Microvesicles derived from hypoxia/reoxYgenation-treated human umbilical vein endothellal cells impair relaxation of rat thoracic aortic rings  

作　　者：Shao-xun WANG Qi ZHANG Man SHANG Su WEI Miao LIU Yi-lu WANG Meng-xiao ZHANG Yan-na WU Ming-lin LIU Jun-qiu SONG Yan-xia LIU 

机构地区：[1]Department of Pharmacology,School of Basic Medical Sciences,Tianjin Medical University [2]Section of Endocrinology,Department of Medicine,Temple University School of Medicine [3]Department of Dermatology,Perelman School of Medicine,University of Pennsylvania

出　　处：《中国应用生理学杂志》2014年第6期560-566,共7页Chinese Journal of Applied Physiology

基　　金：supported by the Specialized Research Fund for the Doctoral Program of Higher Education of China(20101202110005);the Natural Science Foundation of Tianjin(11JCZDJC18300);the Research Foundation of Tianjin Municipal Education Commission(20110106);the National Key Basic Research Program of China(973 Program, 2011CB933100)

摘　　要：Objective To investigate the effects of microvesicles(MVs) derived from hypoxia/reoxygenation(H/R)-treated human umbilical vein endothelial cells(HUVECs) on endothelium-dependent relaxation of rat thoracic aortic rings.Methods H/R injury model was established to induce HUVECs to release H/R-EMVs.H/R-EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium.H/R-EMVs were characterized using 1 urn latex beads and anti-PE-CD144 by flow cytometry.Thoracic aortic rings of rats were incubated with 2.5,5,10,20 μg/ml H/R-EMVs derived from H/R-treated HUVECs for 4 hours,and their endothelium-dependent relaxation in response to acetylcholine(ACh) or endothelium-independent relaxation in response to sodium nitroprusside(SNP) was recorded in vitro.The nitric oxide(NO) production of ACh-treated thoracic aortic rings of rats was measured using Griess reagent.The expression of endothelial NO synthase(eNOS) and phosphorylated eNOS(p-eNOS,Ser-1177) in the thoracic aortic rings of rats was detected by Western blotting.Furthermore,the levels of SOD and MDA in H/R-EMVs-treated thoracic aortic rings of rats were measured using SOD and MDA kit.Results H/R-EMVs were induced by H/R-treated HUVECs and isolated by ultracentrifugation.The membrane vesicles(< 1 urn) induced by H/R were CD144 positive.ACh-induced relaxation and NO production of rat thoracic aortic rings were impaired by H/R-EMVs treatment in a concentration-dependent manner(P<0.05,P<0.01).The expression of total eNOS(t-eNOS)was not affected by H/R-EMVs.However,the expression of p-eNOS decreased after treated with H/R-EMVs.The activity of SOD decreased and the level of MDA increased in H/R-EMVs treated rat thoracic aortic rings(P<0.01).Conclusion ACh induced endothelium-dependent relaxation of thoracic aortic rings of rats was impaired by H/R-EMVs in a concentration-dependent manner.The mechanisms included a decrease in NO production,p-eNOS expression and an increase in oxidative stress.Objective To investigate the effects of microvesicles（MVs） derived from hypoxia/reoxygenation（H/R）-treated human umbilical vein endothelial cells（HUVECs） on endothelium-dependent relaxation of rat thoracic aortic rings.Methods H/R injury model was established to induce HUVECs to release H/R-EMVs.H/R-EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium.H/R-EMVs were characterized using 1 urn latex beads and anti-PE-CD144 by flow cytometry.Thoracic aortic rings of rats were incubated with 2.5,5,10,20 μg/ml H/R-EMVs derived from H/R-treated HUVECs for 4 hours,and their endothelium-dependent relaxation in response to acetylcholine（ACh） or endothelium-independent relaxation in response to sodium nitroprusside（SNP） was recorded in vitro.The nitric oxide（NO） production of ACh-treated thoracic aortic rings of rats was measured using Griess reagent.The expression of endothelial NO synthase（eNOS） and phosphorylated eNOS（p-eNOS,Ser-1177） in the thoracic aortic rings of rats was detected by Western blotting.Furthermore,the levels of SOD and MDA in H/R-EMVs-treated thoracic aortic rings of rats were measured using SOD and MDA kit.Results H/R-EMVs were induced by H/R-treated HUVECs and isolated by ultracentrifugation.The membrane vesicles（〈 1 urn） induced by H/R were CD144 positive.ACh-induced relaxation and NO production of rat thoracic aortic rings were impaired by H/R-EMVs treatment in a concentration-dependent manner（P〈0.05,P〈0.01）.The expression of total eNOS（t-eNOS）was not affected by H/R-EMVs.However,the expression of p-eNOS decreased after treated with H/R-EMVs.The activity of SOD decreased and the level of MDA increased in H/R-EMVs treated rat thoracic aortic rings（P〈0.01）.Conclusion ACh induced endothelium-dependent relaxation of thoracic aortic rings of rats was impaired by H/R-EMVs in a concentration-dependent manner.The mechanisms included a decrease in NO production,p-eNOS expression and an increase in oxidative stress.

关 键 词：人脐静脉内皮细胞 主动脉 大鼠 细胞来源 微泡 复氧 缺氧 浓度依赖性 

分 类 号：Q26[生物学—细胞生物学] TS201.22[轻工技术与工程—食品科学]