The Influence Of Angiotensin-(1-7) On The Secretion Of E-Selectin And Monocyte Chemoattractant Protein-1 In Cultured Human Umbilical Vein Cells Induced By Angiotensin Ⅱ

Background & ObjectivesAngiotensin-(1-7)[Ang-(1-7)],a bioactive peptide in the renin-angiotensin system,has both vasodilatory and antiproliferative activities that are opposite those of angiotensinⅡ(AngⅡ).Recent studies suggest that atherosclerosis is a chronic inflammatory disease.E-selectin (E-sel) is a critical adhesion molecule existing on vascular endothelial cells,which promotes leucocyte adhering to endothelial cells to initiate atherosclerosis.Many evidences suggest that monocyte chemoattractant protein 1(MCP-1) appear to play an early and important role in the recruitment of monocytes to atherosclerotic lesions.However,it remains unknown whether Ang-(1-7) can prevent the increase of E-sel and MCP-1 in human umbilical vein Cells(HUVEC) induced by AngⅡ.And it is also unknown whether it is acted through its specific receptor Mas.To explore the influence of angiotensin-(1-7) on the secretion of E-selectin(E-sel) and monocyte chemoattractant protein-1(MCP-1) in cultured human umbilical vein Cells Induced by AngiotensinⅡ(AngⅡ).Methods1.Culture and Identification of HUVECs.2.Group:(1) Control group:the HUVECs were cultured in serum-free RPMI1640 without any stimulators or inhibitors;(2) AngⅡgroup:the HUVECs were stimulated with the final concentration AngⅡ100nmol/L;(3) Ang-(1-7) group:the HUVECs were treated with the final concentration Ang-(1-7) 1000nmol/L;(4) Ang-(1-7) + AngⅡgroup:the HUVECs were pretreated with the final concentration Ang-(1-7) 10nmol/L;Ang-(1-7) 100nmol/L; Ang-(1-7) 1000nmol/L;Ang-(1-7) 10000tool/L;and after 30min adding the final concentration AngⅡ100nmol/L;(5) A-779 + Ang-(1-7) + AngⅡgroup:the HUVECs were pretreated with the certain A-779 of 1000umol/L for 30 minute,then pretreated with the final concentration Ang-(1-7) 1000umol/L for 30 minute,finally stimulated with the final concentration AngⅡ100nmol/L.3.The HUVECs E-sel and MCP-1 protein expression were measured by enzyme linked immunosorbent assay(ELISA),and E-sel and MCP-1 mRNA expression were detected by reverse transcription polymerase chain reaction(RT-PCR).Results1.The HUVECs arrayed like pitching stone under light microscopy;Ⅷfactor in HUVECs showed positive reaction by immunohistochemistry.All these cells were identified as ECs.2.Compared with the medium group,100nmol/L AngⅡdistinctly increased the protein and mRNA expression of E-sel and MCP-1 in HUVECs(P＜0.01);Ang-(1-7)(1000 nmol/L) decreased the protein and mRNA expression of E-sel and MCP-1(P＜0.01).3.Compared with the AngⅡgroup,Ang-(1-7) inhibited synthesis of mRNA and protein as well as the secretion of cultured HUVECs induced by AngⅡin a dose-dependent manner (P＜0.01).4.The effects of Ang-(1-7) could be blocked by A-779.Conclusions1.AngⅡsignificantly induced the secretion of E-sel and MCP-1 in cultured HUVECs.2.Ang-(1-7) inhibited the secretion of E-sel and MCP-1.3.Ang-(1-7) inhibited the secretion of E-sel and MCP-1 in cultured HUVECs in a concentration dependent matter induced by AngⅡ.4.The inhibition effect of Ang-(1-7) maybe through the specific receptor.