本文選題：自噬 + 缺血預處理��； 參考：《河北聯(lián)合大學》2014年碩士論文

【摘要】：目的研究自噬相關蛋白Beclin-1、LC3-II在缺血預處理后不同時間間隔大鼠腦缺血再灌注海馬中的表達，探討自噬在腦缺血預處理神經保護作用的可能機制。方法1.健康雄性SD大鼠50只隨機分為3組：假手術組、缺血再灌注組、缺血預處理組，缺血預處理組再按不同缺血預處理間隔時間分為24h、72h、5d3個亞組。2.參照Zea-Longa線栓法制作大鼠大腦中動脈缺血預處理和缺血再灌注模型。3.利用TTC染色、photoshop圖像分析法、HE染色、免疫組織化學染色、觀察腦梗死體積、海馬神經細胞病理變化、海馬神經細胞自噬相關蛋白Beclin-1及LC3-II表達的情況。4.透射電鏡觀察神經細胞中自噬超微結構的變化。5.比較各組神經功能缺失、腦梗死體積、海馬神經細胞病理變化、海馬神經細胞自噬相關蛋白Beclin-1及LC3-II表達、自噬體和各級溶酶體數(shù)量。6.以Excel數(shù)據(jù)庫整理后采用SPSS17.0統(tǒng)計學軟件進行數(shù)據(jù)分析，所得數(shù)據(jù)均以x±s表示，P0.05認為差異有統(tǒng)計學意義。采用Microsoft Excel軟件作圖。 結果1.缺血預處理組神經功能缺失癥狀較缺血再灌注組有所改善，差異具有統(tǒng)計學意義（P0.05）。2.與缺血再灌注組相比，缺血預處理組腦梗死體積縮小，差異具有統(tǒng)計學意義（P0.05）。3.HE染色可見假手術組神經細胞無損傷；與缺血再灌注組相比，缺血預處理各亞組海馬神經細胞損傷減輕。4.與假手術組相比，缺血再灌注組、缺血預處理組Beclin-1的表達均明顯增強，差異具有統(tǒng)計學意義（P0.05）；缺血預處理組與缺血再灌注組比較，可見Beclin-1陽性細胞數(shù)減少，差異具有統(tǒng)計學意義（P0.05）。5.假手術組只見極少量LC3-II陽性細胞表達，而在缺血再灌注組及缺血預處理組均可見大量LC3-II陽性細胞，差異有統(tǒng)計學意義（P0.05）。缺血預處理各亞組LC3-II陽性細胞表達顯著低于缺血再灌注組，差異有統(tǒng)計學意義（P0.05）。6.透射電鏡下缺血預處理組及缺血再灌注組均可見數(shù)量不等、處于不同期間的自噬體和自噬溶酶體，其數(shù)量在缺血預處理組明顯減少。7.缺血預處理72h亞組相比較24h亞組和5d亞組腦梗死體積縮小（P0.05），，海馬神經細胞損傷減輕，Beclin-1、LC3-II陽性細胞數(shù)不同程度減少（P0.05），自噬體的形成和溶酶體的激活減少。 結論1.采用改良后Zea-Longa線栓法制作的大鼠大腦中動脈缺血預處理和缺血再灌注模型方法簡便、安全、數(shù)據(jù)可靠。2.缺血預處理可減低缺血再灌注24小時后大鼠神經功能障礙評分、腦梗死體積、海馬神經細胞損傷，對缺血再灌注后神經細胞損傷具有保護作用。3.缺血預處理可能通過下調缺血再灌注24小時后自噬相關蛋白Beclin-1、LC3-II的表達，以減少缺血再灌注損傷神經細胞的自噬性死亡，減輕神經功能缺損癥狀，縮小腦梗死體積，從而發(fā)揮其腦保護作用。4.缺血預處理72h亞組腦梗死體積低于缺血預處理24h、5d亞組，Beclin-1、LC3-II陽性細胞數(shù)低于缺血預處理24h、5d亞組，偶見自噬體，各級溶酶體數(shù)量相對較少，提示最佳預處理誘導時間間隔為72h。
[Abstract]:Objective to study the expression of autophagy related protein Beclin-1 and LC3-II in hippocampus of ischemic reperfusion rats at different time intervals after ischemic preconditioning, and to explore the possible mechanism of autophagy in ischemic preconditioning. Method 1. 50 healthy male SD rats were randomly divided into 3 groups: sham operation group, ischemia reperfusion group, ischemic preconditioning group, The ischemic preconditioning group was then divided into 24h, 72h, 5d3 subgroup and 5d3 subgroup,.2. referred to Zea-Longa line embolus to make the rat middle cerebral artery ischemic preconditioning and ischemia reperfusion model.3. using TTC staining, Photoshop Image analysis, HE staining, immunohistochemical staining, observation of cerebral infarction volume and hippocampal neurocytopathic disease. Changes in the expression of autophagy related protein Beclin-1 and LC3-II in hippocampal neurons.4. transmission electron microscope observation of the ultrastructure changes of autophagy in the nerve cells.5. compared with the absence of nerve function, the volume of cerebral infarction, the pathological changes of hippocampal neurons, the expression of autophagic related protein Beclin-1 and LC3-II in the hippocampal neurons, autophago and each The number.6. of the grade lysosome was analyzed by the SPSS17.0 statistics software after the Excel database. The data were all expressed in X + s, and P0.05 thought the difference was statistically significant. The Microsoft Excel software was used to map.
