MiR-587 Promotes The Growth Of Human Aortic Endothelial Cells By Regulating The Expression Of TLR4

Objective: By detecting the expression of mir-587 and TLR4 in human aortic endothelial cells induced/uninduced by ox-LDL.The expression of miR-587 was up-regulated or down-regulated in the cells,and how miR-587 affected the growth of human aortic endothelial cells by regulating the expression of TLR4 was explored.Methods: The expression of miR-587 and TLR4 in human aortic endothelial cells induced by ox-LDL was detected by real-time PCR.MTT assay was used to detect the effect of miR-587 on cell proliferation.The effect of miR-587 on apoptosis was analyzed by JC-1 experiment.After miRdb software was used to analyze and predict the possible binding sites of miR-587 and TLR4,luciferase reporter gene experiment was used to detect the targeted regulatory effect of miR-587 on TLR4,and to analyze whether the overexpression of miR-587 affects the activity of TLR4.To construct human aortic endothelial cell line with over or under expression of miR-587,and to detect the regulatory effect of over or under expression of miR-587 on TLR4 and caspase-8 protein by Western blot.TLR4 was transfected into human aortic endothelial cells overexpressing miR-587.Cell growth was detected by MTT,apoptosis was detected by JC-1,and expression of TLR4 and caspase-8 was detected by Western blot.Statistical methods were used to compare the single factor statistical differences among the groups.Results: Real time PCR indicated that ox LDL could down regulate the expression of miR-587 and promote the expression of TLR4.MTT assay showed that the proliferation of human aortic endothelial cells was significantly promoted after mi R-587 was transfected.JC-1 showed that miR-587 could inhibit the apoptosis of human aortic endothelial cells.Mirdb software predicted that miR-587 had binding sites with the3'UTR region of TLR4,and luciferase reporter gene showed that transfection of miR-587 could significantly down regulate the activity of TLR4.Western blot showed that over expression of miR-587 could inhibit the expression of TLR4 and caspase-8.Transfection of TLR4 could reduce the proliferation and apoptosis of the cells,and the down-regulation of TLR4 and caspase-8.Conclusions: miR-587 can promote the growth of human aortic endothelial cells byinhibiting TLR4.