miRNA-29b在颅内动脉瘤组织中的表达变化及意义

发布时间：2018-02-12 08:44

  本文关键词： 颅内动脉瘤 微小RNA-b 平滑肌细胞 自噬基因 细胞自噬　出处：《山东医药》2017年05期 　论文类型：期刊论文

【摘要】：目的观察微小RNA-29b(miRNA-29b)在颅内动脉瘤组织中的表达变化,并探讨其意义。方法取20例颅内动脉瘤患者颅内动脉瘤组织、脑肿瘤患者正常脑动脉作为标本;制备大鼠颅内动脉瘤模型,取其颅内动脉瘤组织及正常脑动脉作为标本。用qRT-PCR技术检测以上标本中miRNA-29b表达。将培养好的人主动脉平滑肌细胞随机分为实验组和对照组,按照Lipofectamine 2000将miRNA-29b mimics、miRNA mimics NC分别转染入实验组、对照组,转染24 h取实验组部分细胞经20 mg/L ox-LDL干预24 h。用qRT-PCR技术检测细胞内miRNA-29b、自噬基因(Beclin1)mRNA表达;用Western blotting法检测细胞内Beclin1、微管相关蛋白(LC3Ⅱ/Ⅰ)蛋白表达;用双荧光素酶实验检验Beclin1是否为miRNA-29b的靶基因;用MTT法检测细胞增殖率。结果人与大鼠颅内动脉瘤组织中miRNA-29b相对表达量均低于正常脑动脉组织(P均0.05)。经ox-LDL刺激的人平滑肌细胞内miRNA-29b相对表达量低于未经ox-LDL刺激的细胞(P0.05)。转染48 h,实验组Beclin1 mRNA及蛋白相对表量低于对照组,LC3Ⅱ/Ⅰ蛋白相对表达量高于对照组(P均0.05)。荧光素酶实验证实miRNA-29b可与Beclin1 mRNA的3'-UTR结合,Beclin1是miRNA-29b的靶基因。转染48 h,实验组细胞增殖率低于对照组(P0.05)。结论 miRNA-29b在颅内动脉瘤组织中表达下调,其可能通过介导靶基因Beclin1调控平滑肌细胞的自噬,从而参与颅内动脉瘤的发生。
[Abstract]:Objective to observe the expression and significance of microRNA-29bmmiRNA-29b in intracranial aneurysms. Methods Intracranial aneurysms were collected from 20 patients with intracranial aneurysms and normal cerebral arteries were collected from 20 patients with intracranial aneurysms. The qRT-PCR technique was used to detect the expression of miRNA-29b in the above specimens. The cultured human aortic smooth muscle cells were randomly divided into experimental group and control group. According to Lipofectamine 2000, the miRNA-29b mimicsm miRNA mimics NC was transfected into the experimental group and the control group respectively. After 24 h transfection, some cells of the experimental group were treated with 20 mg/L ox-LDL for 24 h. The intracellular miRNA-29bmRNA expression of autophagy gene was detected by qRT-PCR technique. Western blotting assay was used to detect the expression of Beclin1, microtubule-associated protein LC3 鈪，

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