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  本文選題：意向性運動　切入點：局灶性腦缺血　出處：《中南大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的：通過觀察意向性運動對局灶性腦缺血大鼠神經功能及缺血半暗帶ERK-CREB通路表達及GluR2與PICK1共定位表達比率的影響,初步探討意向性運動對局灶性腦缺血后神經功能的影響及其可能的作用機制。 方法：雄性Sprague-Dawley (SD)大鼠144只,隨機均分為大腦中動脈栓塞模型(MCAO)組、游泳運動(SE)組、飼養(yǎng)環(huán)境改變(EM)組和意向性運動(WM)組,各組又分7d、15d、30d三個亞組。采用線栓法制備MCAO大鼠模型,動態(tài)觀察各組大鼠行為學變化,TTC染色觀察不同運動方式對15d組大鼠腦梗死體積的影響,RT-PCR及免疫熒光單標染色動態(tài)檢測缺血半暗帶腦組織ERK、CREB、GluR2mRNA及pERK、pCREB蛋白的表達水平,免疫熒光雙標染色檢測15d組GluR2與PICK1的共定位表達比率。 結果： 1.各組大鼠在腦缺血再灌注2h時的神經功能評分最高,1d后略下降,第3d略有回升,以后隨時間推移各組評分均呈下降趨勢,各組評分差別無統(tǒng)計學意義(P0.05)。SE組評分在再灌7d較MCAO組升高(P0.05),而再灌15、30d下降(P0.05)；EM組及WM組評分在再灌7、15、30d,均較MCAO組下降(P0.05),以WM組下降更明顯。EM組及WM組大鼠的爬梯頻率均呈逐漸增加趨勢,且WM組較EM組顯著增加(P0.05)。 2.TTC染色結果顯示：腦缺血再灌注15d時,與MCAO組比較,SE組梗死體積差異無統(tǒng)計學意義(P0.05)；EM組及WM組腦梗死體積減小(P0.05),以WM組更明顯(P0.05)。 3. RT-PCR結果顯示：與MCAO組比較,SE組腦缺血再灌注7d時ERK、CREB mRNA的表達減少(P0.05),15、30d后的表達明顯增加(P0.05),而GluR2mRNA的表達在7d、15d、30d時均明顯增加(P0.05)；EM組及WM組各實驗點ERK、CREB、GluR2mRNA的表達較MCAO組均增加(P0.05),以WM組更明顯(P0.05)。 4.免疫熒光單標染色結果顯示：pERK在胞核、胞漿、胞膜均有表達,pCREB主要表達于胞核。MCAO組在腦缺血再灌注15d時,pERK.pCREB表達較7d時上調,隨后基本保持恒定；SE組、EM組、WM組在再灌注7、15、30d,兩蛋白的表達隨時間呈上升趨勢；但SE組與MCAO組比較,兩蛋白表達在再灌7d時減少(P0.05),15、30d明顯增加(P0.05)；EM組在各時間點兩蛋白的表達均較MCAO組增多(P0.05)；WM組兩蛋白的表達在各時間點均較其他三組明顯增多(P0.05)。 5.免疫熒光雙標結果顯示：GluR2與PICK1蛋白均表達于胞漿和胞膜,且兩者的分布情況及變化趨勢存在高度一致性。腦缺血再灌注15d,SE組、EM組及WM組GluR2、PICK1的陽性細胞數(shù)均較MCAO組明顯增加,而WM組雙標陽性細胞數(shù)較其他三組明顯增加(P0.05) 結論： 意向性運動能改善局灶性腦缺血大鼠的神經功能,減小腦梗死體積,且較游泳運動有更明顯的優(yōu)勢,可能與其上調并激活缺血半暗帶腦組織ERK-CREB通路,并增強GluR2與PICK1的相互作用有關。
[Abstract]:Objective: to observe the effects of intentionality exercise on the expression of ERK-CREB pathway in ischemic penumbra and the ratio of co-localization of GluR2 and PICK1 in focal cerebral ischemia rats. To explore the effect of intentionality exercise on neural function after focal cerebral ischemia and its possible mechanism. Methods: a total of 144 male Sprague-Dawley SD rats were randomly divided into three groups: middle cerebral artery embolization (MCAO) group, swimming exercise group (Sprague-Dawley) group, feeding environment change group and intentionality exercise group. Each group was divided into three subgroups: 7 d, 15 d and 30 d, respectively. The effects of different exercise modes on cerebral infarct volume in rats of 15 d group were observed dynamically by TTC staining. RT-PCR and immunofluorescence staining were used to dynamically detect the expression of ERKN CREBN GluR2 mRNA and pCREB protein in ischemic penumbra. The co-localization ratio of GluR2 and PICK1 in 15 d group was detected by double immunofluorescence staining. Results:. 1. After 2 hours of cerebral ischemia and reperfusion, the neurological function scores of the rats in each group decreased slightly after 1 day, then increased slightly on the third day, and then decreased with the passage of time. There was no significant difference in the scores of each group. The scores of group E were significantly higher than those of group MCAO on the 7th day after reperfusion, while the scores of group EM and group WM were lower than those of group MCAO on the 7th day after reperfusion. The climbing frequency of rats in the WM group was more obvious than that of the rats in the group of WM and the group of EM and WM, and the scores of the EM group and the WM group were lower than those of the group of MCAO on the 7th day after reperfusion, and the scores of the EM group and the WM group were lower than those of the group of MCAO on the 7th day after reperfusion. All showed a trend of gradual increase, Compared with EM group, WM group significantly increased P0.05. 