Effects of KLF2 on ox-LDL induced expression of microRNA-146a and proinflammatory cytokines in human umbilical vein endothelial cells / 中华微生物学和免疫学杂志

ABSTRACT

Objective To investigate the effects of Kr üppel-like factor 2 ( KLF2 ) on oxidized low density lipoprotein (ox-LDL) induced expression of microRNA-146a (miR-146a) and proinflammatory cyto-kines (MCP-1 and IL-6) in human umbilical vein endothelial cells (HUVECs).Methods Human umbili-cal vein endothelial cells (HUVECs) were cultured and then stimulated with 50μg/ml ox-LDL for 24 hours. HUVECs were infected with adenovirus vectors over-expressing human KLF2 at an appropriate multiplicity of infection.KLF2-siRNA duplexes were transfected into HUVECs to silence the gene expression .HUVECs were collected at time points of 0 h, 3 h, 6 h, 12 h, 24 h and 48 h after infection.Real-time quantitative-PCR was performed to measure the expression of miR-146a at mRNA level.Silenced endogenous miR-146a using LNA-anti-miR-146 a was transfected into HUVECs with lipofectamine 2000 .The levels of MCP-1 and IL-6 in supernatants were detected by ELISA .Results KLF2 remarkably inhibited the expression of miR-146 a in unstimulated and ox-LDL-stimulated HUVECs in a time-dependent manner .The ox-LDL induced ex-pression of miR-146a, MCP-1 and IL-6 in HUVECs were significantly decreased by KLF2.Silenced expres-sion of miR-146a downr-egulated the ox -LDL induced expression of MCP-1 and IL-6 in HUVECs.Moreover, silenced miR-146 a could partly reverse the inhibitory effects of KLF 2 on ox-LDL induced expression of MCP-1 and IL-6 in HUVECs.Conclusion KLF2 inhibited ox-LDL induced expression of MCP-1 and IL-6 in HUVECs partly through down-regulating the expression of miR-146a.