MicroRNA-4284在下肢动脉硬化闭塞症中的表达特点和意义分析

发布时间：2018-02-12 10:48

  本文关键词： miR-4284 动脉硬化闭塞　出处：《西南医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨微小RNA-4284(MicroRNA-4284简称miR-4284)在下肢动脉硬化闭塞症(Arteriosclerosis obliterans,ASO)患者血浆和病变血管中的表达量变化特点;探讨在缺血清缺氧状态下miR-4284在动脉平滑肌细胞增殖、迁移及凋亡中的作用。方法:临床试验:收集西南医科大学第一附属医院血管甲状腺外科2014-2015年间收治的55例下肢ASO患者和54例健康对照者血浆,24例下肢ASO截肢患者的病变血管及24例外伤截肢患者正常血管。选取其中3对年龄及性别相近的下肢ASO患者和正常健康人血浆用microarray筛选出下肢ASO患者血浆与正常健康人血浆中相比较表达差异在2倍以上的24种MicroRNAs(mi RNAs),用实时荧光定量PCR检测临床55例下肢ASO患者和54例健康对照人群血浆中miR-4284的表达量,24例下肢ASO截肢患者病变血管和24例正常血管中miR-4284的表达量;按Fontaine分期将55例下肢ASO患者分为Ⅰ、Ⅱ、Ⅲ、Ⅳ期,用RT-qPCR分别检测各期患者血浆中mi R-4284的表达量。细胞实验:正常培养主动脉平滑肌、脐静脉内皮细胞后,再进行缺血清缺氧处理用以模拟临床ASO患者下肢缺血缺氧的状态。用高糖培养基培养脐静脉内皮细胞再进行缺血清缺氧处理,用以模拟临床ASO合并Ⅱ型糖尿病患者。采用RT-qPCR检测各组细胞中mi R-4284的相对表达量;分别上调、下调miR-4284后通过cck8检测细胞增殖、transwell检测细胞迁移,流式检测细胞凋亡,蛋白印迹检测凋亡相关蛋白XIAP、BCL2、BAX的表达。结果:临床试验显示,与健康对照组相比下肢ASO患者血浆中miR-4284的表达量在Fontaine第Ⅰ期中就开始显著升高,并在第Ⅱ、Ⅲ、Ⅳ期血浆中一直持续在高水平状态;下肢ASO患者病变血管与正常血管相比,病变血管中miR-4284的表达量也明显升高;细胞试验:脐静脉内皮细胞在低糖及高糖缺血清缺氧的条件下miR-4284的表达量均升高;主动脉平滑肌细胞在缺血清缺氧的条件下miR-4284表达量降低;主动脉平滑肌细胞在缺血清缺氧的条件下,上调、下调miR-4284,对细胞的增殖、迁移没有太大影响;在不进行缺血清缺氧处理的主动脉平滑肌细胞中,上调、下调miR-4284对细胞凋亡没有太大影响;在缺血清缺氧的主动脉平滑肌细胞中上调miR-4284的表达对细胞的凋亡没有太大影响,但下调miR-4284的表达能抑制主动脉平滑肌细胞凋亡;抑制凋亡相关蛋白XIAP、BCL2在下调mi R-4284后表达量明显升高,促凋亡相关蛋白BAX在下调miR-4284后表达量明显降低,差异有统计学意义。结论:miR-4284在下肢ASO患者血浆和血管组织中表达量明显升高;缺血清缺氧诱导主动脉平滑肌细胞中miR-4284表达明显降低,缺血清缺氧诱导人脐静脉内皮细胞中miR-4284表达明显升高;miR-4284对平滑肌细胞增殖、迁移作用不大;下调miR-4284可以抑制平滑肌细胞凋亡,调控miR-4284有望成为靶向治疗下肢ASO研究的新方向。
[Abstract]:Objective: to investigate the changes of micro#en0# expression in plasma and diseased vessels of patients with Arteriosclerosis obliterans obliterans obliterans (Arteriosclerosis obliterans), and to investigate the proliferation of miR-4284 in arterial smooth muscle cells under ischemia and hypoxia. Role of migration and apoptosis. Methods: clinical trial: 55 patients with lower extremity ASO and 54 healthy controls who were treated in vascular thyroid surgery, first affiliated hospital of southwest medical university from 2014 to 2015, 24 patients with lower extremity ASO amputation. Three pairs of patients with age and sex of lower extremity ASO patients and healthy people were selected to use microarray to screen the plasma of lower extremity ASO patients compared with healthy people. Compared with 24 species of MicroRNAs(mi RNAsN, the expression of miR-4284 was detected by real-time fluorescence quantitative PCR in 55 patients with lower extremity ASO and 54 healthy controls. There were 24 patients with ASO amputation of lower extremity and 24 patients with normal blood. The expression of miR-4284 in tube; 55 patients with lower extremity ASO were divided into 鈪，

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