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

【摘要】：目的研究自噬相關(guān)蛋白Beclin-1、LC3-II在缺血預(yù)處理后不同時(shí)間間隔大鼠腦缺血再灌注海馬中的表達(dá)，探討自噬在腦缺血預(yù)處理神經(jīng)保護(hù)作用的可能機(jī)制。方法1.健康雄性SD大鼠50只隨機(jī)分為3組：假手術(shù)組、缺血再灌注組、缺血預(yù)處理組，缺血預(yù)處理組再按不同缺血預(yù)處理間隔時(shí)間分為24h、72h、5d3個(gè)亞組。2.參照Zea-Longa線栓法制作大鼠大腦中動(dòng)脈缺血預(yù)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(xué)染色、觀察腦梗死體積、海馬神經(jīng)細(xì)胞病理變化、海馬神經(jīng)細(xì)胞自噬相關(guān)蛋白Beclin-1及LC3-II表達(dá)的情況。4.透射電鏡觀察神經(jīng)細(xì)胞中自噬超微結(jié)構(gòu)的變化。5.比較各組神經(jīng)功能缺失、腦梗死體積、海馬神經(jīng)細(xì)胞病理變化、海馬神經(jīng)細(xì)胞自噬相關(guān)蛋白Beclin-1及LC3-II表達(dá)、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(jì)學(xué)軟件進(jìn)行數(shù)據(jù)分析，所得數(shù)據(jù)均以x±s表示，P0.05認(rèn)為差異有統(tǒng)計(jì)學(xué)意義。采用Microsoft Excel軟件作圖。 結(jié)果1.缺血預(yù)處理組神經(jīng)功能缺失癥狀較缺血再灌注組有所改善，差異具有統(tǒng)計(jì)學(xué)意義（P0.05）。2.與缺血再灌注組相比，缺血預(yù)處理組腦梗死體積縮小，差異具有統(tǒng)計(jì)學(xué)意義（P0.05）。3.HE染色可見假手術(shù)組神經(jīng)細(xì)胞無損傷；與缺血再灌注組相比，缺血預(yù)處理各亞組海馬神經(jīng)細(xì)胞損傷減輕。4.與假手術(shù)組相比，缺血再灌注組、缺血預(yù)處理組Beclin-1的表達(dá)均明顯增強(qiáng)，差異具有統(tǒng)計(jì)學(xué)意義（P0.05）；缺血預(yù)處理組與缺血再灌注組比較，可見Beclin-1陽性細(xì)胞數(shù)減少，差異具有統(tǒng)計(jì)學(xué)意義（P0.05）。5.假手術(shù)組只見極少量LC3-II陽性細(xì)胞表達(dá)，而在缺血再灌注組及缺血預(yù)處理組均可見大量LC3-II陽性細(xì)胞，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。缺血預(yù)處理各亞組LC3-II陽性細(xì)胞表達(dá)顯著低于缺血再灌注組，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。6.透射電鏡下缺血預(yù)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(yù)處理組明顯減少。7.缺血預(yù)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(jīng)細(xì)胞損傷減輕，Beclin-1、LC3-II陽性細(xì)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(jié)論1.采用改良后Zea-Longa線栓法制作的大鼠大腦中動(dòng)脈缺血預(yù)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(yù)處理可減低缺血再灌注24小時(shí)后大鼠神經(jīng)功能障礙評分、腦梗死體積、海馬神經(jīng)細(xì)胞損傷，對缺血再灌注后神經(jīng)細(xì)胞損傷具有保護(hù)作用。3.缺血預(yù)處理可能通過下調(diào)缺血再灌注24小時(shí)后自噬相關(guān)蛋白Beclin-1、LC3-II的表達(dá)，以減少缺血再灌注損傷神經(jīng)細(xì)胞的自噬性死亡，減輕神經(jīng)功能缺損癥狀，縮小腦梗死體積，從而發(fā)揮其腦保護(hù)作用。4.缺血預(yù)處理72h亞組腦梗死體積低于缺血預(yù)處理24h、5d亞組，Beclin-1、LC3-II陽性細(xì)胞數(shù)低于缺血預(yù)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(yù)處理誘導(dǎo)時(shí)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(xué)位授予單位】：河北聯(lián)合大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R743.3

【參考文獻(xiàn)】

相關(guān)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(jīng)科學(xué);2010年01期

2 吳淑燕;李瓊;儲元元;李Z腦

本文編號：1814821