2. The results of TTC staining showed that at 15 days after cerebral ischemia and reperfusion, there was no significant difference in infarct volume between SE group and MCAO group. 3. RT-PCR results showed that compared with MCAO group, the expression of ERKO CREB mRNA in SE group decreased significantly after 15 days of cerebral ischemia and reperfusion compared with that in MCAO group, and the expression of GluR2mRNA increased significantly at the end of the 15th day after reperfusion, while the expression of GluR2mRNA increased significantly in EM group and WM group on the 7th day, 15 days after reperfusion and 30 days after reperfusion, and the expression of ERKB CREB GluR2 mRNA was significantly higher than that in MCAO group. The addition of P0.05 was more obvious in WM group. 4. The results of immunofluorescence single label staining showed that the expression of pCREB in nucleus, cytoplasm and cell membrane was mainly up-regulated at 15 days after cerebral ischemia and reperfusion in MCAO group, and the expression of pERK.pCREB was up-regulated at 7 days after cerebral ischemia and reperfusion. After 30 days of reperfusion, the expression of two proteins increased in SE group, EM group and MCAO group, but compared with MCAO group, the expression of two proteins in SE group was higher than that in MCAO group. The expression of two proteins decreased on the 7th day after reperfusion. The expression of two proteins in the P0.05A EM group was significantly higher than that in the MCAO group at each time point, and the expression of the two proteins in the P0.05G group was significantly higher than that in the other three groups at each time point compared with the other three groups. 5. The results of immunofluorescence double labeling showed that both the protein of GluR2 and PICK1 were expressed in cytoplasm and membrane, and the distribution and change trend of them were highly consistent. The number of GluR2PICK1 positive cells in EM group and WM group were significantly higher than that in MCAO group on the 15th day after cerebral ischemia reperfusion, and the expression of GluR2PICK1 protein in both groups was significantly higher than that in MCAO group. The number of double-labeled positive cells in WM group was significantly higher than that in the other three groups (P0.05). Conclusion:. Intention exercise can improve the neural function and reduce the volume of cerebral infarction in rats with focal cerebral ischemia. It may be related to the up-regulation and activation of ERK-CREB pathway in ischemic penumbra. The interaction between GluR2 and PICK1 was enhanced.
【學位授予單位】：中南大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R743.3

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