本文選題：細胞自噬 + 自噬體　； 參考：《蚌埠醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:本實驗通過對紫紺型先心病患兒心肌組織中細胞自噬現(xiàn)象的檢測,進而研究細胞自噬在心肌慢性缺氧適應(yīng)性中的表達,對細胞自噬在缺氧心肌中的作用進行探討。為如何于缺氧條件下減少心肌的損傷及改善紫紺型先心病患兒的治療現(xiàn)況提供理論依據(jù)。方法:實驗組:蚌埠醫(yī)學(xué)院第一附屬醫(yī)院與南京兒童醫(yī)院心臟外科2015年4月至2016年10月治療的20例紫紺型先心病患兒,平均年齡為19.65±8.59月。對照組:收集蚌埠醫(yī)學(xué)院第一附屬醫(yī)院與南京兒童醫(yī)院心臟外科2015年4月至2016年10月治療的20例非紫紺型先心病患兒,平均年齡為16.55±7.24月。取術(shù)中患兒右室流出道肥厚心肌組織為標本,一部分大小為5×5×5mm3放置于液氮罐中用于Western-blotting檢測,另一部分快速切成1mm3以下,用5%的戊二醛固定,置于EP管中4℃避光保存。1mm3以下的樣本制成超薄切片后通過電子透鏡觀察心肌組織中自噬體的形成情況;提取液氮儲存的標本的心肌組織蛋白通過Western-blotting檢測心肌組織中LC3Ⅱ/LC3Ⅰ比值的大小和Beclin-1蛋白表達水平,然后對組間相關(guān)的數(shù)據(jù)進行分析。結(jié)果:通過電子透鏡觀察表明實驗組和對照組相比,實驗組心肌組織內(nèi)細胞器排列紊亂,線粒體增多,大量的細胞器被降解,內(nèi)質(zhì)網(wǎng)腫脹,細胞器結(jié)構(gòu)模糊,可見包繞部分細胞器及蛋白質(zhì)的自噬小體。對照組細胞器降解較少,肌絲排列規(guī)律,無自噬小體的出現(xiàn)。Western-blotting表明試驗組紫紺型先心病患兒LC3Ⅱ表達較高,且LC3Ⅱ/LC3Ⅰ比值明顯大于對照非紫紺組,Beclin-1蛋白表達水平紫紺組明顯高于非紫紺組。用SPSS18.0軟件行t檢驗,結(jié)果顯示兩組之間LC3和Beclin-1表達水平的存有組間差異(p0.05),表明兩組之間對比表現(xiàn)的差異具有統(tǒng)計學(xué)意義,實驗證明了在缺氧條件下心肌組織自噬現(xiàn)象明顯被激活。結(jié)論:1.紫紺型先心病患兒心肌組織中電鏡發(fā)現(xiàn)自噬體的形成,表明缺氧心肌細胞中存在細胞自噬的現(xiàn)象;2.紫紺組患兒LC3Ⅱ表達較高,且LC3Ⅱ/LC3Ⅰ比值明顯大于對照組,Beclin-1蛋白表達水平實驗組明顯高于對照組。3.細胞自噬在心肌慢性缺氧中影響心肌的存活,在紫紺型先心病發(fā)生發(fā)展中起調(diào)控作用。
[Abstract]:Aim: to study the expression of autophagy in chronic hypoxic adaptation and to explore the role of autophagy in hypoxic myocardium by detecting the phenomenon of autophagy in myocardial tissue of children with cyanotic congenital heart disease. To provide theoretical basis for reducing myocardial injury and improving the current treatment of cyanotic congenital heart disease in hypoxic condition. Methods: the experimental group consisted of 20 children with cyanotic congenital heart disease treated in the first affiliated Hospital of Bengbu Medical College and Nanjing Children's Hospital from April 2015 to October 2016. The average age was 19.65 鹵8.59 months. Control group: 20 children with non-cyanotic congenital heart disease were collected from the first affiliated Hospital of Bengbu Medical College and the Children's Hospital of Nanjing from April 2015 to October 2016. The average age was 16.55 鹵7.24 months. The right ventricular outflow tract (RVOT) hypertrophic myocardial tissue was taken as a specimen, some of which was 5 脳 5 脳 5mm3 in liquid nitrogen tank for Western-blotting, the other was quickly cut below 1mm3 and fixed with 5% glutaraldehyde. Ultrathin sections were made from samples stored in EP tube at 4 鈩，

本文編號：1997082