本文選題：自噬　切入點(diǎn)：腦創(chuàng)傷　出處：《華北理工大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：目的探索腦創(chuàng)傷后的自噬水平神經(jīng)突觸再生之間的關(guān)系。從本實(shí)驗(yàn)通過應(yīng)用自噬激動(dòng)劑辛伐他汀和自噬抑制劑氯喹對腦創(chuàng)傷的大鼠進(jìn)行干預(yù),檢查藥物對大鼠海馬區(qū)的自噬相關(guān)蛋白LC3和Beclin-1的表達(dá)水平的影響,以PSD-95和synaptophysin指示突觸密度的高低。研究腦創(chuàng)傷后神經(jīng)功能回復(fù)的機(jī)制,為腦創(chuàng)傷后臨床治療提供新的治療靶點(diǎn)。方法按隨機(jī)原則將200只Sprague Dawley雄性大鼠分為4組:1 Sham組(n=50)2TBI組(n=50)3辛伐他汀組(n=50)4氯喹組(n=50)。每組按手術(shù)后時(shí)間1d,3d,5d及7d分成四個(gè)亞組。使用Marmarou法,使用高空墜物打擊的方法,建立大鼠彌漫性腦損傷模型。并進(jìn)行以下各項(xiàng)檢查:1使用HE染色法顯示海馬區(qū)神經(jīng)元的形態(tài);2免疫熒光檢測LC3與Neu N的表達(dá)定位情況;3蛋白質(zhì)印記法檢測自噬相關(guān)蛋白LC3,Beclin-1和突觸蛋白PSD-95,synaptophysin的表達(dá)水平。4 50只SD雄性大鼠在創(chuàng)傷后3-9d行Morris水迷宮測試。用q檢驗(yàn)比較兩組間的數(shù)據(jù),組間使用單因素方差分析,P0.05作為差異具有統(tǒng)計(jì)學(xué)意義的界限。結(jié)果1 HE染色結(jié)果。sham組:海馬神經(jīng)元無明顯的病理學(xué)改變,神經(jīng)元清晰,整齊。TBI組:神經(jīng)元數(shù)目減少,排列稀疏,水腫,細(xì)胞周圍間隙和血管周圍間隙較sham組變寬。辛伐他汀組:神經(jīng)元數(shù)量減少較TBI組輕,輕微水腫,細(xì)胞周圍間隙和血管周圍間隙變寬較TBI組輕。氯喹組:與TBI組相比沒有明顯區(qū)別。2大鼠的空間學(xué)習(xí)記憶功能檢測。TBI組在傷后的各組行為學(xué)參數(shù),逃避潛伏期變長,穿越平臺次數(shù),平臺所在象限路程比及時(shí)間比均顯著減少。辛伐他汀組逃避潛伏期較TBI組變短,穿越平臺次數(shù),平臺所在象限路程比及時(shí)間比均顯著增加。氯喹組逃避潛伏期長于TBI組,穿越平臺次數(shù),平臺所在象限路程比及時(shí)間比均減少。3 LC3 II的Western blot檢測結(jié)果。Sham組:只可以檢測到輕微量的蛋白。TBI組:LC3 II的蛋白表達(dá)量與Sham組比較,傷后1d出現(xiàn)明顯升高(P0.05),3d達(dá)到表達(dá)最高點(diǎn),創(chuàng)傷后7d的表達(dá)依然高于Sham組(P0.05)。辛伐他汀組:LC3 II表達(dá)創(chuàng)傷后1d和3d的表達(dá)量明顯高于TBI組,有統(tǒng)計(jì)學(xué)意義(P0.05)。氯喹組:LC3 II蛋白表達(dá)TBI組相比較,在傷后3d和5d表達(dá)較低,具有統(tǒng)計(jì)學(xué)意義(P0.05)。4 Beclin-1的Western blot檢測結(jié)果。Sham組:只可以檢測到輕微量的蛋白,在各時(shí)像點(diǎn)無差異。TBI組:Beclin-1的蛋白表達(dá)量與Sham組比較,傷后1d出現(xiàn)明顯升高(P0.05),3d達(dá)到表達(dá)最高點(diǎn),創(chuàng)傷后7d的表達(dá)依然高于Sham組(P0.05)。辛伐他汀組:Beclin-1表達(dá)隨時(shí)間變化的趨勢和TBI組類似,在創(chuàng)傷后1d,3d和5d的表達(dá)量明顯高于TBI組,有統(tǒng)計(jì)學(xué)意義(P0.05)。氯喹組:Beclin-1蛋白表達(dá)與時(shí)間的對應(yīng)關(guān)系和TBI組類似,與TBI組相比較,在傷后3d表達(dá)較低,具有統(tǒng)計(jì)學(xué)意義(P0.05)。5 PSD-95的Western blot檢測結(jié)果。Sham組:PSD-95蛋白表十分達(dá)明顯,在各時(shí)像點(diǎn)無差異。TBI組:PSD-95的蛋白表達(dá)量與Sham組比較明顯降低,之后幾個(gè)時(shí)間點(diǎn),PSD-95的表達(dá)會(huì)有緩慢的上升,但在7d依然明顯低于sham組(P0.05)。辛伐他汀組:PSD-95表達(dá)隨時(shí)間變化的趨勢和TBI組類似,與TBI組相比,5d和7d的差別有統(tǒng)計(jì)學(xué)意義(P0.05)。氯喹組:PSD-95蛋白表達(dá)與TBI組類似,更為緩慢,在傷后7d表達(dá)較低,具有統(tǒng)計(jì)學(xué)意義(P0.05)。6 synaptophysin的Western blot檢測結(jié)果。Sham組:synaptophysin蛋白表達(dá)十分明顯,各時(shí)像點(diǎn)無差異。TBI組:synaptophysin的蛋白表達(dá)量與Sham組比較明顯降低,之后幾個(gè)時(shí)間點(diǎn),synaptophysin的表達(dá)會(huì)有緩慢的上升,但在7d依然明顯低于sham組(P0.05)。辛伐他汀組:synaptophysin表達(dá)隨時(shí)間變化的趨勢和TBI組類似,與TBI組相比,5d和7d的差別有統(tǒng)計(jì)學(xué)意義(P0.05)。氯喹組:synaptophysin蛋白表達(dá)與時(shí)間的對應(yīng)關(guān)系和TBI組類似,與TBI組相比較,在傷后5d和7d較低,具有統(tǒng)計(jì)學(xué)意義(P0.05)。7 LC3與Neu N的免疫熒光檢測結(jié)果。紅色熒光顯示自噬標(biāo)記蛋白LC3的表達(dá),綠色熒光顯示神經(jīng)元標(biāo)記物Neu N的表達(dá)。結(jié)論辛伐他汀對大鼠的彌漫性腦創(chuàng)傷有保護(hù)作用,可以顯著地提高神經(jīng)功能的回復(fù),海馬區(qū)的突觸相關(guān)蛋白表達(dá)水平提高。其可能的機(jī)制是通過加強(qiáng)自噬的水平,上調(diào)了神經(jīng)網(wǎng)絡(luò)的可塑性,使更多的突觸得以重建,神經(jīng)網(wǎng)絡(luò)的完整性得到了更好的重建,而大鼠的學(xué)習(xí)記憶功能也得到了改善。
