High-throughput Sequencing contributes to Quantitative Analysis of Renal Ischemic-Reperfusion-Injury Transcriptomes

Purpose: The different ischemic time of Ischemia-reperfusion-injury (IRI) mouse models resulted in different outcomes, severe IRI led to chronic kidney disease, while mild IRI led to the recovery of kidneys. We isolated tubules from mice from each group. The goals of this study are to compare differentially expressed mRNAs, lncRNAs and circRNAs among severe IRI-injured tubules, mild IRI-injuried tubules and control mouse tubules. Methods: kidney tubular mRNA, lncRNA, circRNA profiles of severe IRI (40-min ischemia), mild IRI (20-min ischemia) and control mice were generated by deep sequencing, in triplicate, using Illumina HiSeq 4000. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using SYBR Green assays.