本文選題：缺血后處理　切入點(diǎn)：血腦屏障　出處：《吉林大學(xué)》2015年博士論文　論文類型：學(xué)位論文

【摘要】：腦血管病是一種嚴(yán)重危害人類健康的疾病，，已成為全球人類死亡的三大原因之一。其中因頸動(dòng)脈狹窄性疾病引發(fā)的卒中占所有卒中的1/4~1/2。在美國(guó)幾乎1/3的卒中來源于頸動(dòng)脈狹窄，而在我國(guó)來源于顱外頸動(dòng)脈狹窄的缺血性腦卒中的年發(fā)病率為120~180/10萬，年病死率更是可達(dá)80~120/10萬。目前頸動(dòng)脈狹窄的治療方法有藥物治療、手術(shù)治療等。但就現(xiàn)在來看，重度狹窄頸動(dòng)脈早期的血管重建能夠有效的減少卒中的復(fù)發(fā)率，降低死亡率。雖然血管重建能解決以上問題，但血管重建之后的并發(fā)癥例如神經(jīng)認(rèn)知功能障礙、嚴(yán)重的腦水腫、持續(xù)頭痛、顱內(nèi)出血，甚至死亡嚴(yán)重威脅患者生存狀態(tài)。因此尋找有效的方法解決血管重建之后的并發(fā)癥已成為一個(gè)亟待解決的問題。 血腦屏障由膠質(zhì)細(xì)胞、周細(xì)胞、血管內(nèi)皮細(xì)胞和細(xì)胞外間隙組成，是一個(gè)介于血液和腦組織之間的復(fù)雜的系統(tǒng)，也是神經(jīng)系統(tǒng)特有的結(jié)構(gòu)。在血腦屏障結(jié)構(gòu)和功能完整的情況下，血腦屏障可阻擋某些大分子物質(zhì)通過并對(duì)血液和腦組織之間的物質(zhì)交換進(jìn)行嚴(yán)格的篩選。而在血腦屏障功能和結(jié)構(gòu)遭到破壞時(shí)，血腦屏障通透性增高，腦水腫形成。血腦屏障正常通透性主要靠血管內(nèi)皮細(xì)胞之間的緊密連接（Tight junction,TJ）來維持。而緊密連接主要有緊密連接蛋白ZO，occludin，跨膜蛋白，claudins和連接黏附分子（Juntional adhesion molecule，JAM）構(gòu)成。其中occludin和claudin-5是形成緊密連接的關(guān)鍵蛋白。它們的表達(dá)和分布變化可導(dǎo)致血腦屏障的完整性發(fā)生變化。除此之外與血腦屏障的完整性密切相關(guān)的還有基質(zhì)金屬蛋白酶家族。其中MMP-2、MMP-9在基質(zhì)金屬蛋白酶家族中占有重要地位。它們主要作用是水解細(xì)胞基質(zhì)和基膜的主要成分Ⅳ型膠原。已有研究證明MMP-2、MMP-9與血腦屏障結(jié)構(gòu)和功能的變化有密切的關(guān)系，尤其MMP-9能夠通過降解claudin-5和occludin等來調(diào)節(jié)血腦屏障的通透性。因此我們重點(diǎn)研究MMP-9、claudin-5和occludin與血腦屏障功能、腦水腫之間的關(guān)系，并進(jìn)一步明確蛋白之間的相互作用。 目前，在多種器官中，缺血后處理能夠減輕缺血/再灌注帶來的損傷。其方法是再灌注的早期快速的、間歇性的阻斷血流進(jìn)而改變血流的動(dòng)力學(xué)水平。雖然以往的實(shí)驗(yàn)中我們已經(jīng)證明缺血后處理能夠阻止早期解除頸動(dòng)脈狹窄所導(dǎo)致的神經(jīng)元延遲性死亡，但缺血后處理對(duì)頸動(dòng)脈狹窄解除后的大腦水腫和血腦屏障破壞的影響及其機(jī)制尚不清楚。 實(shí)驗(yàn)?zāi)康模?在大鼠重度頸動(dòng)脈狹窄模型中探討缺血后處理對(duì)頸動(dòng)脈狹窄解除引起的腦水腫和血腦屏障破壞的影響及其作用機(jī)制。 實(shí)驗(yàn)方法: 通過兩側(cè)頸動(dòng)脈與不銹鋼微管綁定建立Wistar大鼠低灌注模型。缺血后處理步驟包括30秒缺血/30秒再灌注，循環(huán)三次。通過測(cè)量腦組織含水量對(duì)腦水腫的程度進(jìn)行評(píng)估，并且通過檢測(cè)伊文思藍(lán)(EB)和熒光素鈉（NaF）的在腦組織的含量來評(píng)估血腦屏障通透性的變化。分別用明膠酶譜分析和原位酶譜分析檢測(cè)MMP-9的活性和定位。用免疫組織化學(xué)觀察緊密連接蛋白claudin-5和occludin的分布。 實(shí)驗(yàn)結(jié)果： （1）血腦屏障通透性研究顯示頸動(dòng)脈狹窄解除組與假手術(shù)組和狹窄組相比，腦組織含水量增加，EB、NaF在大腦中的濃度顯著提高，但這種變化被缺血后處理減弱。 （2）明膠酶譜和熒光原位明膠酶譜研究顯示MMP-9主要分布在皮層并且頸動(dòng)脈狹窄的解除能導(dǎo)致MMP-9活性的升高，但這種活性的升高能被缺血后處理顯著抑制。 （3）免疫組織化學(xué)揭示了缺血后處理提高了claudin-5和occlaudin的連續(xù)分布。 （4）ELISA檢測(cè)顯示由于頸動(dòng)脈狹窄解除導(dǎo)致MMP-9的表達(dá)上調(diào)，進(jìn)行缺血后處理后以上變化減弱，但是頸動(dòng)脈狹窄解除后claudin-5和occlaudin的表達(dá)下調(diào)，進(jìn)行缺血后處理后這兩種蛋白的表達(dá)上調(diào)。 實(shí)驗(yàn)結(jié)論： 缺血后處理是一種有效防止腦水腫和改善血腦屏障通透性的方法，并且能夠在重度頸動(dòng)脈狹窄的解除過程中應(yīng)用。
[Abstract]:Cerebrovascular disease is a serious hazard to human health disease, has become one of the three major causes of global human death. The stroke caused by carotid artery stenosis disease stroke stroke accounted for all sources of 1/4~1/2. in the United States almost 1/3 in carotid artery stenosis and ischemic stroke in our country from the extracranial carotid artery narrow the annual incidence rate of 120~180/10 million, the annual mortality rate is