Regulation Of MicroRNA-33 On Ox-LDL-Induced Inflammatory Response In Human Umbilical Vein Endothelial Cells

Objective:1.To compare the changes of mi R-33 and NF-?BP65 induced by ox-LDL in HUVEC;2.To investigate the effect of mi R-33 inhibitor on ox-LDL-induced inflammatory response in HUVEC and to reveal the regulation of micro RNA-33 on inflammatory response of HUVEC,further elucidation of the role of mi R-33 in the pathogenesis of atherosclerosis.Methods: The first part of the experiment: HUVEC were treated with 50?g/m L,100 ?g/m L and 200 ?g/m L ox-LDL,respectively.After 24 hours,q PCR was used to detect the difference of mi R-33 expression level;Western Blot was used to check NF-?BP65 expression level.Part II of the experiment: Cultured HUVEC,grouped:(1)normal(2)100 ?g/m L ox-LDL(3)100 ?g/m L ox-LDL+NC(4)100 ?g/m L ox-LDL+mi R-33 inhibitor,transfected(3)groups with 5pmol NC and 5pmol mi R-33 inhibitor was transfected into(4)groups,(2)(3)(4)endothelial cells were treated with 100 ?g/m L of ox-LDL 6 hours later.After 24 hours,the following tests were performed: ELISA was used to detect inflammatory factors in culture medium,Western Blot was used to detect NF-?BP65expression,and q PCR was used to detect the expression level of mi R-33 in cells.Results: 1.50 ?g/m L(p<0.05),100 ?g/m L(p<0.01)and 200 ?g/m L(p<0.01)concentrations of ox-LDL stimulated the expression of mi R-33 in HUVEC compared with the blank control group,and with the increase of ox-LDL concentration,The expression of mi R-33 and NF-?BP65 was gradually increased.2.In the human umbilical vein endothelial cells stimulated by ox-LDL,the expression of pro-inflammatory cytokines TNF-? and IL-6 was significantly inhibited in the inhibitor group of mi R-33 compared with NC group(p<0.01),The expression of mi R-33 was significantly down-regulated(p<0.01)and the concentration of NF-?BP65 was decreased.Conclusion: 1.ox-LDL can up-regulate the expression of mi R-33 and NF-?BP65 in human umbilical vein endothelial cells,and the up-regulation of this expression is concentrating on ox-LDL concentration.2.The mi R-33 inhibitor may reduce inflammatory response by down-regulating the expression of NF-?BP65 in human umbilical vein endothelial cells.