The Plasma Exosome MiR-144-3p In Crohn’s Disease Affects Vascular Endothelial Cell Function By Inhibiting FN1

Background: Crohn’s disease(CD)is a chronic nonspecific inflammatory disease with unknown pathogenesis and vascular changes associated with the progression of CD.Many studies have shown that miRNAs participate in the development of CD.However,the effect of miRNAs in plasma exosomes on vascular endothelial cells in CD has not been investigated.Our study aimed to identify the differential miRNAs in plasma exosomes in CD and explore their potential roles in human umbilical vein endothelial cells(HUVECs).Methods:1.In our study,exosomes were extracted from circulating blood to identify differential miRNAs.2.After in vitro transfection of HUVECs with miR-144-3p mimics and inhibitors and the corresponding controls,cell counting kit-8,wound-healing,Transwell migration,and tube formation assays were performed to study the viability,migration,and angiogenesis of HUVECs.3.Furthermore,four bioinformatics analysis was used to predict miRNA targets,q RT-PCR and Western blot analysis were used to determine FN1 protein expression.4.In addition,exogenous supplementation with the fibronectin 1(FN1)protein rescued the effects of miR-144-3p on change in cell function in vitro.Results:1.miR-144-3p was significantly increased in the exosomes of patients with CD compared with those in the control group.2.The promotion or inhibition of miR-144-3p correspondingly abolished or accelerated cell viability,migration,and angiogenesis.3.We predicted all target genes of miR-144-3p by website,and then found that the target genes of miR-144-3p were mainly enriched in macromolecule-related signaling pathways by GO with KEGG analysis.miR-144-3p could inhibit the expression of FN1 by q RT-PCR and Western blot experiments.4.Exogenous FN1 can increase the viability,migration,and angiogenesis of HUVECs in vitro.An exogenous increase in FN1 protein ameliorated the reduced viability,migration,and angiogenesis of HUVECs caused by transfection with miR-144-3p.Conclusions: miR-144-3p was significantly increased in CD plasma exosomes,and miR-144-3p could lead to vascular endothelial cell dysfunction by inhibiting the expression of FN1.