RESUMO
Coronary artery bypass graft (CABG) is one of the primary methods of treating coronary heart disease (CHD); however, vein graft restenosis is a major limiting factor of the effectiveness of CABG. Emerging evidence has indicated that miR423 is associated with vascular diseases. Additionally, upregulation of a disintegrin and metalloproteinase with thrombospondin motifs7 (ADAMTS7) contributes to neointima formation by promoting the proliferation and migration of vascular smooth muscle cells and inhibiting the proliferation and migration of endothelial cells. The aim of the present study was to examine the effects of miR423 target, ADAMTS7, on regulating vein graft disease and identify novel biomarkers for use in therapy of vein graft failure (VGF). Aberrant expression of miR423 in plasma of patients with CHD prior to and following CABG confirms that miR423 may be a suitable target for preventing VGF. Furthermore, a dualluciferase reporter gene assay indicated that miR423 directly interacted with ADAMTS7 and suppressed its expression. Ectopic expression of miR423 suppressed ADAMTS7, resulting in decreased proliferation and migration rates of human umbilical vein smooth muscle cells by targeting ADAMTS7, but resulted in increased proliferation and migration of human umbilical vein endothelial cells in vitro. Overexpression of miR423 also enhanced reendothelialization and decreased neointimal formation in a rat vein graft model. In conclusion, the results of the present study demonstrated that the miR423/ADAMTS7 axis may possess potential clinical value for the prevention and treatment of restenosis in patients with CHD following CABG.
Assuntos
Proteína ADAMTS7/genética , Oclusão de Enxerto Vascular/genética , MicroRNAs/genética , Animais , Movimento Celular , Proliferação de Células , Ponte de Artéria Coronária , Doença das Coronárias/genética , Doença das Coronárias/cirurgia , Regulação para Baixo , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Pessoa de Meia-Idade , Ratos Sprague-Dawley , Regulação para CimaRESUMO
BACKGROUND: Fibronectin (FN) plays vital roles in cell adhesion, differentiation, proliferation and migration. It is involved in the process of embryonic development and is highly conserved during evolution. The EIIIA and EIIIB of FN show a very high degree of homology among vertebrates. Embryos deleting both EIIIA and EIIIB displayed multiple embryonic cardiovascular defects, implying their crucial role during embryogenesis. The correlation of spliced EIIIB, EIIIA, and IIICS of FN to heart development was studied by observing their chronological expression in mice heart. METHODS: C57 mice embryos at E11.5, E12.5, E13.5, E14.5, E15.5, E16.5, E17.5, E18.5, E19.5 days, postnatal day 1 (P1d), and adult male mice (3 months) were used. For each alternatively spliced FN1 domain (EIIIB, EIIIA and IIICS), primer pairs were designed for specific amplification. Total RNA was extracted from the heart tissue, reverse transcripted to cDNA, followed by RT-PCR with specific primers. The PCR amplification was verified by agarose gel electrophoresis, showing specific fragments of the expected sizes. RESULTS: In adult mice heart, only alternatively splice variants of EIIIA-, EIIIB-, IIICS+ were expressed. While in embryonic mice, spliced variant of EIIIA+/-, EIIIB+/-, IIICS+ were observed. The expression of EIIIA and EIIIB changed during heart development. CONCLUSIONS: FN is crucial for the normal development of the embryonic heart by modulating cardiac neural crest (CNC) proliferation and survival, and maintenance of CNC cells. FN1 gene seems to play a significant role by expression of highly conserved EIIIA and EIIIB in embryonic heart development.