Activin A And TNF-? Regulate The Migration And Tube Formation Of Human Umbilical Vein Endothelial Cells In Vitro

Angiogenesis is involved in the development and progression of most diseases,including tumors,inflammation,and atherosclerosis.Activin A,a member of the transforming growth factor-?(TGF-?)superfamily,is considered to be a factor regulating endothelial cell growth and differentiation in vitro and angiogenesis in vivo.Tumor necrosis factor-?(TNF-?)is an important inflammatory factor produced by a variety of cells and plays an important role in cell proliferation,migration,invasion and angiogenesis.The previous study of the research group found that activin A and TNF-? mainly affect the activity of fibroblasts through mutual antagonism.Therefore,human umbilical vein endothelial cells(HUVEC)were selected in this study to investigate the effects of activin A and TNF-? on HUVEC cell activity,migration,invasion and tube formation and other biological behaviors,providing potential therapeutic targets for further exploration of inflammation,diabetes,cardiovascular diseases and other angiogenesis related diseases.?.METHODS1.Cell viability was detected by MTT assay.2.The level of IL-6 in cell culture supernatant was determined by ELISA.3.Cell scratch assay and Transwell chamber assay were used to detect cell migration and invasion in vitro,and matrix tube formation assay was used to detect cell tube formation.4.RT-PCR and Western blotting were used to detect the expression of genes and proteins related to cell signaling and cell migration.?.RESULTS1.Both activin A and TNF-? inhibited HUVEC cell viability.2.Activin A induced IL-6 secretion in HUVEC cells,but decreased IL-6 production in HUVEC cells induced by TNF-?.PCR detection of IL-6 m RNA in HUVEC cells was consistent with ELISA detection.3.Activin A inhibited HUVEC cell migration,invasion and tube formation,and TNF-?-induced HUVEC cell migration,invasion and tube formation.4.Activin A had no significant effect on the expression of MMP-2,MMP-9 and N-cadherin protein in HUVEC cells,but activin A inhibited the expression of MMP-2,MMP-9 and N-cadherin protein in HUVEC cells induced by TNF-?.5.LMT-28,an IL-6 inhibitor,could antagonize the inhibitory effect of Activin A on HUVEC cell migration,but had no effect on TNF-? induced HUVEC cell migration.6.Both activin A and TNF-? could increase the expression of PSmad3 and P-STAT3 proteins in HUVEC cells.When treated with activin A and TNF-?,activin A partially inhibited the TNF-?-induced elevated expression of P-Smad3 and P-STAT3 proteins in HUVEC cells.?.CONCLUSION1.Activin A regulates HUVEC cell activity through the classical Smad pathway.2.Activin A inhibits TNF-?-induced HUVEC cell migration,which may be related to IL-6 /STAT3 signaling pathway.3.Activin A inhibits TNF-?-induced angiogenesis in HUVEC cells,suggesting that activin A may be involved in the regulation of angiogenesis and is expected to be a therapeutic target for tube formation-related diseases.