Exosomal miRNA and mRNA Sequencing in L-Arabinose-Treated Intestinal Injury Mouse Model
This study aimed to investigate the protective effects of L-arabinose, a naturally occurring pentose sugar, on cyclophosphamide-induced intestinal injury in mice, through integrated analyses of fecal exosomal miRNA sequencing and small intestinal transcriptome sequencing.Intestinal injury was induced in male C57BL/6J mice by intraperitoneal injection of cyclophosphamide (50 mg/kg body weight) for 6 consecutive days. Mice were simultaneously treated with L-arabinose at two doses (200 mg/kg/day and 400 mg/kg/day) via daily gavage.1. Exosomal miRNA sequencing: Fecal samples were collected from mice. Exosomes were isolated by ultracentrifugation (100,000 g for 70 min) and characterized by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). Total RNA was extracted from exosomes using the miRNeasy Mini Kit (Qiagen). Small RNA libraries were constructed with the NEBNext Multiplex Small RNA Library Prep Set for Illumina and sequenced on an Illumina NovaSeq 6000 platform (single-end 50 bp). Raw reads were quality-filtered using cutadapt (v3.5) and Trim Galore (v0.6.7).2. Small intestinal transcriptome sequencing: Small intestinal tissues were collected from the same cohort of mice. Total RNA was extracted and used for transcriptome library construction, followed by high-throughput sequencing on the Illumina platform and quality control.Four biological replicates per group (control, injury model, injury + low-dose L-arabinose, injury + high-dose L-arabinose) were included. This multi-omics study aims to reveal the molecular mechanism by which L-arabinose regulates intestinal injury and repair.