ 1h ~ I _ 2h ~ I _ 3h ~ (1) ~ (1h) ~ (1) h ~ (4h) ~ (4) h ~ (8) h ~ (8 h) after ischemia / reperfusion / reperfusion / reperfusion / reperfusion / reperfusion / reperfusion / reperfusion / reperfusion / reperfusion / reperfusion / reperfusion. The neurological signs of rats were observed at 24 h, and the scores of neurobehavioral defects were recorded. The learning and memory abilities of rats after 24 hours of ischemia reperfusion were tested by Y- electrical maze. Results: 1) after 3 h of ischemia and 24 h after reperfusion, the scores of neural function defect and the training times of learning and memory ability in Y- electrical maze test were measured on behalf of the standard of learning. In Sham group, N was 47.5 卤7.1, A / 10 was 0.91 卤0.08; I / R group: 1.1 卤0.6 min N was 86.3 卤9.2, A / 10 was 0.54 卤0.14; HP I / R group: 0.4 卤0.5 min N 66.3 卤7.4, A / 10 0.66 卤0.09. (2) the number of Survivin protein positive cells in high power power field (Sham) group was 0.6 卤0.9 / HPF. The I / R group was 14.0 卤2.1 / HPFs, and the HP / I / R group was 27.1 卤4.2 / HPFs. The number of fas protein positive cells in Sham group was 0.8 卤1.2 / HPFN / I / R group (27.3 卤2.8 / HPF). The HP I / R group was 11.1 卤2.2 / HPF. (4) the number of fas-L protein positive cells in the Sham group was 0.5 卤0.8 / HPF-I / R group (24.6 卤3.6 / HPF). The HP I / R group was 9.6 卤1.7 / HPF. (5) the number of HSP-70 protein positive cells in Sham group was 0.3 卤0.5 / HPF-I / R group 26.3 卤3.6 / HPF. The HP I / R group was 47.0 卤3.1 / HPFs. (6) the number of apoptotic cells in the high power field: 0.5 卤0.7% HPF / R group was 39.4 卤2.4% HPF in Sham group. The HP I / R group was 23.1 卤4.4 / HPFs. The number of survivin fas-l and HSP-70 protein positive cells and the number of apoptotic cells were compared in high power field. The difference between HP / I / R group and I / R group and Sham group was statistically significant (p 0.05). After 3 hours of ischemia and 24 hours of reperfusion, the scores of nerve dysfunction and the learning and memory ability of rats were measured by Y- electrical maze. HP I / R group was better than I / R group, the difference was statistically significant (p 0.05). Conclusion: hypoxia preconditioning can reduce focal ischemia-reperfusion injury, reduce brain cell apoptosis and neural function loss after cerebral ischemia-reperfusion in rats. The expression of survivin HSP-70 protein in nerve cells was up-regulated by improving the ability of learning and memory and the recovery of nerve function after injury. Down-regulation of fas/fas-L protein expression may be one of its protective molecular mechanisms.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

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