The Influence Of Angiotensin-(1-7) On The Secretion Of ICAM-1 And VCAM-1 In Cultured Human Umbilical Vein Cells Induced By AngiotensinⅡ

Background & ObjectivesRecent studies suggest that atherosclerosis is a chronic inflammatory disease.Endothelial cell damage and Monocyte adhesion to endothelial cells are important initial events at the onset of atherosclerosis.Intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)are critical adhesion molecules existing on vascular endothelial cells, which promote monocyte adhering to endothelial cells so as to initiate atherosclerosis.And according to the researches,AngiotensinⅡ(AngⅡ)is an important active factor in renin-angiotensin system(RAS).AngⅡcan stimulate the increase of ICAM-1 and VCAM-1, which plays a cruial role in atherosclerosis.Angiotensin-(1-7)[Ang-(1-7)]is a new member of the renin-angiotensin system.It can prevent AngⅡeffects,for example,having antihypertensive, antiproliferative and diuresis effects.However,it remains unknown whether Ang-(1-7)can prevent the increase of ICAM-1 and VCAM-1 in human umbilical vein Cells(HUVEC)induced by AngⅡ.In this experiment,we explore the influence of angiotensin-(1-7)[Ang-(1-7)]on the secretion of ICAM and VCAM in cultured HUVEC Induced by AngⅡby cultivating the umbilical vein Cells and taking the measures of detecting ICAM-1 and VCAM-1 protein and mRNA expression by ELISA and RT-PCR.And we aim to produce the other new way by stating the prevent effect of Ang-(1-7)Induced by AngⅡon the inflammation,and provide the theory basis for the clinical cardiovascular diseases.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)10000mol/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 1000nmol/L for 30 minute,then pretreated with the final concentration Ang-(1-7)1000nmol/L for 30 minute,finally stimulated with the final concentration AngⅡ100nmol/L. 3.The HUVECs ICAM-1 and VCAM-1 protein expression were measured by enzyme linked immunosorbent assay(ELISA),and ICAM-1 and VCAM-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 ICAM-1 and VCAM-1 in HUVECs(P＜0.05);Ang-(1-7)(1000 nmol/L) decreased the protein and mRNA expression of ICAM-1 and VCAM-1(P＜0.05).When the concentration of Ang-(1-7)was up to 1000nmol/L,the expression of ICAM-1 and VCAM-1 were more than control,but was not statistically significant.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＜0.05).4.The effects of Ang-(1-7)could be blocked by A-779.Conclusions1.AngⅡsignificantly induced the secretion of ICAM-1 and VCAM-1 in cultured HUVECs.2.Ang-(1-7)inhibited the secretion of ICAM-1 and VCAM-1.3.Ang-(1-7)inhibited the secretion of ICAM-1 and VCAM-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.