[Abstract]:Objective to explore the relationship between the level of autophagy in synaptic regeneration after traumatic brain injury. The intervention from the experiment on traumatic brain injury by using autophagy agonist simvastatin and chloroquine in rats, to examine the effects of drugs on rat hippocampus autophagy related protein LC3 and Beclin-1 expression, with PSD-95 and synaptophysin indicating synapse density of the mechanism. Reply to neural function after traumatic brain injury, for the treatment of brain trauma provide new therapeutic targets. Methods according to the principle of random 200 Sprague male Dawley rats were divided into 4 groups: 1 in group Sham (n=50) 2TBI group (n=50) 3 (n=50 4) simvastatin group chloroquine group (n=50). Each group according to the postoperative time of 1D, 3D, 5D and 7d are divided into four sub groups. Using the Marmarou method, using the method of falling down, establish the model of diffuse brain injury in rats. And the following tests: 1 using HE staining method According to the morphology of hippocampal neurons; localization of immunofluorescence detection of LC3 and Neu 2 N; 3 protein blot analysis of autophagy related protein LC3, Beclin-1 and synaptic protein PSD-95, synaptophysin expression level of.4 50 SD male rats at 3-9d after trauma underwent Morris water maze test. Using Q test to compare between the two groups data analysis using single factor variance between groups, as the limits of P0.05. The difference was statistically significant. The 1 group.Sham: the results of HE staining in hippocampal neurons without obvious pathological changes of neurons in.TBI group: clear, neat and reduce the number of neurons, sparse, edema, and blood vessel cells around the gap gap compared with the sham group variable wide. Simvastatin group: reduce the number of neurons was less than that in TBI group, mild edema, clearance and vascular cells around the gap becomes wider than the TBI group of light. There is no obvious difference between chloroquine group:.2 rats compared with TBI group The spatial learning and memory function of each behavior detection of group.TBI after injury the parameters, the escape latency became longer, the number of crossing platform, platform quadrant distance and time were reduced significantly. In simvastatin group compared with TBI group, the escape latency shortened, the times of crossing the platform, the platform quadrant time and distance were significantly increased than chloroquine group escape. The incubation period is longer than that of group TBI, the number of crossing platform, platform quadrant distance and time ratio were reduced by Western blot.3 LC3 II.Sham test results: group.TBI protein can be detected only a trace of light: the expression of LC3 II protein compared with Sham group, 1D increased significantly after injury (P0.05), 3D expression the highest point, the expression of 7D after trauma is still higher than that of Sham group (P0.05). Simvastatin group: LC3 II expression after trauma and expression of 1D 3D was significantly higher than that in TBI group, there was statistical significance (P0.05). Chloroquine group: LC3 II protein The expression of TBI compared to the group, at 3D after injury and the