本文選題：顱內(nèi)動(dòng)脈瘤 + 血管平滑肌細(xì)胞��； 參考：《蘇州大學(xué)》2016年碩士論文

【摘要】：目的:顱內(nèi)動(dòng)脈瘤(intracranial aneurysm,IA)具有自發(fā)性破裂出血的傾向,致殘率和致死率都很高。目前,在臨床上關(guān)于IA的治療僅僅局限于手術(shù)和介入治療,而對(duì)動(dòng)脈瘤壁進(jìn)行修復(fù)等非手術(shù)治療幾乎沒有。在對(duì)IA發(fā)病機(jī)制的研究中,我們發(fā)現(xiàn)動(dòng)脈瘤壁血管平滑肌細(xì)胞(vascular smooth muscle cell,VSMC)具有增殖活性功能,結(jié)合目前生物治療和基因治療的發(fā)展,我們萌生了為未破裂顱內(nèi)動(dòng)脈瘤壁進(jìn)行修復(fù)治療的設(shè)想。VSMC是動(dòng)脈壁組成的主要部分,在IA形成過程中,可表現(xiàn)出很高的增殖活性與可塑性,可見IA形成過程中血管壁VSMC的增殖將成為修復(fù)研究的主要方向。因此,本實(shí)驗(yàn)通過建立大鼠顱內(nèi)動(dòng)脈瘤模型對(duì)大鼠顱內(nèi)動(dòng)脈瘤壁VSMC增殖活性進(jìn)行探討,目的在于增強(qiáng)動(dòng)脈瘤壁VSMC的增殖活性,增加平滑肌層,為今后人顱內(nèi)動(dòng)脈瘤壁的修復(fù)治療提供理論基礎(chǔ)。方法:成年雄性自發(fā)性高血壓大鼠(spontaneously hypertensive rat,SHR)180只,隨機(jī)分為6組:對(duì)照組和實(shí)驗(yàn)1、2、3、4、5月組(每組30只)。實(shí)驗(yàn)組采用經(jīng)背部入路行雙側(cè)腎動(dòng)脈后支結(jié)扎和左側(cè)頸總動(dòng)脈結(jié)扎,對(duì)照組僅暴露血管,不結(jié)扎。各組術(shù)后1周開始喂養(yǎng)含1%氯化鈉的飲用水和0.12%β-氨基丙腈飼料。每組大鼠術(shù)前及處死前測(cè)量室溫安靜狀態(tài)下尾動(dòng)脈血壓,處死前行7.0MRA檢查大鼠顱內(nèi)動(dòng)脈。取下動(dòng)脈瘤標(biāo)本,用HE染色觀察動(dòng)脈瘤的病理變化,免疫組化觀察瘤壁增殖細(xì)胞核抗原(proliferating cell nuclear antigen,PCNA)的表達(dá)和免疫熒光染色和蛋白質(zhì)印跡(Westernblot,WB)觀察瘤壁α-平滑肌肌動(dòng)蛋白(α-smooth muscle actin,α-SMA)、β-微管蛋白(β-Tubulin)和骨橋蛋白(osteopontin,OPN)的表達(dá)以及實(shí)時(shí)定量PCR觀測(cè)瘤壁SM22α與高血壓相關(guān)基因(hypertension-related gene,HRG-1)mRNA的表達(dá)來了解動(dòng)脈瘤壁VSMC的增殖,末端標(biāo)記法(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling,TUNEL)染色測(cè)定瘤壁VSMC的凋亡。結(jié)果:1.血壓變化:對(duì)照組平均血壓緩慢升高。實(shí)驗(yàn)組平均血壓從1月組起至5月組逐漸升高,均明顯高于術(shù)前和對(duì)照組(P0.05)。2.MRA檢測(cè)情況:每組隨機(jī)抽取6只SHR行7.0 MRA檢測(cè)。對(duì)照組顱內(nèi)動(dòng)脈左右對(duì)稱、粗細(xì)均勻,無明顯擴(kuò)張?jiān)龃帧?shí)驗(yàn)組所有檢測(cè)的右側(cè)顱內(nèi)動(dòng)脈均有不同程度的增粗,血管病變率達(dá)100%,其中,4月組有3/6在增粗的血管上可見動(dòng)脈瘤形成,5月組有5/6可見動(dòng)脈瘤形成。3.動(dòng)脈瘤病理變化:對(duì)照組顱內(nèi)動(dòng)脈無明顯破壞,各層結(jié)構(gòu)保持完整。實(shí)驗(yàn)組顱內(nèi)動(dòng)脈從1月組起內(nèi)膜層開始破壞、VSMC退變、VSMC數(shù)目及層數(shù)減少、動(dòng)脈壁逐漸變薄,至4、5月組時(shí)內(nèi)膜層已消失、彈性纖維斷裂,并有炎性細(xì)胞浸潤(rùn),其中,5月組最明顯,動(dòng)脈壁最薄。4.免疫組化結(jié)果:對(duì)照組PCNA陽性細(xì)胞率低下。實(shí)驗(yàn)組PCNA陽性細(xì)胞率從1月組起至5月組與對(duì)照組相比均明顯增高(P0.05);其中,4月組最高,5月組開始下降,與4月組相比有顯著差異(P0.05)。5.Western blot結(jié)果:對(duì)照組α-SMA和β-tubulin呈高表達(dá),而OPN幾乎不表達(dá)。實(shí)驗(yàn)組α-SMA和β-tubulin的表達(dá)從1月組起至5月組逐漸下降,均明顯低于對(duì)照組(P0.05);而OPN的表達(dá)從1月組起至5月組與對(duì)照組相比均明顯增高(P0.05);其中,4月組最高,5月組開始下降,與4月組相比差異顯著(P0.05)。6.免疫熒光結(jié)果:對(duì)照組α-SMA和β-tubulin呈高表達(dá),而OPN幾乎不表達(dá)。實(shí)驗(yàn)組α-SMA和β-Tubulin的相對(duì)熒光強(qiáng)度從1月組起至5月組逐漸降低,均明顯低于對(duì)照組(P0.05);而OPN的相對(duì)熒光強(qiáng)度從1月組起至5月組與對(duì)照組相比均明顯增高(P0.05),其中,4月組最高,5月組開始下降,與4月組相比差異顯著(P0.05)。7.實(shí)時(shí)定量PCR結(jié)果:對(duì)照組SM22α和HRG-1 mRNA相對(duì)表達(dá)較高。實(shí)驗(yàn)組SM22α和HRG-1 mRNA相對(duì)表達(dá)從1月組起至5月組逐漸下降,均明顯低于對(duì)照組(P0.05)。8.TUNEL染色結(jié)果:對(duì)照組VSMC凋亡率低下。實(shí)驗(yàn)組VSMC凋亡率從1月組起至5月組逐漸升高,均明顯高于對(duì)照組(P0.05)。結(jié)論:實(shí)驗(yàn)性大鼠IA形成過程中動(dòng)脈瘤壁VSMC具有增殖活性功能,且具有明顯的增殖活躍期;IA的形成與動(dòng)脈瘤壁VSMC的增殖活性相對(duì)減弱密切相關(guān)。
