肾缺血再灌注损伤差异表达长非编码 RNA 分析。

Analysis of Differentially Expressed Long Noncoding RNA in Renal Ischemia-Reperfusion Injury.

机构信息

Department of Critical Care Medicine, The First Affiliated Hospital of Nanchang University, Nanchang, China.

Department of Clinical Laboratory, Affiliated Stomatological Hospital of Nanchang University, Nanchang, China.

出版信息

Kidney Blood Press Res. 2020;45(5):686-701. doi: 10.1159/000508217. Epub 2020 Aug 14.

DOI:10.1159/000508217

PMID:32799207

Abstract

BACKGROUND

Renal ischemia-reperfusion (IR) injury is one of the major causes of acute renal failure which seriously endangers the health and life of patients. Currently, there is still lack of comprehensive knowledge of the molecular mechanism of renal IR injury, and the regulatory role of long noncoding RNA (lncRNA) in renal IR damage remains poorly understood.

AIM

The aim of this study was to analyze the expression spectrum of lncRNA in renal IR damage in mice and to explore specific lncRNA that may be involved in regulating the development of human renal IR injury.

METHODS

RNA-Seq was used to investigate the lncRNA profile of renal IR injury in a mouse model, and conservation analysis was performed on mouse lncRNAs with differential expression (fragments per kilobase of transcript per million mapped reads ≥2) by BLASTN. The potential functions and associated pathways of the differentially expressed lncRNA were explored by bioinformatics analysis. The cell hypoxia model was used to detect the expression of the candidate lncRNA.

RESULTS

Of the 45,923 lncRNA transcripts detected in the samples, and 5,868 lncRNAs were found to be significantly differentially expressed (p < 0.05 and fold change ≥ 2) in 24-h IR kidney tissue compared to the expression in the control group. It was found that 56 differently expressed mouse lncRNA transcripts have human homology by analyzing the conserved sequences. We also found that lncRNA-NONHSAT183385.1 expression significantly increased in HK2 cells after 24 h of hypoxia and increased further 6 h after reoxygenation, and after 24 h of reoxygenation it was dramatically downregulated, indicating that NONHSAT183385.1 may be involved in the pathophysiological process of renal tubular epithelial cells in response to ischemia in human renal IR.

CONCLUSION

Our study revealed differentially expressed lncRNAs in renal IR damage in mice and identified a set of conserved lncRNAs, which would help to explore lncRNAs that may play important regulatory roles in human renal IR injury.

摘要

背景

肾缺血再灌注（IR）损伤是急性肾衰竭的主要原因之一，严重危害患者的健康和生命。目前，对肾 IR 损伤的分子机制仍缺乏全面认识，长链非编码 RNA（lncRNA）在肾 IR 损伤中的调控作用也知之甚少。

目的

本研究旨在分析小鼠肾 IR 损伤中 lncRNA 的表达谱，并探讨可能参与调控人类肾 IR 损伤发生的特定 lncRNA。

方法

采用 RNA-Seq 技术检测小鼠肾 IR 损伤模型中 lncRNA 的表达谱，对差异表达（片段数每千碱基转录本每百万映射reads≥2）的小鼠 lncRNA 进行 BLASTN 保守分析。通过生物信息学分析探讨差异表达 lncRNA 的潜在功能及相关通路。采用细胞低氧模型检测候选 lncRNA 的表达。

结果

在检测到的 45923 个 lncRNA 转录本中，与对照组相比，24 h IR 肾组织中发现 5868 个 lncRNA 表达显著差异（p<0.05，倍数变化≥2）。通过分析保守序列发现，56 个差异表达的小鼠 lncRNA 转录本在人类中具有同源性。我们还发现，在低氧 24 h 后 HK2 细胞中 lncRNA-NONHSAT183385.1 的表达明显增加，再氧合 6 h 后进一步增加，再氧合 24 h 后则明显下调，提示 NONHSAT183385.1 可能参与人肾 IR 肾小管上皮细胞对缺血的病理生理过程。