疼痛对不同年龄大鼠脊髓背角Bcl-2、Bax及Caspase-3表达的影响

发布时间：2018-07-06 11:21

本文选题：Bcl-2 + Bax　；参考：《华中科技大学》2006年硕士论文

【摘要】： Bcl-2、BaxCaspase-3是与凋亡相关的三种蛋白。其中Bax和Bcl-2是细胞凋亡通路中的一对促凋亡和抗凋亡的相互影响的相关蛋白,二者之间的比例决定了细胞的命运是生存还是凋亡,Caspase-3是凋亡过程中最重要的蛋白酶,是多种凋亡途径的共同下游效应部分,是细胞凋亡蛋白酶级联反应的必经之路。本研究探讨了疼痛刺激对大鼠脊髓背角神经元凋亡相关蛋白表达的影响及年龄相关性变化。对各组大鼠进行行为学测试,记录各组大鼠在疼痛刺激1h内每5min的缩腿舔爪时间,观察不同年龄大鼠对疼痛刺激的反应;应用免疫组织化学方法对Caspase-3在各组大鼠脊髓背角的表达进行分析并比较刺激侧与未刺激侧的差别,用western-blotting技术检测Caspase-3,Bcl-2,Bax在各组大鼠脊髓背角神经元的表达并计算Bax与Bcl-2的比值。结果1、行为学测试显示在同等刺激的条件下老中青三组大鼠的缩腿舔爪时间差异显著(P 0.05),其中青年组大鼠缩腿舔爪时间最长,中年组大鼠其次,老年组大鼠最短。2、免疫组织化学显示:在未刺激侧和刺激侧,老中青三组表达Caspase-3免疫反应阳性产物光密度值均差别明显(P0.05),均为老年组光密度值最高,中年组次之,青年组最低;老中、青各年龄组大鼠疼痛刺激侧Caspase-3的表达均较未刺激侧明显增加(P 0.05);其中青年组增幅明显高于中老年组(P 0.05),中老年组之间无显著性差异(P 0.05)。3、Western-blotting显示:各组Caspase-3的表达与免疫组织化学结果基本一致。而Bax/Bcl-2表达结果为:未刺激侧,老年组和中年组Bax/Bcl-2均较青年组高(P0.05),中、老年组之间无显著性差异(P0.05);刺激侧,也是老年组和中年组Bax/Bcl-2均较青年组高(P0.05),老年组与中年组之间无显著性差异(P0.05);各组大鼠刺激侧相对于未刺激侧Bax/ Bcl-2明显增加(P0.05),其中青年组增幅明显高于中老年组(P0.05),中老年组之间无显著性差异(P0.05)。结论(1)在未刺激侧和刺激侧,凋亡相关蛋白(Caspase-3,Bax/Bcl-2)的表达均有明显的年龄性变化,即老年组和中年组表达较强,青年组较弱; (2)疼痛刺激可以诱导脊髓背角神经元凋亡; (3)疼痛刺激对青年大鼠脊髓背角神经元凋亡的影响最大;
[Abstract]:BCL-2 and BaxCaspase-3 are three kinds of proteins associated with apoptosis. Bax and Bcl-2 are a pair of apoptosis-promoting and anti-apoptotic related proteins in the apoptosis pathway. The ratio of Bax and Bcl-2 determines whether the fate of cells is survival or apoptosis Caspase-3 is the most important protease in the process of apoptosis. It is the common downstream effect of multiple apoptotic pathways and the only way to cascade the apoptotic protease. The purpose of this study was to investigate the effect of pain stimulation on the expression of apoptosis-related protein in spinal dorsal horn neurons and its age-related changes. Behavioral tests were performed on rats in each group, and the time of retraction and licking of paws per 5min was recorded during 1 hour of pain stimulation, and the response of rats of different ages to pain stimulation was observed. The expression of Caspase-3 in the spinal dorsal horn of rats in each group was analyzed by immunohistochemical method and the difference between the stimulated side and the unstimulated side was compared. The expression of Caspase-3 and Bcl-2Ba-Bax in the neurons of spinal dorsal horn in each group was detected by western-blotting technique and the ratio of Bax to Bcl-2 was calculated. Results 1Behavioral tests showed that they were aged under the same stimuli. Middle? There was significant difference between the three groups (P 0.05), in which the rats in the young group had the longest time of shrinking and licking the paw, the middle-aged rats were the second, the rats in the old group were the shortest. 2. Immunohistochemistry showed that in the unstimulated side and the stimulated side, the old rat had the longest leg and licking claw time (P 0.05). Middle? The optical density of Caspase-3 immunoreactive products in the three groups was significantly different (P0.05). The optical density of the aged group was the highest, the middle age group was the second, and the young group was the lowest. The expression of Caspase-3 in the pain stimulated side was significantly higher in the middle and young age groups than in the unstimulated side (P 0.05), and the increase in the young group was significantly higher than that in the middle and middle age group. Aged group (P 0.05), middle? There was no significant difference between the old group (P 0.05). 3 Western-blotting showed that the expression of Caspase-3 in each group was basically consistent with the immunohistochemical results. The results of Bax-P Bcl-2 expression were as follows: on the unstimulated side, the Bax-P Bcl-2 expression in the aged group and the middle-aged group was higher than that in the young group (P0.05), but there was no significant difference between the middle and old groups (P0.05), while in the middle and middle aged group, there was no significant difference (P0.05). The Bax-Bcl-2 in the aged group and the middle-aged group were higher than that in the young group (P0.05), there was no significant difference between the aged group and the middle-aged group (P0.05); the Bax-Bcl-2 ratio in the stimulated side of the rats was significantly higher than that in the non-stimulated side in each group (P0.05), and the increase in the young group was significantly higher than that in the middle-aged group. Elderly group (P0.05), middle? There was no significant difference between the old group (P0.05). Conclusion (1) the expression of Caspase-3 Bax-Bax / Bcl-2 in the unstimulated and stimulated sides has obvious age changes, that is, the expression of Caspase-3 Bax-Bcl-2 is stronger in the aged and middle-aged groups. (2) pain stimulation could induce apoptosis of spinal dorsal horn neurons; (3) pain stimulation could induce spinal dorsal horn neurons in young rats. The effect of apoptosis was the greatest.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

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