expression of 5D was lower, with statistical significance (P0.05) Western blot.4 Beclin-1.Sham group test results: only can detect light trace protein at different time points as there was no difference between group.TBI: the expression quantity of Beclin-1 compared with Sham group, 1D after injury there were significantly increased (P0.05), 3D expression reached the highest point, the expression of 7D after trauma is still higher than that of Sham group (P0.05). Simvastatin group: Beclin-1 expression trends over time and is similar to that of group TBI at 1D after trauma, the expression of 3D and 5D was significantly higher than that in group TBI, with statistical significance (P0.05). Chloroquine group: time and corresponding relation and similar to the TBI group of Beclin-1 protein expression, compared with the TBI group, the expression of 3D after injury was lower, with statistical significance (P0.05) Western blot.5 PSD-95.Sham group test results: PSD-95 protein was significantly different in the table is like a point, there was no difference between group.TBI: PSD-95 The amount of Sham group decreased significantly compared with the expressed protein, after some time point, the expression of PSD-95 is a slow rise in 7d, but still significantly lower than sham group (P0.05). Simvastatin group: PSD-95 expression trends over time and similar to the TBI group, compared with the TBI group, there was statistical significance in 5D and 7d the difference (P0.05). Chloroquine group: the expression of PSD-95 protein is similar with the TBI group, more slowly, after injury 7d expression was lower, with statistical significance (P0.05) Western blot.6 synaptophysin group.Sham results: the expression of synaptophysin is very obvious, like the point of no difference in.TBI group decreased significantly compared with the amount of the expression of synaptophysin protein in Sham group, after some time point, the expression of synaptophysin is a slow rise in 7d, but still significantly lower than sham group (P0.05). Simvastatin group: synaptophysin expression trends over time and is similar to that of group TBI and TBI Compared with statistical significance of 5D and 7d (P0.05). The difference between chloroquine group: time and corresponding relationship between TBI group and similar synaptophysin protein expression, compared with TBI, 5D and 7d after injury was lower, with statistical significance (P0.05) immunofluorescence detection results of.7 LC3 and Neu N. The red fluorescence showed that the expression of autophagy marker protein LC3, green fluorescence showed that the expression of neuronal markers Neu N. Conclusion simvastatin in rats with diffuse brain injury has a protective effect, can significantly improve the neurological function recovery, the expression level of synapse related protein in hippocampus increased. The possible mechanism is through enhancing the level of autophagy that raised the plasticity of neural network, the more able to reconstruct the synaptic integrity, the neural network has better reconstruction, and learning and memory function of rats was improved.

