MicroRNA Expression Profile in the Patient's Plasma Exosomes of Alcohol-Induced Osteonecrosis of Femoral Head: Potential Vascular Regulation Mechanism

The purpose of this study is to reveal the expression profile of microRNA (miRNA) in the plasma exosomes of patients with alcohol-induced osteonecrosis of the femoral head (AIONFH), and to further explore the potential effect of alcohol-stimulated hepatocyte exosomes on vascular endothelial cells (VECs). The total RNA of plasma exosomes of normal people (NC-Exos) and patients with AIONFH (AI-Exos) were extracted, followed by Illumina high-throughput sequencing. GO and KEGG analyses were performed. MiR-155-5p in alcohol-stimulated hepatocyte exosomes (AML-Exos) was detected by RT-PCR. The effects of AI-Exos/AML-Exos on the migration, proliferation, and apoptosis of VECs were detected by wound healing assay, CCK-8 assay, and flow cytometry, respectively. The expression of HIF-1 alpha in VECs, a target gene of miR-155-5p, was detected by Western Blot and qRT-PCR. High-throughput sequencing showed that 82 miRNAs were differentially expressed in AI-Exos. RT-PCR results indicated that miR-155-5p, which targets to regulate HIF-1 alpha as confirm via dual-luciferase assay, was the most significantly up-regulated in AI-Exos. Further studies demonstrated that miR-155-5p was specifically up-regulated in AML-Exos. Both AI-Exos and AML-Exos inhibited the proliferation and migration of VECs, promoted their apoptosis, and down-regulated the expression of HIF-1 alpha and VEGFA. This study is the first to reveal the miRNAs expression profile in plasma exosomes of AIONFH patients and suggest a potential mechanism by which exosomes released from alcohol-stimulated hepatocytes may regulate VECs in AIONFH.