[Abstract]:Objective: intracranial aneurysm (IA) has a tendency to spontaneous rupture and hemorrhage, and the rate of disability and lethality are high. Currently, the clinical treatment of IA is limited to surgery and interventional therapy, but nonsurgical treatment for the repair of the aneurysm wall is few. In the study of the pathogenesis of IA, we found the movement. Vascular smooth muscle cell (VSMC) has the function of proliferative activity. Combined with the development of biological therapy and gene therapy, we conceive that.VSMC is the main part of the arterial wall composition, which can show high proliferation in the process of IA formation. The proliferation of the vascular wall VSMC in the formation of IA will be the main direction of the repair research. Therefore, the rat intracranial aneurysm model was established to explore the VSMC proliferation activity of the intracranial aneurysm wall in rats. The aim of this experiment is to enhance the proliferation activity of the aneurysm wall VSMC and increase the smooth muscle layer for the future human intracranial. The repair of aneurysm wall provides a theoretical basis. Methods: 180 adult male spontaneously hypertensive rats (spontaneously hypertensive rat, SHR) were randomly divided into 6 groups: the control group and the experimental 1,2,3,4,5 month group (30 rats in each group). The experimental group was ligation of the posterior branches of the bilateral renal artery and the left common carotid artery by the back approach, and the control group was only violent. 1 weeks after operation, 1% sodium chloride containing drinking water and 0.12% beta aminonitrile feed were fed. The blood pressure of the caudal artery was measured at room temperature and quiet before and before the operation. Before and before the operation, the intracranial arteries of the rats were examined by 7.0MRA. The specimens of the aneurysm were taken and the pathological changes of the aneurysm were observed by HE staining and the immunohistochemical view was observed. The expression of proliferating cell nuclear antigen (PCNA) and immunofluorescence staining and Western blot (Westernblot, WB) observation of the tumor wall alpha smooth muscle actin (alpha -smooth muscle actin, alpha -SMA), the expression of beta microtubulin (beta -Tubulin) and osteopontin and real-time quantitative observation The tumor wall SM22 alpha and hypertension-related gene (HRG-1) mRNA were expressed to understand the proliferation of the aneurysm wall VSMC. Terminal labeling (terminal deoxynucleotidyl transferase-mediated dUTP nick end) staining was used to determine the apoptosis of the tumor wall. Results: 1. blood pressure changes: the mean blood pressure in the control group increased slowly. The average blood pressure in the experimental group increased gradually from the January group to the May group, which was significantly higher than that in the pre operation and the control group (P0.05).2.MRA detection: each group was randomly selected 6 SHR for 7 MRA tests. The control group was symmetrical and thin, without obvious dilation and thickening. All of the right intracranial arteries in the experimental group were thickened in varying degrees. The rate of vascular lesions was 100%, of which, in the April group, the aneurysm was visible on the thickened vessels, and the pathological changes of