【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R651.15

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 云中客;;對腦創(chuàng)傷的物理探討[J];物理;2009年11期

2 劉興波,衣服新,羅俊生,王秋華;二次腦創(chuàng)傷臨床診斷指標(biāo)的初步探討[J];錦州醫(yī)學(xué)院學(xué)報(bào);2002年03期

3 趙景霞,劉清軍,崔建忠,洪軍,宋朝彥;大鼠腦創(chuàng)傷后運(yùn)動(dòng)功能障礙及美洛寧對其作用機(jī)理的研究[J];四川大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2003年03期

4 雪亮,楊樹源;人腦創(chuàng)傷后神經(jīng)元凋亡及調(diào)節(jié)機(jī)制的觀察[J];中華創(chuàng)傷雜志;2003年03期

5 賈叢林;腦源性神經(jīng)營養(yǎng)因子在腦創(chuàng)傷中表達(dá)的動(dòng)物實(shí)驗(yàn)研究[J];河南實(shí)用神經(jīng)疾病雜志;2004年02期

6 楊樹源,楊新宇;急性腦創(chuàng)傷后繼發(fā)性神經(jīng)細(xì)胞損傷的研究進(jìn)展[J];中華神經(jīng)外科雜志;2004年02期

7 蔣平,呂軍,焦炳華;基因芯片技術(shù)在腦創(chuàng)傷研究中的應(yīng)用[J];醫(yī)學(xué)綜述;2004年12期

8 黃巨恩,楊志平,陳紆,曾慶堂;堿性成纖維細(xì)胞生長因子對腦創(chuàng)傷后機(jī)體免疫功能的影響[J];廣西醫(yī)科大學(xué)學(xué)報(bào);2004年06期

9 陳瑋,李堯;充足的碳水化合物對腦創(chuàng)傷患者的意義[J];職業(yè)與健康;2005年08期

10 趙紅崗;李東飛;李東亮;田梅;張耀東;李慶崗;李明陽;;黃體酮對大鼠腦創(chuàng)傷后神經(jīng)干細(xì)胞增殖的影響(英文)[J];中國臨床康復(fù);2006年33期

相關(guān)會(huì)議論文 前10條

1 楊樹源;楊新宇;;急性腦創(chuàng)傷后繼發(fā)性神經(jīng)細(xì)胞損傷的研究進(jìn)展[A];中國醫(yī)師協(xié)會(huì)神經(jīng)外科醫(yī)師分會(huì)成立大會(huì)資料匯編[C];2004年

2 邱仲慶;;腦低溫術(shù)對腦創(chuàng)傷時(shí)腦缺血及損傷之療效[A];第六屆海峽兩岸心血管科學(xué)研討會(huì)論文摘要集[C];2007年

3 姚世斌;李安民;梁樹立;趙明;張宏達(dá);;腦創(chuàng)傷致遲發(fā)性腦梗塞臨床及相關(guān)因素分析[A];中國醫(yī)師協(xié)會(huì)神經(jīng)外科醫(yī)師分會(huì)第二屆全國代表大會(huì)論文匯編[C];2007年

4 江榮才;栗站營;王彬;陳潔麗;張建寧;;孕酮增強(qiáng)了老年腦創(chuàng)傷大鼠循環(huán)血內(nèi)皮祖細(xì)胞動(dòng)員、腦血管再生和神經(jīng)功能修復(fù)[A];2011中華醫(yī)學(xué)會(huì)神經(jīng)外科學(xué)學(xué)術(shù)會(huì)議論文匯編[C];2011年

5 陳勝咸;林茂村;張峰銘;;雌激素治療實(shí)驗(yàn)性腦創(chuàng)傷:血管內(nèi)皮細(xì)胞生長因子參與(英文)[A];第七屆海峽兩岸心血管科學(xué)研討會(huì)論文集[C];2009年

6 張賽;;基質(zhì)衍生因子-1與干細(xì)胞的腦創(chuàng)傷趨向性機(jī)制分析[A];中國醫(yī)師協(xié)會(huì)神經(jīng)外科醫(yī)師分會(huì)第四屆全國代表大會(huì)論文匯編[C];2009年

7 常小麗;黃慧玲;莫麗冬;范維嘉;王辰;;�；撬釋δX創(chuàng)傷大鼠腦生化代謝指標(biāo)的影響[A];第七屆全國創(chuàng)傷學(xué)術(shù)會(huì)議暨2009海峽兩岸創(chuàng)傷醫(yī)學(xué)論壇論文匯編[C];2009年

8 王彬;孫林;栗戰(zhàn)營;張建寧;江榮才;;阿托伐他汀對腦創(chuàng)傷大鼠內(nèi)皮祖細(xì)胞和血管新生的實(shí)驗(yàn)研究[A];2011中華醫(yī)學(xué)會(huì)神經(jīng)外科學(xué)學(xué)術(shù)會(huì)議論文匯編[C];2011年

9 鄧杰;王明明;方國安;金秀國;;大黃對腦創(chuàng)傷患者血清細(xì)胞因子的早期影響[A];2004年浙江省危重病學(xué)學(xué)術(shù)年會(huì)論文匯編[C];2004年

