A Study On The Role And Mechanism Of TSLP In Atherosclerosis

Part Ⅰ. Expression of thymic stromal lymphopoietin in atherosclerotic vessels.Objective To observe whether TSLP was expressed in atherosclerotic vessels of human and atherosclerosis model mice, and exposit its relationship with dendritic cells.Methods Normal groups were no-atherosclerotic vessels from human and18-week-old C57BL/6male mice. Positive groups were atherosclerotic vessels taken from human and18to22week old ApoE-deficient mice on the C57BL/6background (ApoE-KO mice). Immunohistochemistry was used to detect the expressions of TSLP, CDllc and CD4in blood vessels. In situ hybridization technical inspection was pursued to show TSLP mRNA in vessels. Double immunofluorescence staining was used to display the relationship between TSLP and dendritic cells.Results Normal groups expressed few TSLP. Positive groups of human and ApoE-KO mice released significant TSLP protein and TSLP mRNA. In atherosclerotic vascular walls, dendritic cells located around the cells secreted TSLP. Immunohistochemical single stain for TSLP, CD1lc and CD4in serial sections showed that DCs and TCs enfolded the cells secreted TSLP.Conclusion TSLP were obviously detected in atherosclerotic vessels of human and ApoE-KO mice, and were gathered around dendritic cells. It probably showed that, as the upstream impact factor, TSLP directly involved in the immune inflammation progress of atherosclerosis. Part Ⅱ. Expression of thymic stromal lymphopoietin in cultured vascular endothelial cells induced by ox-LDL.Objective To survey the expression of thymic stromal lymphopoietin (TSLP) in primary human umbilical veins endothelial cells (HUVECs) induced by ox-LDL.Methods Cultured HUVECs were divided into the control group and ox-LDL (12.5,25,50,100mg/L) treated groups. TSLP protein and mRNA were inspected by ELISA, immunofluorescence and real-time RT-PCR from primary HUVECs cultures.Results Few TSLP was expressed in primary HUVECs. Significant increases of TSLP mRNA and protein release were detected in HUVECs incubated with ox-LDL.Conclusion The expression of TSLP in vascular endothelial cells can be induced by ox-LDL, which may play a part in vascular immunoinflammation.Part Ⅲ. Expression of thymic stromal lymphopoietin in cultured vascular smooth muscle cells induced by ox-LDL.Objective To survey the expression of thymic stromal lymphopoietin (TSLP) in human vascular smooth muscle cells (HVSMCs) induced by ox-LDL.Methods Cultured HVSMCs were divided into ox-LDL(12.5,25,50,100mg/L) treated groups and the control group. TSLP protein and mRNA were inspected by ELISA, immunofluorescence and real-time RT-PCR from HVSMCs cultures.Results Few TSLP was expressed in normal HVSMCs. Significant increases of TSLP mRNA and protein release were detected in HVSMCs incubated with ox-LDL.Conclusion The expression of TSLP in vascular smooth muscle cells can be induced by ox-LDL, which may play a critical role in vascular immunoinflammation. Part Ⅳ. Atorvastatin inhibited the expression of thymic stromal lymphopoietin in cultured vascular endothelial cell induced by ox-LDL.Objective To explore effect of atorvastatin on expression of thymic stromal lympho-poietin (TSLP) in primary HUVEC induced by ox-LDL.Methods Cultured HUVECs with ox-LDL (50mg/L) were divided into three groups:the control group, atorvastatin (1μmol/L) treated group and atorvastatin (10μmol/L) treated group. TSLP protein and mRNA were assessed by ELISA, real-time RT-PCR, and immunofluorescence from primary HUVEC cultures at six hours and12hours.Results The control group expressed high-level TSLP. Incubation of HUVECs with atorvastatin resulted in a significant decrease of TSLP mRNA and protein release from HUVEC, which is in a dose-dependent manner.Conclusion Atorvastatin can inhibited the expression of TSLP in cultured vascular endothelial cells induced by ox-LDL, which may be one of the mechanisms that statins inhibit inflammation in atherosclerosis.