Abstract
Background Exosomes are small membranous structures released from cells into the blood, regulating various biological processes. However, the role of exosomes in cardiotoxicity remains largely unclear. This study investigated the functional mechanism of exosomal microRNA-96 (miR-96) derived from bone marrow mesenchymal stem cells (BMSCs) in myocardial toxicity induced by doxorubicin. Methods and Results BMSCs were transfected with miR-96 mimic, miR-96 inhibitor, or the negative control before exosome isolation. The functional mechanism of BMSC-derived exosomal miR-96 was investigated in doxorubicin-induced cell and rat models. The cardiac function, histological morphology, and fiber content of myocardium were examined. The expression levels of the following biomarkers were measured for assessment of cardiac injury: creatine kinase isoenzyme MB, cardiac troponin I, brain natriuretic peptide, soluble suppression of tumorigenesis-2, tumor necrosis factor-α, interleukin-1β, interleukin-6, superoxide dismutase, glutathione peroxidase, and malondialdehyde. Cell Counting Kit-8 assay was used to measure the survival rate of cardiomyocytes. The expressions of miR-96, Rac1, p-IKKα/IKKα, p-IKKβ/IKKβ, p-IκBα/IκBα and p-p65/p65 in myocardium and cardiomyocytes were also assessed. The targeting relationship between miR-96 and Rac1 was verified by dual-luciferase reporter assay. miR-96 was downregulated, Rac1 was upregulated and the nuclear factor-κB signaling pathway was activated in doxorubicin-induced cell and animal models. Doxorubicin decreased antioxidative enzymes (superoxide dismutase and glutathione peroxidase) and increased myocardial injury biomarkers (creatine kinase isoenzyme MB, cardiac troponin I, and brain natriuretic peptide), proinflammatory cytokines (tumor necrosis factor-α, interleukin-1β, and interleukin-6), malondialdehyde, and myocardial fibers. Exosomes derived from BMSCs ameliorated doxorubicin-induced myocardial injuries. Overexpression of miR-96 in exosomes derived from BMSCs further enhanced the protection of myocardium and cardiomyocytes against doxorubicin-induced toxicity while miR-96 knockdown abolished the protective effects of exosomes derived from BMSCs. Rac1 was a target gene of miR-96. Rac1 inhibition could downregulate the expression of the nuclear factor-κB signaling and further reverse the promotion of miR-96 knockdown on doxorubicin-induced myocardial toxicity. Conclusions BMSC-derived exosomal miR-96 protects myocardium against doxorubicin-induced toxicity by inhibiting the Rac/nuclear factor-κB signaling pathway.