Peptide OM-LV20 promotes arteriogenesis induced by femoral artery ligature via the miR-29b-3p/VEGFA axis

Background and aims Therapeutic arteriogenesis is a promising direction for the treatment of ischemic disease caused by atherosclerosis. However, pharmacological or biological approaches to stimulate functional collateral vessels are not yet available. Identifying new drug targets to promote and explore the underlying mechanisms for therapeutic arteriogenesis are necessary. Methods Peptide OM-LV20 (20 ng/kg) was administered for 7 consecutive days on rat hindlimb ischemia model, collateral vessel growth was assessed by H&E staining, liquid latex perfusion, and specific immunofluorescence. In vitro, we detected the effect of OM-LV20 on human umbilical vein endothelial cell (HUVEC) proliferation and migration. After transfection, we performed quantitative real-time polymerase chain reaction, situ-hybridization and dual luciferase reporters to assessed effective miRNAs and target genes. The proteins related to downstream signaling pathways were detected by Western blot. Results OM-LV20 significantly increased visible collateral vessels and endothelial nitric oxide synthase (eNOS), together with enhanced inflammation cytokine and monocytes/macrophage infiltration in collateral vessel. In vitro, we defined a novel microRNA (miR-29b-3p), and inhibition of miR-29b-3p enhanced the proliferation and migration of HUVEC as well as the expression of vascular endothelial growth factor A (VEGFA). OM-LV20 also promoted the migration and proliferation of HUVEC, and VEGFA expression was mediated via inhibition of miR-29b-3p. Furthermore, OM-LV20 influence the protein levels of VEGFR2 and Phosphatidylinositol3-kinase (PI3K)/AKT and eNOS in vitro and vivo. Conclusions Our data indicated that OM-LV20 enhanced arteriogenesis via the miR-29b-3p/VEGFA/VEGFR2-PI3K/AKT/eNOS axis, and highlight the application potential of exogenous peptide molecular probes through miRNA could constitute a effective therapeutic arteriogenesis in ischemic condition.