Iptakalim ameliorates relaxation to acetylcholine in thoracic aortic rings impaired by microvesicles derived from hypoxia/reoxygenation-treated HUVECs

作     者：Kun-wei ZHANG Shao-xun WANG Ye-yi LI Su WEI Man SHANG Chao LIU Miao LIU Yi-lu WANG Qian ZHU Yan-na WU Jun-qiu SONG Yan-xia LIU Kun-wei ZHANG;Shao-xun WANG;Ye-yi LI;Su WEI;Man SHANG;Chao LIU;Miao LIU

作者机构：Department of PharmacologySchool of Basic Medical SciencesTianjin Medical University 

出 版 物：《中国应用生理学杂志》 (Chinese Journal of Applied Physiology)

年 卷 期：2016年第32卷第6期

页      面：481-486页

核心收录：

学科分类：0710[理学-生物学] 0832[工学-食品科学与工程（可授工学、农学学位）] 1002[医学-临床医学] 07[理学] 08[工学] 071009[理学-细胞生物学] 09[农学] 0901[农学-作物学] 083201[工学-食品科学] 090102[农学-作物遗传育种] 

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) 

主　　题：人脐静脉内皮细胞 主动脉 丝氨酸/苏氨酸激酶 复氧 缺氧 微泡 一氧化氮合酶 透射电子显微镜 

摘      要：Objective: To investigate the effect of Iptakalim(Ipt) preventing injury of endothelial microvesicles(EMVs) derived from hypoxia/reoxygenation(H/R)-treated HUVECs on the relaxation of rat thoracic aortic rings and explore the underlying mechanism. Methods: H/R injury model was established to release H/R-EMVs from HUVECs. H/R-EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium. H/R-EMVs were characterized by using Transmission Electron Microscope(TEM). Thoracic aortic rings of rats were incubated with 10^(-7)-10^(-3 )mol/L Ipt and co-cultured with 10 μg/ml H/R-EMVs for 4 hours, and their endothelium- dependent relaxation in response to acetylcholine(ACh) was recorded in vitro. The nitric oxide(NO) production of ACh-treated rat thoracic aortic rings was measured by using Griess reagent. The expression of endothelial NO synthase(e NOS), phosphorylated e NOS(p-e NOS, Ser-1177), serine/threonine kinas(Akt) and phosphorylated Akt(p-Akt, Ser-473) in the thoracic aortic rings of rats was detected by Western blotting. Results: H/R-EMVs were induced by H/R-treated HUVECs and isolated by ultracentrifugation. The isolated H/R-EMVs subjected to TEM revealed small, rounded vesicles(100–1 000 nm) surrounded by a membrane. H/R-EMVs impaired relaxation induced by ACh of rat thoracic aortic rings significantly. Compared with H/R-EMVs treatment individually, relaxation and NO production of rat thoracic aortic rings were increased by Ipt treatment in a concentration-dependent manner(P0.05, P0.01). The expression of total e NOS(t-e NOS) and total Akt(t-Akt) was not affected by Ipt or H/R-EMVs. However, the expression of p-e NOS and p-Akt increased after treated with Ipt(P0.01). Conclusion: Based on H/R-EMVs treatment, ACh induced endothelium-dependent relaxation of rat thoracic aortic rings was ameliorated by Ipt in a concentration-dependent manner. The mechanisms involved the increase in NO production, p-e NOS and p-Akt expression.