the aneurysm formed in the group of 5/6 were seen in the May group. There was no obvious destruction of the intracranial artery in the control group, and the structure of each layer remained intact. The intracranial artery in the experimental group began to destroy from the inner membrane of the January group, the VSMC degeneration, the number of VSMC and the number of layers decreased in the experimental group. The arterial wall gradually thinned, the intima layer disappeared, the elastic fibers broke and the inflammatory cells infiltrated in the 4,5 month group. Among them, the May group was the most obvious, the thinnest.4. immunohistochemical results were found in the control group, and the rate of PCNA positive cells in the control group was low. The rate of PCNA positive cells in the experimental group increased significantly from the January group to the May group (P0.05), and 4 The month group was the highest, and the May group began to decline. Compared with the April group, there was a significant difference (P0.05).5.Western blot results: the control group of alpha -SMA and beta -tubulin were highly expressed, and OPN almost did not express. The expression of alpha -SMA and beta -tubulin in the experimental group decreased gradually from the January group to the May group, all significantly lower than the control group (P0.05), while the expression of OPN was from January to May group. Compared with the control group (P0.05), the April group was the highest and the May group began to decline. Compared with the April group, the difference was significant (P0.05).6. immunofluorescence results: the control group of alpha -SMA and beta -tubulin were highly expressed, and the OPN almost did not express. The relative fluorescence intensity of the alpha -SMA and beta -Tubulin in the experimental group decreased gradually from the January group to the May group, both significantly lower. Compared with the control group (P0.05), the relative fluorescence intensity of OPN increased significantly from the January group to the May group (P0.05), which was the highest in the April group, and the May group began to decline. Compared with the April group, the difference was significant (P0.05).7. real-time quantitative PCR results: the relative expression of SM22 alpha and HRG-1 mRNA in the control group was higher. The SM22 A and HRG-1 mRNA in the experimental group were relatively expressed. From the January group to the May group gradually decreased, both significantly lower than the control group (P0.05).8.TUNEL staining results: the control group VSMC apoptosis rate is low. The apoptosis rate of VSMC in the experimental group increased gradually from the January group to the May group, all obviously higher than the control group (P0.05). Conclusion: the arterial wall VSMC has the proliferative activity function in the experimental rat IA formation process, and it has a bright future. The proliferative phase was significantly correlated with the formation of IA and the relative attenuation of VSMC proliferation in aneurysm walls.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R743

