细胞自噬在紫绀型先心病心肌慢性缺氧适应中的机制的研究

发布时间：2018-06-08 21:00

  本文选题：细胞自噬 + 自噬体　； 参考：《蚌埠医学院》2017年硕士论文

【摘要】：目的:本实验通过对紫绀型先心病患儿心肌组织中细胞自噬现象的检测,进而研究细胞自噬在心肌慢性缺氧适应性中的表达,对细胞自噬在缺氧心肌中的作用进行探讨。为如何于缺氧条件下减少心肌的损伤及改善紫绀型先心病患儿的治疗现况提供理论依据。方法:实验组:蚌埠医学院第一附属医院与南京儿童医院心脏外科2015年4月至2016年10月治疗的20例紫绀型先心病患儿,平均年龄为19.65±8.59月。对照组:收集蚌埠医学院第一附属医院与南京儿童医院心脏外科2015年4月至2016年10月治疗的20例非紫绀型先心病患儿,平均年龄为16.55±7.24月。取术中患儿右室流出道肥厚心肌组织为标本,一部分大小为5×5×5mm3放置于液氮罐中用于Western-blotting检测,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以下的样本制成超薄切片后通过电子透镜观察心肌组织中自噬体的形成情况;提取液氮储存的标本的心肌组织蛋白通过Western-blotting检测心肌组织中LC3Ⅱ/LC3Ⅰ比值的大小和Beclin-1蛋白表达水平,然后对组间相关的数据进行分析。结果:通过电子透镜观察表明实验组和对照组相比,实验组心肌组织内细胞器排列紊乱,线粒体增多,大量的细胞器被降解,内质网肿胀,细胞器结构模糊,可见包绕部分细胞器及蛋白质的自噬小体。对照组细胞器降解较少,肌丝排列规律,无自噬小体的出现。Western-blotting表明试验组紫绀型先心病患儿LC3Ⅱ表达较高,且LC3Ⅱ/LC3Ⅰ比值明显大于对照非紫绀组,Beclin-1蛋白表达水平紫绀组明显高于非紫绀组。用SPSS18.0软件行t检验,结果显示两组之间LC3和Beclin-1表达水平的存有组间差异(p0.05),表明两组之间对比表现的差异具有统计学意义,实验证明了在缺氧条件下心肌组织自噬现象明显被激活。结论:1.紫绀型先心病患儿心肌组织中电镜发现自噬体的形成,表明缺氧心肌细胞中存在细胞自噬的现象;2.紫绀组患儿LC3Ⅱ表达较高,且LC3Ⅱ/LC3Ⅰ比值明显大于对照组,Beclin-1蛋白表达水平实验组明显高于对照组。3.细胞自噬在心肌慢性缺氧中影响心肌的存活,在紫绀型先心病发生发展中起调控作用。
[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 卤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 卤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，

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