Results there was a significant difference between the 1. ischemic preconditioning group and the ischemia reperfusion group. The difference was statistically significant (P0.05) compared with the ischemia reperfusion group, the volume of cerebral infarction in the ischemic preconditioning group was reduced, and the difference was statistically significant (P0.05).3.HE staining showed that the nerve cells in the sham operation group were not injured; and the ischemia reperfusion group was in the ischemic reperfusion group. Compared with the ischemic preconditioning group, the damage of the hippocampal neurons in the ischemic preconditioning group was significantly enhanced, compared with the sham group. The expression of Beclin-1 in the ischemic preconditioning group was significantly enhanced, and the difference was statistically significant (P0.05). Compared with the ischemic reperfusion group, the number of Beclin-1 positive cells decreased and the difference was statistically significant compared with the ischemic reperfusion group. The difference was statistically significant (P0.05). The expression of a very small number of LC3-II positive cells was found in the sham operation group of P0.05.5., while a large number of LC3-II positive cells were found in the ischemic reperfusion group and the ischemic preconditioning group. The difference was statistically significant (P0.05). The expression of LC3-II positive cells in each subgroup of ischemic preconditioning was significantly lower than that of the ischemia reperfusion group, and the difference was statistically significant (P0.05).6. transmission electricity The number of autophagosomes and autophagic lysosomes in the ischemic preconditioning group and ischemia reperfusion group were in different periods. The number of the ischemic preconditioning group was significantly reduced by the.7. ischemic preconditioning 72h subgroup, compared with the 24h subgroup and the 5D subgroup, the cerebral infarction volume decreased (P0.05), the hippocampal neuronal damage was reduced, Beclin-1, LC3-II positive were fine. The number of cells decreased (P0.05), autophagosome formation and lysosome activation decreased.
Conclusion 1. the model of ischemic preconditioning and ischemia-reperfusion model of middle cerebral artery made by modified Zea-Longa thread embolus method is simple and safe. The data reliable.2. ischemic preconditioning can reduce the neurological dysfunction score of rats after 24 hours of ischemia reperfusion, the volume of cerebral infarction, the damage of the hippocampal nerve cells, and the fine nerve after ischemia-reperfusion. .3. ischemic preconditioning may reduce the autophagic related protein Beclin-1, LC3-II expression after 24 hours of ischemia-reperfusion, reduce the autophagic death of ischemic reperfusion injury, reduce the symptoms of nerve function defect, reduce the volume of cerebral infarction, and thus play its protective role of.4. ischemic preconditioning 72h The volume of cerebral infarction in subgroup was lower than that of ischemic preconditioning (24h), 5D subgroup, Beclin-1, LC3-II positive cells were lower than that of ischemic preconditioning 24h, 5D subgroup and autophagosome, and the number of lysosomes at all levels was relatively small, suggesting that the best preconditioning time interval was 72h..

【學位授予單位】：河北聯(lián)合大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R743.3

【參考文獻】

相關期刊論文 前10條

1 乜全民;張世明;;大鼠局灶性腦缺血再灌注損傷后自噬的初步研究[J];中國臨床神經科學;2010年01期

2 吳淑燕;李瓊;儲元元;李Z腦

本文編號：1814821