Dual Targeting of Angipoietin-1 and von Willebrand Factor by microRNA-671-5p Attenuates Liver Angiogenesis and Fibrosis

Angipoietin-1 (Angpt1) and von Willebrand factor (VWF) are two important angiogenic molecules that can drive pathologic angiogenesis and progression of liver fibrosis in our previous study. MicroRNAs (miRs) participate in a variety of physiological and pathological processes, including angiogenesis. However, the critical miRs targeting Angpt1 or VWF and potential molecular mechanism underlying liver fibrosis-associated angiogenesis is not clear yet. Human liver tissues were obtained from patients with different chronic liver diseases. Mouse models of liver fibrosis were induced by injection of CCl₄ or bile duct ligation (BDL) operation. MiR-671-5p was predicted to target Angpt1 and VWF from three databases (miRanda, RNA22v2, and miRwalk). MiR-671-5p expression was decreased in the fibrotic liver of human and mice, with a negative correlation with the levels of Angpt1, VWF, sphingosine kinase-1 (SphK1, the rate-limiting enzyme for sphingosine 1-phosphate [S1P] formation), transforming growth factor β1 (TGFβ1), hypoxia inducible factor (Hif)1α, Hif2α, and fibrosis markers. Importantly, miR-671-5p expression was down-regulated in fluorescence-activated cell sorted liver sinusoidal endothelial cells and hepatic stellate cells (HSCs) in CCl₄ mice compared with control mice. In vitro miR-671-5p expression was also decreased in S1P-stimulated HSCs and TGFβ1-activated liver sinusoidal endothelial cells, negatively correlated with Angpt1 and VWF expression. MiR-671-5p directly targeted Angpt1 and VWF by luciferase reporter assays. In vivo administration of miR-671-5p agomir decreased the messenger RNA and protein levels of Anpgt1 and VWF, and attenuated CCl₄ -induced or BDL-induced liver angiogenesis and fibrosis. Conclusion: We identify the negative regulation of miR-671-5p on Angpt1 and VWF and liver fibrosis-associated angiogenesis, which may provide promising targets for the prevention and treatment of liver disease.