本文選題：Bcl-2 + Bax��； 參考：《華中科技大學(xué)》2006年碩士論文

【摘要】： Bcl-2、BaxCaspase-3是與凋亡相關(guān)的三種蛋白。其中Bax和Bcl-2是細(xì)胞凋亡通路中的一對(duì)促凋亡和抗凋亡的相互影響的相關(guān)蛋白,二者之間的比例決定了細(xì)胞的命運(yùn)是生存還是凋亡,Caspase-3是凋亡過(guò)程中最重要的蛋白酶,是多種凋亡途徑的共同下游效應(yīng)部分,是細(xì)胞凋亡蛋白酶級(jí)聯(lián)反應(yīng)的必經(jīng)之路。本研究探討了疼痛刺激對(duì)大鼠脊髓背角神經(jīng)元凋亡相關(guān)蛋白表達(dá)的影響及年齡相關(guān)性變化。對(duì)各組大鼠進(jìn)行行為學(xué)測(cè)試,記錄各組大鼠在疼痛刺激1h內(nèi)每5min的縮腿舔爪時(shí)間,觀察不同年齡大鼠對(duì)疼痛刺激的反應(yīng);應(yīng)用免疫組織化學(xué)方法對(duì)Caspase-3在各組大鼠脊髓背角的表達(dá)進(jìn)行分析并比較刺激側(cè)與未刺激側(cè)的差別,用western-blotting技術(shù)檢測(cè)Caspase-3,Bcl-2,Bax在各組大鼠脊髓背角神經(jīng)元的表達(dá)并計(jì)算Bax與Bcl-2的比值。結(jié)果1、行為學(xué)測(cè)試顯示在同等刺激的條件下老中青三組大鼠的縮腿舔爪時(shí)間差異顯著(P 0.05),其中青年組大鼠縮腿舔爪時(shí)間最長(zhǎng),中年組大鼠其次,老年組大鼠最短。2、免疫組織化學(xué)顯示:在未刺激側(cè)和刺激側(cè),老中青三組表達(dá)Caspase-3免疫反應(yīng)陽(yáng)性產(chǎn)物光密度值均差別明顯(P0.05),均為老年組光密度值最高,中年組次之,青年組最低;老中、青各年齡組大鼠疼痛刺激側(cè)Caspase-3的表達(dá)均較未刺激側(cè)明顯增加(P 0.05);其中青年組增幅明顯高于中老年組(P 0.05),中老年組之間無(wú)顯著性差異(P 0.05)。3、Western-blotting顯示:各組Caspase-3的表達(dá)與免疫組織化學(xué)結(jié)果基本一致。而B(niǎo)ax/Bcl-2表達(dá)結(jié)果為:未刺激側(cè),老年組和中年組Bax/Bcl-2均較青年組高(P0.05),中、老年組之間無(wú)顯著性差異(P0.05);刺激側(cè),也是老年組和中年組Bax/Bcl-2均較青年組高(P0.05),老年組與中年組之間無(wú)顯著性差異(P0.05);各組大鼠刺激側(cè)相對(duì)于未刺激側(cè)Bax/ Bcl-2明顯增加(P0.05),其中青年組增幅明顯高于中老年組(P0.05),中老年組之間無(wú)顯著性差異(P0.05)。 結(jié)論(1)在未刺激側(cè)和刺激側(cè),凋亡相關(guān)蛋白(Caspase-3,Bax/Bcl-2)的表達(dá)均有明顯的年齡性變化,即老年組和中年組表達(dá)較強(qiáng),青年組較弱; (2)疼痛刺激可以誘導(dǎo)脊髓背角神經(jīng)元凋亡; (3)疼痛刺激對(duì)青年大鼠脊髓背角神經(jīng)元凋亡的影響最大;
[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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R363

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