The mechanism of blood‐brain barrier (BBB) disruption, involved in poststroke edema and hemorrhagic transformation, is important but elusive. We investigated microRNA‐150 (miR‐150)‐mediated mechanism in the disruption of BBB after stroke in rats. We found that up‐regulation of miR‐150 increased permeability of BBB as detected by MRI after permanent middle cerebral artery occlusion in vivo as well as increased permeability of brain microvascular endothelial cells after oxygen‐glucose deprivation in vitro. The expression of claudin‐5, a key tight junction protein, was decreased in the ischemic boundary zone after up‐regulation of miR‐150. We found in brain microvascular endothelial cells that overexpression of miR‐150 decreased not only cell survival rate but also the expression levels of claudin‐5 after oxygen‐glucose deprivation. With dual‐luciferase assay, we confirmed that miR‐150 could directly regulate the angiopoietin receptor Tie‐2. Moreover, silencing Tie‐2 with lentivirus‐delivered small interfering RNA reversed the effect of miR‐150 on endothelial permeability, cell survival, and claudin‐5 expression. Furthermore, poststroke treatment with antagomir‐150, a specific miR‐150 antagonist, contributed to BBB protection, infarct volume reduction, and amelioration of neurologic deficits. Collectively, our findings suggested that miR‐150 could regulate claudin‐5 expression and endothelial cell survival by targeting Tie‐2, thus affecting the permeability of BBB after permanent middle cerebral artery occlusion in rats, and that miR‐150 might be a potential alternative target for the treatment of stroke.—Fang, Z., He, Q.‐W., Li, Q., Chen, X.‐L., Baral, S., Jin, H.‐J., Zhu, Y.‐Y., Li, M., Xia, Y.‐P., Mao, L., Hu, B. MicroRNA‐150 regulates blood‐brain barrier permeability via Tie‐2 after permanent middle cerebral artery occlusion in rats. FASEB J. 30, 2097–2107 (2016). www.fasebj.org