The changes of miRNAs in plasma small extracellular vesicles (sEV) isolated from 8-week diabetic rats compared with control animals
Compared with plasma sEV (Con-sEV) from control rats, plasma sEV (DM-sEV) from 8-week diabetic rats significantly induced cardiomyocyte apoptosis as evidenced by increased percentage of apoptotic cells and activity of pro-apoptotic protein caspase 3. The proapoptotic effect of DM-sEV was blunted by RNase but not proteinase K, suggesting that DM-sEV exerted the cardiotoxic effects mainly through their contained RNAs. Increasing evidence suggests that miRNAs are the most important molecules by which sEV regulate recipient cell function. To identify  the sEV-containing specific miRNAs responsible for the effects, Con-sEV and DM-sEV were subjected to miRNA sequencing. Overall design: To explore the changes of miRNAs in plasma DM-sEV compared with Con-sEV, Con-sEV and DM-sEV were isolated from the plasma of control and 8-week diabetic rats. Total RNAs were extracted from sEV purified from 2 ml of rat plasma. The PCR products derived from the 18- to 30- nucleotide RNA molecules were purified by electrophoresis and sequenced using the Illumina HiSeq 2500 platform. Differential expression analysis of two groups was performed using the DEGseq R package.