| Background Cardiovascular disease caused by atherosclerosis is a common disease damaging people's health.Ross thought atherosclerosis is a chronic inflammatory caused by injury factors including oxidative low density lipoprotein(Ox-LDL).we hypothesized that Ox-LDL may induce mRNA and protein expression of lectin-like oxidized LDL recepter-1(LOX-1).The non-lipid effects of statins such as anti-infammotory,stabilizing plaque ect are increasingly recognized.The present study was undertaken to demonstrate the effect of atorvastatin on LOX-1 protein and LOX-1 mRNAAim To observe the influence of atorvastatin(ATV),a kind of 3-Hydroxy-3-methylglutary coenzyme A(HMG COA)reductase inhibitors(statins),at different concentrations on the mRNA and protein expression of LOX-1 in cultured human umbilical vein endothelial cells(HUVEC) activated by Ox-LDL in order to understand the non-lowing lipid of atorvastatin. First Part The expression of LOX-1 mRNA and Protein in HUVECs activated by Ox-LDLMethodsLow density lipoprotein(LDL)is isolated from the blood by one-step discontinuous density gradient ultracentrifugation.Oxidation was carried out with Cu2+.Human umbilical vein endothelial cells(HUVEC)were isolated from freshly obtained human umbilical cords by collagenase typeⅡtreatment.The cells were cultured in M1640 supplemented with 20%bovine colf serum and passages 2 to 3 were used in this study.HUVECs were coincubated with different concentrations(20mg/L,40mg/L,60mg/L,80mg/L)of Ox-LDL for 24h or with 40 mg/L Ox-LDL for different periods(3,6,12,24,48h).LOX-1 mRNA was detected by quantitative reverse transcription polymerase chain reaction(RT-PCR).The protein of LOX-1 was detected by cell-based enzyme linked immunosorbant assay.ResultsThe levels of mRNA and protein of LOX-1 increased initially after cells were coincubated with 20mg/L Ox-LDL(P＜0.01),and reached peak value at 80mg/L,and then declined;In the same concentration of 40 mg/L Ox-LDL.the expression of mRNA and protein of LOX-1 were increased significantly with the coincubation time from 3h to 24h(P＜0.01),and maintained high level to 48h.Second Part The inhibition effect of atorvastatin on the mRNA and Protein Expression of LOX-1 in HUVECs activated by Ox-LDLMethodsThere were three groups:control,Ox-LDL and atorvastatin group.Cells of Ox-LDL groups were coincubated with 40mg/L Ox-LDL for 24h.Cells of atorvastatin group were firstly coincubated with 0.05,0.1,1.0,10μmol/L atorvastatin respectively for 1h,then coincubated with 40mg/L Ox-LDL for 24h.LOX-1 mRNA was detected by RT-PCR..The protein of LOX-1 was detected by cell-based enzyme linked immunosorbant assay.ResultsThe expression of LOX-1 mRNA and protein of Ox-LDL group and atorvastatin group was more than that of control(P＜0.01).The expression of LOX-1 mRNA and protein of atorvastatin groups were less than that of Ox-LDL group(P＜0.05)Conclusions1.Ox-LDL upregulates the expression of the mRNA and protein of LOX-1 in a concentration-dependent and time-dependent manner.2.Atorvastatin inhibits the expression of the mRNA and protein of LOX- 1 in HUVECs activated by Ox-LDL in a concentration-dependent manner.3.Atorvastatin may increase the stability of plaques and decelerate the progression of atherosclerosis... |
