The Effect Of Atorvastatin Atorvastatin On The Inflammation Induced By Ox-LDL

Cardiovascular and cerebrovascular diseases is one of the main reasons of death.Atherosclerosis is the pathological basis of cardiovascular and cerebrovascular diseases. With its morbidity and mortality increase, atherosclerosis is a current research focuses. For the pathogenesis of the disease, people proposed the fatty infiltration theory firstly, then proposed the formation of platelet aggregation and thrombosis theory, and lately people proposed the damage reaction theory.With the deepening of the research, put forward the theory that atherosclerosis is an inflammatory disease.ROSS revealed that atherosclerosis is an inflammatory disease of the arterial wall, which is the formed by the interaction of vascular endothelial cells, mononuclear cells, and vascular smooth muscle cells. Atherosclerosis Characterized by the appearance of monocytes, macrophages and T lymphocytes in atherosclerotic plaque. Inflammatory response plays an important role throughout the whole process of atherosclerosis in cluding occurrence, development and cause of cardiovascular events.In the initial stages of atherosclerosis, many factors can cause inflammation of the arterial wall, where ox-LDL is an important proinflammatory substances.It can cause the expression of many inflammatory genes. The products of these gene such as adhesion molecules and chemokines can make local monocyte accumulation and activation. Monocyte can migrate into the subendothelial and secrete some cytokines. Then it further promote the development of atherosclerosis.Statins is the inhibitor of3-hydroxy3-methyl glutaryl coenzyme A reductase.It can lower blood cholesterol levels and it is the most effective drug in the treatment of hyperlipidemia and atherosclerosis. More and more evidence showed that statins can reduce cardiovascular disease morbidity and mortality not only attributing to the lipid-lowering effect, but also attributing to many other pharmacological effects. These effects includes improving the function of vascular endothelial, promoting fibrinolysis, anti thrombosis etc. In recent years, the research on the anti-inflammatory effect of statins is increasing. The discovery anti-inflammatory effect of statins showed that statins may have other effects besids the lipid-lowering effect to reat the atherosclerosis. But the anti-inflammatory mechanism of statins is still unclear. In this study, we investigated wether ox-LDL can induce inflammatory reaction in human umbilical vein endothelial cells (HUVEC) and mononuclear cells (THP1), and further analyzed wether atorvastatin can reduce the inflammatory reaction induced by ox-LDL.Part1The effect of atorvastatin on the infammation induced by ox-LDL in HUVEC cellsObjective:To examine whether ox-LDL can induce the expression of ICAM-1, VCAM-1and MCP-1in HUVEC cells and analyzed the effect of atorvastatin on the.the expression of ICAM-1,VCAM-1and MCP-1induced by the ox-LDL.Methods:1. The culture of HUVEC cells:Recovery of HUVEC cell, cultured in vitro using ECM medium, observed the morphology and proliferation of the cells every day.2. The effect of ox-LDL on the proliferation of HUVEC cells was detected by MTT3. Experiment groups:(1)the control group:add medium, as the control;(2)0x-LDL group (100μg/mL), only adding ox-LDL;(3) Atorvastatin group (1μM), just add the atorvastatin(4) Atorvastatin (1μM)+ox-LDL(100μg/mL) group.4. The mRNA expression of TNF-a, IL-6and IL-1β was detected by Real-Time PCR5. The protean expression changes of ICAM-1and VCAM1in HUVECs cells was detected by Western-Blot. 6. The changes of MCP-1secretion was determined by ELISA analysis.Results:1. The results of MTT showed that the growth and proliferation of HUVEC cells was not affected by0.5,1,2.5,5μ M ox-LDL.2.Western-Blot assay showed that, compared with the control group, ox-LDL can enhance the protein expression of ICAM1and VCAM1, and atorvastatin can decrease protein expression of ICAM1and VCAM1induced by ox-LDL3. The results of Real-Time PCR revealed that ox-LDL can increase the mRNA expression of ICAM1, VCAM1and MCP-1, and atorvastatin can decrease the mRNA expression of ICAM1, VCAM1and MCP-1induced by ox-LDL.4. ELISA assay showed that ox-LDL can increase the secretion of MCP-1, and atorvastatin can decrease the MCP-1secretion induced ox-LDL.Conclusion:The data indicate that ox-LDL can induce inflammation in HUVEC cells, and atorvastatin can reduce the inflammation induced by ox-LDL.Part2The effect of Atorvastatin on the infammation induced by ox-LDL in THP1cellsObjective:To examine wether ox-LDL can induce the expression of TNF-α、IL-6and IL-1βin THP1cells and analyzed the effect of atorvastatin on the.the expression of TNF-α、IL-6and IL-1β induced by the ox-LDL.Methods:1.The culture of THP1cells:Recovery of THP1cell, cultured in vitro using RPMI-1640medium, and observed the morphology and proliferation of the cells every day.2. The effect of ox-LDL on the proliferation of THP1cells was detected by MTT3. Experiment groups:(1)the control group:add medium, as the control;(2)Ox-LDL group (100μg/mL), only adding ox-LDL;(3) Atorvastatin group (1μM), just add the atorvastatin(4) Atorvastatin (1μM)+ox-LDL(100μg/mL) group.4. The mRNA expression of TNF-a, IL-6and IL-1β was detected by Real-Time PCR5. The changes of TNF-a, IL-6and IL-1β secretion was determined by ELIS A analysis.Results:1. The results of MTT showed that the growth and proliferation of THP1cells was not affected by0.5,1,2.5,5μM ox-LDL.2.The results of Real-Time PCR revealed that ox-LDL can increase the mRNA expression of TNF-a, IL-6and IL-1β, and atorvastatin can decrease the mRNA expression of TNF-a, IL-6and IL-1β induced by ox-LDL.3. ELIS A assay showed that ox-LDL can increase the secretion of MCTNF-a, IL-6and IL-1β, and atorvastatin can decrease theTNF-a, IL-6and IL-1p secretion Induced by ox-LDL.Conclusion:The data indicate that ox-LDL can induce inflammation in THP1cells, and atorvastatin can reduce the inflammation induced by ox-LDL.