up to 80~120/10 million. The treatment of carotid artery stenosis with drug therapy, surgical treatment and so on. But now, severe stenosis of carotid artery early revascularization can reduce the effective stroke recurrence rate, reduce mortality. Although vascular reconstruction to solve the above problems, but after the reconstruction of vascular complications such as neurocognitive dysfunction, severe brain edema, persistent headache, intracranial hemorrhage, and even death serious threat to patient survival. Because of this search Finding effective methods to solve the complications after vascular reconstruction has become an urgent problem.
The blood brain barrier by glial cells, pericytes, endothelial cells and extracellular space, is a complex system between blood and brain tissue, is unique to the nervous system structure. In the structure and function of blood-brain barrier integrity under the condition of the blood brain barrier can block some macromolecules through and on between blood and brain tissue material exchange strict screening. The blood brain barrier and the destruction of the structure, increase the permeability of blood brain barrier and brain edema formation. The normal blood brain barrier permeability mainly depends on the vascular endothelial cell tight junctions (Tight junction, TJ) to maintain and are closely connected closely. Connected proteins ZO, occludin, a transmembrane protein, claudins and junctional adhesion molecules (Juntional adhesion, molecule, JAM). Occludin and claudin-5 is closely connected with the expression of key proteins. The distribution and changes to the integrity of the blood-brain barrier changes. And matrix metalloproteinase family integrity besides and blood brain barrier are closely related. The MMP-2, MMP-9 plays an important role in the matrix metalloproteinase family. Their main function is the main component of type IV collagen hydrolysis of cell matrix and basement membrane. The research proved that MMP-2, there is a close relationship between MMP-9 and changes of blood brain barrier structure and function, especially MMP-9 through degradation of claudin-5 and occludin to regulate the permeability of the blood-brain barrier. So we focus on the study of MMP-9, claudin-5 and occludin and blood brain barrier function, the relationship between brain edema, and further clarify the interaction between proteins role.
At present, in a variety of organs, postprocessing can reduce the damage of ischemia and ischemia / reperfusion brings. The method is fast in the early period of reperfusion, intermittent occlusion of blood flow and change the blood flow dynamics. Although previous experiments we have shown that ischemic postprocessing can prevent early termination of carotid artery stenosis caused by delayed neuron death, but ischemic postprocessing effects on carotid artery stenosis after relieving brain edema and blood-brain barrier damage and its mechanism is not clear.