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2 蒲朝霞;主動(dòng)脈內(nèi)徑和彈性改變與顱內(nèi)動(dòng)脈瘤的相關(guān)性研究[D];浙江大學(xué);2011年

3 沙龍金;炎癥相關(guān)基因多態(tài)性與168例顱內(nèi)動(dòng)脈瘤易感性的關(guān)聯(lián)研究[D];南方醫(yī)科大學(xué);2014年

4 伍剛;血管平滑肌雌激素受體表達(dá)與顱內(nèi)動(dòng)脈瘤相關(guān)性研究[D];中國(guó)人民解放軍軍醫(yī)進(jìn)修學(xué)院;2007年

5 陳文華;顱內(nèi)動(dòng)脈瘤的診斷、治療和療效評(píng)價(jià)的比較影像學(xué)研究[D];南京醫(yī)科大學(xué);2009年

6 葛明旭;顱內(nèi)動(dòng)脈瘤致病基因篩查及血管內(nèi)介入治療的研究[D];山東大學(xué);2007年

7 朱文煥;顱內(nèi)動(dòng)脈瘤動(dòng)物模型的建立及發(fā)病機(jī)制的研究[D];蘇州大學(xué);2012年

8 萬子昂;基于患者特征的顱內(nèi)動(dòng)脈瘤破裂相關(guān)危險(xiǎn)因素研究[D];浙江大學(xué);2015年

9 虞軍;全外顯子組測(cè)序探尋顱內(nèi)動(dòng)脈瘤家系致病基因的研究[D];浙江大學(xué);2013年

10 辛濤;數(shù)字減影三維重建在顱內(nèi)動(dòng)脈瘤影像診斷和介入治療中的應(yīng)用[D];第二軍醫(yī)大學(xué);2005年

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1 索苗苗;1、LncRNA AL110200促進(jìn)動(dòng)脈粥樣硬化炎癥作用及機(jī)制研究 2、Kallikrein基因遺傳變異與國(guó)人散發(fā)性顱內(nèi)動(dòng)脈瘤的關(guān)聯(lián)性研究[D];北京協(xié)和醫(yī)學(xué)院;2013年

2 宋洋;顱內(nèi)動(dòng)脈瘤破裂急性期治療方案的Meta-分析[D];山西醫(yī)科大學(xué);2015年

3 宗華;120例顱內(nèi)動(dòng)脈瘤血管內(nèi)介入治療的臨床研究[D];山西醫(yī)科大學(xué);2015年

4 薛文濤;血管內(nèi)栓塞治療顱內(nèi)動(dòng)脈瘤的臨床資料分析[D];青島大學(xué);2015年

5 馬劉佳;顱內(nèi)動(dòng)脈瘤破裂早期血管內(nèi)栓塞治療臨床分析[D];延安大學(xué);2015年

6 張利通;天津地區(qū)顱內(nèi)動(dòng)脈瘤危險(xiǎn)因素相關(guān)基因多態(tài)性的研究[D];天津醫(yī)科大學(xué);2015年

7 張騰;顱內(nèi)動(dòng)脈瘤破裂的危險(xiǎn)因素分析[D];濟(jì)南大學(xué);2015年

8 何巍;顱內(nèi)動(dòng)脈瘤顯微夾閉術(shù)治療效果病例分析[D];大連醫(yī)科大學(xué);2015年

9 何雪峰;高級(jí)別顱內(nèi)動(dòng)脈瘤患者保守治療12h后再手術(shù)對(duì)預(yù)后的影響[D];山西醫(yī)科大學(xué);2016年

10 康慧斌;顱內(nèi)動(dòng)脈瘤破裂臨床風(fēng)險(xiǎn)因素分析[D];首都醫(yī)科大學(xué);2016年

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