Department of Nephrology, First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, People's Republic of China.
Department of Urology, First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, People's Republic of China.
Gene. 2018 Jul 20;664:127-138. doi: 10.1016/j.gene.2018.04.028. Epub 2018 Apr 12.
To analyze and preliminarily validate the lncRNA expression profiles in the stenosis tissue of arteriovenous fistula (AVF).
A total of 54 uremic patients administered in the department of nephrology in the First Affiliated Hospital of Nanchang University between February 2017 and March 2017 were included in the study and subsequently categorized as experimental group, which included 12 patients with confirmed diagnosis of AVF, and control group which included 42 patients with no vascular stenosis. The experimental group inclusion criteria include: AVF used >3 months; The blood flow of AVF <200 mL/min; the degree of the stenosis was >50%, excluded obvious thrombosis; The ultrasonographic data of AVF stenosis is complete. The controls were randomly selected from uremic patients who were primary AVF operation, excluded obvious vascular stenosis and vascular diseases. Among them, 4 sample in the experimental group and 4 controls were used in LncRNA sequencing. RNA in vascular tissue was extracted by Trizol and IncRNA sequencing was used to establish the expression profiles of lncRNA in the stenosis tissue of AVF.9 difference expression lncRNA were collected for validating in AVF stenosis cases by using quantitative real-time polymerase chain reaction (qRT-PCR). Moreover, Cluster analysis, gene functional analysis and pathway analysis were used to explore the function of difference expression lncRNA.
Among the 27,692 lncRNA transcripts examined, 247 lncRNAs were found to be significantly differentially expressed (P < 0.05, fold change ≥2) in the experimental group and control group, with 141 being up-regulated and 106 down-regulated. The expression levels of 9 lncRNAs validated by subsequent qRT-PCR were shown to be highly consistent with the sequencing data.
Our study revealed lncRNAs expression profiles in the stenosis tissue of AVF by LncRNA sequencing. These lncRNAs and its related signaling pathways may play a key role in the occurrence and progression of AVF stenosis.
分析并初步验证动静脉瘘（AVF）狭窄组织中的 lncRNA 表达谱。
选取 2017 年 2 月至 2017 年 3 月南昌大学第一附属医院肾内科收治的 54 例尿毒症患者，分为实验组和对照组。实验组 12 例患者均确诊为 AVF，对照组 42 例患者无血管狭窄。实验组纳入标准：AVF 使用>3 个月；AVF 血流量<200ml/min；狭窄程度>50%，无明显血栓形成；AVF 狭窄的超声数据完整。对照组从原发性 AVF 手术的尿毒症患者中随机选择，排除明显血管狭窄和血管疾病。其中，实验组 4 例样本和对照组 4 例用于 lncRNA 测序。采用 Trizol 提取血管组织中的 RNA，建立 AVF 狭窄组织中 lncRNA 的表达谱。采用定量实时聚合酶链反应（qRT-PCR）对 9 个差异表达 lncRNA 在 AVF 狭窄病例中的表达进行验证。此外，还进行了聚类分析、基因功能分析和通路分析，以探讨差异表达 lncRNA 的功能。
在检测的 27692 个 lncRNA 转录本中，实验组和对照组有 247 个 lncRNA 表达显著差异（P<0.05，倍数变化≥2），其中 141 个上调，106 个下调。后续 qRT-PCR 验证的 9 个 lncRNA 的表达水平与测序数据高度一致。
本研究通过 LncRNA 测序揭示了 AVF 狭窄组织中的 lncRNA 表达谱。这些 lncRNA 及其相关信号通路可能在 AVF 狭窄的发生和进展中发挥关键作用。
