Transcriptional signature of hypoxic preconditioning in a mouse model of ischemic acute kidney injury

We have performed NGS-derived transcriptome profiling (RNA-seq) to examine the impact of hypoxic preconditioning in post-ischemic kidney injury. Experimental set up: 4 C57BL/6J (male, 8 wks of age) mice were subjected to hypoxia (8%O2) for 48hrs and then subjected to unilateral renal ischemic reperfusion mice (IRI). 4 normoxic littermates subjected to renal IRI served as controls. Injured kidneys were harvested at day 3 following renal IRI and were subjected to NGS. Methods: Poly(A) RNA sequencing library was prepared following Illumina's TruSeq-stranded-mRNA sample preparation protocol. Quality control analysis and quantification of the sequencing library were performed using Agilent Technologies 2100 Bioanalyzer High Sensitivity DNA Chip. Paired-ended sequencing was performed on Illumina's NovaSeq 6000 sequencing system. Overall design: Kidney mRNA profiles at day 3 post ischemic AKI in mice subjected to hypoxic preconditioning compared to normoxia

本研究采用基于下一代测序（Next-Generation Sequencing, NGS）的转录组测序（RNA-seq）技术，探究缺氧预处理对缺血后肾损伤的影响。
实验设置：选取4只8周龄雄性C57BL/6J小鼠，先经8%氧浓度缺氧处理48小时，随后实施单侧肾缺血再灌注（Ischemic Reperfusion Injury, IRI）造模；另取4只同窝仔鼠行常氧处理后进行肾IRI造模，作为对照组。于肾IRI术后第3天采集损伤肾脏，进行NGS测序分析。
实验方法：按照Illumina TruSeq链特异性mRNA样本制备流程构建Poly(A) RNA测序文库；采用安捷伦（Agilent Technologies）2100生物分析仪高灵敏度DNA芯片完成测序文库的质控与定量；使用Illumina NovaSeq 6000测序系统开展双端测序。
整体实验设计：对比缺氧预处理组与常氧组小鼠缺血性急性肾损伤（Acute Kidney Injury, AKI）术后第3天的肾脏mRNA表达谱。