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Cyclooxygenase-2 Induction by High-density Lipoprotein in Human Umbilical Vein Endothelial Cells Depends on Sphingosine Kinase 2 and Sphingosine 1-phosphate Receptors

Author: XiongShengLin
Tutor: YiGuangHui
School: Nanhua University
Course: Pathology and Pathophysiology
Keywords: High-density lipoprotein Sphingosine kinase 2 1 - sphingosine phosphate Cyclooxygenase- 2 Co-immunoprecipitation
CLC: R543.5
Type: Master's thesis
Year: 2011
Downloads: 64
Quote: 0
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Abstract


Objective : 1 - sphingosine phosphate (sphingosine 1-phosphate, S1P) is phosphorylated sphingosine kinase (sphingosine kinase 1/2, SphK1 / 2) catalyzed synthesis of sphingosine . Primarily in plasma bound to HDL (high density lipoprotein, HDL) or albumin and activated platelets and red blood cells . And SphK2 is nuclear synthesis of 1 - sphingosine phosphate key enzyme synthesis 1 - sphingosine phosphate inhibits histone deacetylase (histone deacetylase, HDAC) activity, and promote gene transcription. Endothelial cells induces cyclooxygenase- 2 (cycloxygenase, COX-2) synthesis of prostaglandin I2 (prostaglandinc I 2 , PGI 2 ), the synthesis of PGI 2 < / sub> inhibition of platelet aggregation , dilation of blood vessels . This study investigated in human umbilical vein endothelial cells through SphK2 HDL , and 1 - sphingosine phosphate receptor pathway regulates the expression of COX-2 signaling mechanism . METHODS: Human umbilical vein endothelial cells , HDL different time and concentration process , qPCR and WB detect COX-2, CREB, Sphk2 expression . Human umbilical vein endothelial cells after transfection SiRNASphk2 , qPCR and WB detection HDL on COX-2 expression . Human umbilical vein endothelial cells were used JTE ( inhibition of S1P2), (PTX inhibition of Gi / o), SB203580 ( inhibiting P38MAPK), Staurosporine ( inhibition of PKC), U0126 ( inhibition of ERK1 / 2) inhibitor treatment , qPCR and WB detection HDL on COX-2 expression . Co-immunoprecipitation and Sphk2 detection P-CREB binding conditions for positioning with laser confocal observation . Results : HDL dose and time increased COX-2, SphK2 expression and the CREB phosphorylation after transfection SphK2siRNA levels of COX-2 expression decreased ; using JTE ( inhibition of S1P2), SB203580 ( inhibiting P38MAPK), Staurosporine ( inhibition of PKC), U0126 ( inhibition of ERK1 / 2) inhibitor treatment , HDL on the upregulation of COX-2 significantly inhibited ; co-immunoprecipitation and confocal and SphK2 display P-CREB binding . Conclusion : HDL through S1P2-G i / o - PKC-P38MAPK/ERK-SphK2-P-CREB way affect the expression of COX-2 . HDL activates endothelial cell nucleus sphingosine kinase and the CREB phosphorylation, which increases the expression of COX-2 .

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CLC: > Medicine, health > Internal Medicine > Heart, blood vessels ( circulatory ) disease > Vascular disease > Artery disease
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