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Modification de l'expression et de la phosphorylation de la VASP dans les cellules endothéliales soumises aux écoulements

Abstract : Vasodilator stimulated phosphoprotein (VASP) is a member of Ena/VASP protein family that links to the regulation of actin filaments and to signal transduction during cell morphological modification and migration. VASP tightly associates with microfilament, in particular at focal adhesions and celllamellipodia. ln this study, we examined the expression, phosphorylation and redistribution of VASP in HUVECs under flow conditions. And the mechanism of VASP phosphorylation was further studied with the enzyme inhibitors. We found by double immunofluoreseent staining that after exposure to a shear of 1,0 Pa for 24h, spots of VASP were concentrated at the two extremities ofthick stress fibres. RT-PCR results indicated that the shear could induce VASP mRNA increase. Western blotting results showed that there was only non-phosphorylated VASP in static cells. The VASP phosphorylation appeared as a transient phenomenon with peak values after Ih and 4h exposures to the same shear stress. The shear stress of 1,5 Pa had similar tendencyas that of 1,0 Pa while the shear stress of 0,2 Pa showed a lower effect. Additionally, inhibition of the actin polymerization by cytoehalasin D blocked VASP expression and phosphorylation. H89, à PKA inhibitor, inhibited shear-induced increase of VASP expression and phosphorylation, while L-NNA, an NOS inhibitor showed adverse effects.These results show that shear stress induces the phosphorylation of VASP in endothelial cells, and this phosphorylation depends on the actin integrity and via a cAMP/PKA pathway.
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Lei Wei. Modification de l'expression et de la phosphorylation de la VASP dans les cellules endothéliales soumises aux écoulements. Biotechnologies. Université Henri Poincaré - Nancy 1, 2004. Français. ⟨NNT : 2000NAN10254⟩. ⟨tel-01746959⟩



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