MiR-10b-5p regulates venous endothelial cells in deep venous thrombosis by targeting MFG-E8

Abstract

IntroductionDeep vein thrombosis (DVT) is a common venous thromboembolic disease with an unclear pathogenesis. miR-10b-5p expression was increased in DVT patients and this study aims to determine the functional mechanism of miR-10b-5p in DVT development.Material and methodsRats were injected with miR-10b-5p and MFG-E8 related plasmids before DVT model establishment. The thrombus length, weight and size were measured. TUNEL staining was used to detect apoptosis of vascular endothelial cells. Cell transfection on HUVECs was performed to achieve miR-10b-5p suppression or/and MFG-E8 suppression. The cell viability, migration, angiogenesis ability and cell apoptosis of HUVECs were measured. The expression levels of miR-10b-5p, MFG-E8, JAK/STAT signal pathway related proteins in peripheral blood of DVT patients, rats, and HUVECs were detected by RT-qPCR and Western blot.ResultsDVT patients increased expression of miR-10b-5p and decreased expression of MFG-E8. miR-10b-5p expression was negatively correlated with MFG-E8. Rats with inhibition of miR-10b-5p or overexpression of MFG-E8 had less deteriorated thrombus, decreased thrombus length and weight, cell apoptosis and suppressed ratio of p-JAK/JAK and p-STAT3/STAT3. miR-10b-5p can negatively regulate and target MFG-E8 in HUVECs. HUVECs transfected with miR-10b-5p inhibitor had increased cell viability, migration and angiogenesis ability as well as suppressed cell apoptosis and decreased p-JAK/JAK and p-STAT3/STAT3 ratio, while co-transfection of miR-10b-5p inhibitor and MFG-E8 suppression can partially counteract the miR-10b-5p inhibitor.ConclusionsElevated expression of miR-10b-5p and decreased MFG-E8 expression were found in peripheral blood of DVT patient and inhibition on miR-10b-5p or overexpression of MFG-E8 can attenuate DVT in rat models.