10 陳勝咸;;雌激素可抑制腦創(chuàng)傷引致的腦血管功能異常[A];第六屆海峽兩岸心血管科學(xué)研討會(huì)論文摘要集[C];2007年

相關(guān)重要報(bào)紙文章 前2條

1 莫秋;提高重型腦創(chuàng)傷療效[N];科技日報(bào);2005年

2 劉莉莉;“沖擊波”戰(zhàn)爭：可能讓兩萬英軍受創(chuàng)傷[N];新華每日電訊;2007年

相關(guān)博士學(xué)位論文 前10條

1 王晨;急性腦創(chuàng)傷后人腦皮層蛋白質(zhì)組表達(dá)變化的研究[D];天津醫(yī)科大學(xué);2005年

2 羅成義;腦創(chuàng)傷后絲裂原活化蛋白激酶信號轉(zhuǎn)導(dǎo)的作用及調(diào)控機(jī)理[D];第一軍醫(yī)大學(xué);2001年

3 雷平;基因芯片篩選人腦創(chuàng)傷皮層差異表達(dá)基因的實(shí)驗(yàn)研究[D];天津醫(yī)科大學(xué);2006年

4 郭新賓;內(nèi)皮祖細(xì)胞和大鼠腦創(chuàng)傷后損傷組織再生修復(fù)關(guān)系的實(shí)驗(yàn)研究[D];天津醫(yī)科大學(xué);2010年

5 蔣平;應(yīng)用組織芯片對腦創(chuàng)傷下小膠質(zhì)細(xì)胞凋亡的研究及小膠質(zhì)細(xì)胞凋亡相關(guān)蛋白的蛋白質(zhì)組學(xué)研究[D];第二軍醫(yī)大學(xué);2005年

6 梁海乾;骨橋蛋白在大鼠腦創(chuàng)傷中的作用研究[D];天津醫(yī)科大學(xué);2008年

7 唐兆華;C-EPO對創(chuàng)傷性腦水腫的保護(hù)作用及機(jī)制探討[D];重慶醫(yī)科大學(xué);2013年

8 瞿文軍;腦創(chuàng)傷后兒茶酚胺遞質(zhì)對神經(jīng)細(xì)胞損害的線粒體調(diào)控機(jī)制的實(shí)驗(yàn)研究[D];第一軍醫(yī)大學(xué);2004年

9 劉慶國;辛伐他汀對大鼠腦創(chuàng)傷后內(nèi)皮祖細(xì)胞動(dòng)員和損傷腦組織血管再生影響的實(shí)驗(yàn)研究[D];天津醫(yī)科大學(xué);2009年

10 李愛林;重型腦創(chuàng)傷后細(xì)胞膜損傷及亞低溫對其影響的研究[D];天津醫(yī)科大學(xué);2006年

相關(guān)碩士學(xué)位論文 前10條

1 王勇朝;BBG對大鼠腦創(chuàng)傷后炎性反應(yīng)的抑制作用及機(jī)制研究[D];河北聯(lián)合大學(xué);2014年

2 張弘翱;辛伐他汀及氯喹調(diào)節(jié)大鼠腦創(chuàng)傷后突觸重建的研究[D];華北理工大學(xué);2015年

3 常小麗;�；撬釋χ匦湍X創(chuàng)傷大鼠腦代謝及線粒體作用研究[D];天津醫(yī)科大學(xué);2010年

4 王曉楠;重組人粒細(xì)胞集落刺激因子動(dòng)員內(nèi)皮祖細(xì)胞對腦創(chuàng)傷大鼠功能恢復(fù)的影響[D];天津醫(yī)科大學(xué);2010年

5 霍永強(qiáng);廓清療法治療實(shí)驗(yàn)性腦創(chuàng)傷[D];廣西醫(yī)科大學(xué);2007年

6 張雷;腦創(chuàng)傷兒童顱內(nèi)壓及腦灌注壓的臨床特點(diǎn)分析[D];重慶醫(yī)科大學(xué);2014年

7 陳興河;腦創(chuàng)傷后能量代謝的變化及尼莫地平對其影響的實(shí)驗(yàn)研究[D];天津醫(yī)科大學(xué);2003年

8 栗戰(zhàn)營;黃體酮促進(jìn)老年腦創(chuàng)傷大鼠血管再生和神經(jīng)再生[D];河北聯(lián)合大學(xué);2011年

9 梁平;鹽酸納洛酮對大鼠腦創(chuàng)傷后細(xì)胞免疫功能影響的實(shí)驗(yàn)研究[D];重慶醫(yī)科大學(xué);2006年

10 張鵬;人臍血間充質(zhì)干細(xì)胞移植治療腦創(chuàng)傷大鼠的實(shí)驗(yàn)研究[D];鄭州大學(xué);2005年

，

本文編號：1631994