本文關(guān)鍵詞： 顱內(nèi)動(dòng)脈瘤 微小RNA-b 平滑肌細(xì)胞 自噬基因 細(xì)胞自噬　出處：《山東醫(yī)藥》2017年05期 　論文類型：期刊論文

【摘要】：目的觀察微小RNA-29b(miRNA-29b)在顱內(nèi)動(dòng)脈瘤組織中的表達(dá)變化,并探討其意義。方法取20例顱內(nèi)動(dòng)脈瘤患者顱內(nèi)動(dòng)脈瘤組織、腦腫瘤患者正常腦動(dòng)脈作為標(biāo)本;制備大鼠顱內(nèi)動(dòng)脈瘤模型,取其顱內(nèi)動(dòng)脈瘤組織及正常腦動(dòng)脈作為標(biāo)本。用qRT-PCR技術(shù)檢測(cè)以上標(biāo)本中miRNA-29b表達(dá)。將培養(yǎng)好的人主動(dòng)脈平滑肌細(xì)胞隨機(jī)分為實(shí)驗(yàn)組和對(duì)照組,按照Lipofectamine 2000將miRNA-29b mimics、miRNA mimics NC分別轉(zhuǎn)染入實(shí)驗(yàn)組、對(duì)照組,轉(zhuǎn)染24 h取實(shí)驗(yàn)組部分細(xì)胞經(jīng)20 mg/L ox-LDL干預(yù)24 h。用qRT-PCR技術(shù)檢測(cè)細(xì)胞內(nèi)miRNA-29b、自噬基因(Beclin1)mRNA表達(dá);用Western blotting法檢測(cè)細(xì)胞內(nèi)Beclin1、微管相關(guān)蛋白(LC3Ⅱ/Ⅰ)蛋白表達(dá);用雙熒光素酶實(shí)驗(yàn)檢驗(yàn)Beclin1是否為miRNA-29b的靶基因;用MTT法檢測(cè)細(xì)胞增殖率。結(jié)果人與大鼠顱內(nèi)動(dòng)脈瘤組織中miRNA-29b相對(duì)表達(dá)量均低于正常腦動(dòng)脈組織(P均0.05)。經(jīng)ox-LDL刺激的人平滑肌細(xì)胞內(nèi)miRNA-29b相對(duì)表達(dá)量低于未經(jīng)ox-LDL刺激的細(xì)胞(P0.05)。轉(zhuǎn)染48 h,實(shí)驗(yàn)組Beclin1 mRNA及蛋白相對(duì)表量低于對(duì)照組,LC3Ⅱ/Ⅰ蛋白相對(duì)表達(dá)量高于對(duì)照組(P均0.05)。熒光素酶實(shí)驗(yàn)證實(shí)miRNA-29b可與Beclin1 mRNA的3'-UTR結(jié)合,Beclin1是miRNA-29b的靶基因。轉(zhuǎn)染48 h,實(shí)驗(yàn)組細(xì)胞增殖率低于對(duì)照組(P0.05)。結(jié)論 miRNA-29b在顱內(nèi)動(dòng)脈瘤組織中表達(dá)下調(diào),其可能通過介導(dǎo)靶基因Beclin1調(diào)控平滑肌細(xì)胞的自噬,從而參與顱內(nèi)動(dòng)脈瘤的發(fā)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，

本文編號(hào)：1505231