Objective:
The effects and mechanisms of ischemic postconditioning on cerebral edema and blood-brain barrier damage induced by carotid artery stenosis were studied in a rat model of severe carotid stenosis.
Experimental methods:
Through both sides of the carotid artery and the stainless steel tube binding model of Wistar rat ischemia reperfusion model. Low postprocessing steps include 30 seconds /30 seconds after ischemia reperfusion, three cycles. By measuring the brain water content on the degree of cerebral edema were evaluated, and detected by Evans blue (EB) and fluorescein sodium (NaF) in the brain the content of organization to assess the changes of blood-brain barrier permeability respectively. The spectrum analysis of the activity and location of detection of MMP-9 by gelatin zymography and in situ enzyme distribution. By immunohistochemical observation of tight junction protein claudin-5 and occludin.
Experimental results:
(1) blood brain barrier permeability study showed that compared with sham operation group and stenosis group, the water content of brain tissue increased, and the concentration of EB and NaF increased significantly in the carotid artery stenosis relieving group, but this change was weakened by ischemic postconditioning.
(2) gelatin zymography and fluorescence in situ gelatinase analysis showed that MMP-9 was mainly distributed in the cortex, and the release of carotid stenosis could lead to the increase of MMP-9 activity. However, the increase of this activity could be significantly inhibited by ischemic postconditioning.
(3) immuno histochemistry revealed that post ischemic treatment enhanced the continuous distribution of claudin-5 and occlaudin.
(4) ELISA detection showed that the expression of MMP-9 was upregulated due to the release of carotid artery stenosis, and the above changes decreased after ischemic postconditioning. However, the expression of claudin-5 and occlaudin was down regulated after carotid artery stenosis, and the expression of these two proteins increased after ischemia.
Experimental conclusions:
Ischemic postconditioning is an effective method to prevent brain edema and improve the permeability of the blood brain barrier, and can be used in the process of severe carotid stenosis.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R743.31

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 洪波,劉建民,許奕,趙文元,黃清海,辛濤,張瓏,張?chǎng)?血栓保護(hù)傘在頸動(dòng)脈狹窄支架成形術(shù)中的應(yīng)用[J];第二軍醫(yī)大學(xué)學(xué)報(bào);2002年12期

2 位偉;辛伐他汀對(duì)頸動(dòng)脈粥樣斑塊的影響[J];臨床醫(yī)學(xué);2004年01期

3 孫強(qiáng);韓翠紅;孫學(xué)軍;;缺血預(yù)適應(yīng)和后適應(yīng)的觸發(fā)因子研究進(jìn)展[J];海軍醫(yī)學(xué)雜志;2008年02期

4 羅海蕓;李樹清;;樹

本文編號(hào)：1629088