Simvastatin Modulates Inflammatory Mediat-Ors Expression In Human Umbilical Vein Endothelial Cells

Objective:Statins are potent inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase.These drugs are capable of lowering the serum levels of cholesterol and are successfully used to treat hypercholesterolemia and atherosclerosis.Moreover,the ability of statins to reduce the mortality and morbidity of cardiovascular diseases has been ascribed not only to their cholesterol-lowering activities but also to a number of additional effects,including improved endothelial cell function,enhanced fibrinolysis, and antithrombotic activity.In addition,a number of important anti-inflammatory effects of statins have been reported.The anti-inflammatory effects of statins implicated more clinical benefit that can be obtained in the treatment of atherosclerosis.Little is known about the mechanisms by which statins counteract inflammation.Interleukine 6(IL-6),interleukine 8(IL-8),tumor necrosis factor (TNFα),urokinase plasminogen activator/urokinase plasminogen activator receptor(uPA/uPAR)are essential inflammatory mediators which play important roles in the inflammation process.They can be expressed from endothelial cells which induced by injurious factors to active the leukocyte and infhence the process of chemotactic,adhesion and transmigration.We use the inflammatory stimulating factor-Lipopolysaccharides (LPS)on the human umbilical vein endothelial cell(HUVEC)to induce the mRNA exprssion of IL-6,IL-8,TNFαand uPA/uPAR.Then we observe the influence of simvastatin on the mRAN expression of HUVEC.The results can be used to improve the theory of inflammatory mechanism of statins.Method:1.Cell culture:Collect the fresh umbilical core,cultivate the umbilical vein endothelial cells,identified,passage,the cells of 2-3 era could be use.2.Divide into four groups random:①cntrol group;②LPS group(100ng/ml);③simvastatin group(5μg/ml);④LPS and simvastatin group(10μg/ml).The four groups of cells were co-incubated for 1-24 hours(1h,12h,24h).3.The levels of IL-6,IL-8,TNFαin the collected culture supernatants were measured by the ELISA kit.4.The mRAN expression of IL-6,IL-8,TNFαand uPA/uPAR of the HUVEC were detected by the real-time quantitative PCR(RT-PCR).5.The experiment above were repeated three times,then use software to do the statistical anlysis.Result:1.The identification of HUVEC:The morphology identification,in the microscope,the endothelial cells distribute uniformity,grow in monolayer,polygon,arrays like cobblestone-appearance.FactorⅧantigon immunofluorescence detection,in the fluorescent microscope, there's flavovirens fluor in the endochylema.2.Theresult of ELISA:The production and secretion of IL-6,IL-8,TNFαin HUVEC s were significantly induced by LPS treatment(p＜0.01);the elevated level of these cytokines was reduced significantly by simvastatin treated cells when compared with control group(p＜0.01)。3.The result of real time quantitative PCR:The mRNA expression of IL-6,IL-8,TNFα,uPA/uPAR in HUVECs were significantly induced by LPS treatment(p＜0.05);The mRNA expression of IL-6,IL-8,TNFα,uPA/uPAR in HUVECs reduced significantly by simvastatin treated cells when compared with control group(p＜0.05).Conclution:1.The mRNA expression of IL-6,IL-8,TNF-α,uPA/uPAR in HUVECs can be induces by LPS.2.Simvastatin can reduce the mRNA expression of IL-6,IL-8,TNF-α,uPA/uPAR in the HUVECs stimulaed by LPS.