Exosomes isolated from the miR-215-modified bone marrow mesenchymal stem cells protect H2O2-induced rat myoblasts via the miR-215/FABP3 pathway

Abstract

This study aimed to investigate the potential effects of miR-215, with exosomes as carriers, against skeletal muscle injury. Exosomes were isolated from rat bone marrow mesenchymal stem cells (rBMSCs) or rBMSCs overexpressing miR-215. Subsequently, rat myoblasts (L6) were treated with different exosomes and mimics, then exposed to H2O2. Cell viability and apoptosis were determined using the Cell Counting Kit-8 and Annexin V-FITC cell apoptosis assay kits, respectively. Reverse-transcriptase quantitative PCR (RT-qPCR) was used to examine the expression of related genes. Transmission electron microscopy, Nanosight, and western blotting showed that the exosomes were successfully isolated. PKH67 staining revealed that both exosomes and miR-215-modified exosomes were taken up by L6 cells. FABP3 was found to be the target gene of miR-215 via a dual luciferase reporter gene assay. In the L6 cells treated with H2O2, cell viability was significantly inhibited, whereas apoptosis significantly increased (P < 0.05). Exosomes significantly enhanced the viability of H2O2-induced cells and inhibited their apoptosis (P < 0.05). In addition, RT-qPCR showed that in the H2O2-induced L6 cells, FABP3, CDKN1A, and TP53 were significantly upregulated, while CCNB1 was significantly downregulated (P < 0.05). However, their expression levels were significantly reversed after treatment with miR-215-modified exosomes (P < 0.05). These findings indicate that the miR-215-modified exosomes may exert protective effects against skeletal muscle injury through the miR-215/FABP3 pathway and regulate the expression of CDKN1A, CCNB1, and TP53.