Role of vasodilator stimulated phosphoprotein in resistin-induced proliferation and migration of human coronary endothelial cells / 医学研究生学报

ABSTRACT

Objective Resistin,also known as an adipose tissue-specific secretory factor,is involved in the development and progression of artherosclerosis by inducing dysfunction of endothelial cells,but its action mechanism has been rarely studied.The purpose of this study was to investigate the effect of resistin on the function of human coronary artery endothelial cells (HCAECs) and the underlying mechanisms.Methods We constructed the lentiviral vasodilator-stimulated phosphoprotein (VASP) shRNA interfering sequence (the LV-siVASP group) and a negative control vector (the LV-sicntr group),transfected HCAECs with VASP siRNA or control siRNA,and determined the VASP mRNA and protein expressions by quantitative RT-PCR and Western blot,respectively.We treated the HCAECs with resistin at the concentrations of 0,10,50,100,and 200 ng/mL followed by-detection of its effects on the proliferation and migration of the cells by MTT and Transwell chamber assay,respectively.We made comparisons between the LV-siVASP and LV-sicntr groups in the proliferation and migration of the HCAECs treated with 100 ng/mL resistin,the expression of vascular endothelial growth factor receptor-2 (VEGFR2) by immunofluorescent staining,and the activity of RhoA by pull-down assays.Results Compared with the LV-sicntr group,the LV-siVASP group showed significantly inhibited expressions of RNA (100％ vs [68.1±0.8]％,P＜0.05) and the VASP protein (100％ vs [59.3± 1.7] ％,P＜0.05) Treatment with resistin at 50-200 ng/mL markedly increased the proliferation of the HCAECs at 48 hours (P＜0.05) and induced a dose-dependent promotion of their migration at 24 hours (P＜0.05).No significant difference was observed in the expression VEGFR2 between the two groups,while the activity if RhoA was remarkably reduced in the LV-siVASP group ([41.3±3.1] ％) as compared with the LV-sicntr group (P＜0.05).Conclusion Resistin promotes the proliferation and migration of HCAECs,and the influence of VASP ablation on the action of resistin is associated with the decreased activity of RhoA.