MicroRNA-125a-5p(miR-125a-5p)could participate in the pathogenesis of vascular diseases. In this study, we investigated the role of miR-125a-5p in oxidized low density lipoprotein (ox-LDL) induced functional changes in human brain microvessel endothelial cells (HBMEC). The reactive oxygen species (ROS) production, nitric oxide (NO) generation, senescence, apoptosis and functions of HBMEC were analyzed. For mechanism study, the epidermal growth factor receptor (EGFR)/ extracellular regulated protein kinases (ERK)/ p38 mitogen-activated protein kinase (p38 MAPK), and phosphatidylinositol-3-kinase (PI3K), serine/threonine kinase (Akt), endothelial nitric oxide synthase (eNOS), and p-Akt were analyzed. Results showed that: 1) MiR-125a-5p expression was reduced in ox-LDL treated HBMEC. 2) Overexpression of miR-125a-5p prevented HBMEC from ox-LDL induced apoptosis, senescence, ROS production, and NO reduction. 3) Overexpression of miR-125a-5p increased HBMEC proliferation, migration and tube formation, while decreased HBMEC adhesion to leukocytes, as well as counteracted the effects of ox-LDL on those functions. 4) The levels of EGFR/ERK/p38 MAPK pathway, PI3K/Akt/eNOS pathway, cleaved caspase 3 and adherent molecular ICAM-1 and VCAM-1 were associated with the effects of ox-LDL on these HBMEC functions. In conclusion, miR-125a-5p could counteract the effects of ox-LDL on various HBMEC functions via regulating the EGFR/ERK/p38 MAPK and PI3K/Akt/eNOS pathways and cleaved caspase 3, ICAM-1 and VCAM-1 expression.