Orai-vascular endothelial-cadherin signaling complex regulates high-glucose exposure-induced increased permeability of mouse aortic endothelial cells.
Orai-vascular endothelial-cadherin signaling complex regulates high-glucose exposure-induced increased permeability of mouse aortic endothelial cells.
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DOI：
10.1136/bmjdrc-2020-002085
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发表时间：
2021-04
影响因子：
4.1
通讯作者：
Du J
中科院分区：
文献类型：
--
作者：
Wei Y;Bai S;Yao Y;Hou W;Zhu J;Fang H;Du Y;He W;Shen B;Du J
糖尿病相关的内皮屏障功能损害可能与Ca 2+稳态紊乱有关。目的：研究小鼠主动脉内皮细胞（MAECs）中Orais-血管内皮细胞（VE）-钙粘蛋白信号复合物及其下游信号通路在糖尿病血管内皮损伤中的作用及分子机制。高糖（HG）和正常葡萄糖（NG）暴露7 d后，采用钙离子成像技术检测细胞钙库操纵的钙内流（SOCE）活性，Western blot检测高糖处理后Orais的表达水平。采用免疫荧光法观察HG暴露对细胞膜磷酸化（p）-VE-钙粘蛋白和VE-钙粘蛋白表达的影响。体外培养MAECs，观察HG诱导的跨内皮电阻。通过transwell渗透性测定法检测FD-20在单层主动脉内皮细胞中的渗透性。采用免疫共沉淀和免疫荧光技术检测Orais与VE-钙粘蛋白的相互作用。免疫组化法检测糖尿病小鼠主动脉内皮Orais、VE-钙粘素和p-VE-钙粘素的表达变化。(1)高糖培养7 d的MAECs Orais表达水平和SOCE活性显著升高。(2)高糖培养7 d的MAECs，细胞膜p-VE-cadherin/VE-cadherin比值增加，单层内皮细胞FD-20通透性增加，表明细胞间通透性增加。(3)Orais和VE-钙粘蛋白可以相互作用，并通过HG刺激增强相互作用的比率。(4)SOCE激活剂ATP可促进高糖培养的MAECs p-VE-cadherin的表达，而SOCE抑制剂BTP 2则可降低p-VE-cadherin的表达。(5)糖尿病小鼠主动脉内皮细胞p-VE-cadherin和Orais的表达显著增加。HG暴露刺激内皮细胞Orais表达增加，通过Orais-VE-cadherin复合物及一系列下游信号通路增加VE-cadherin磷酸化，导致内皮细胞连接破坏，动脉粥样硬化形成。
Diabetes-associated endothelial barrier function impairment might be linked to disturbances in Ca2+ homeostasis. To study the role and molecular mechanism of Orais–vascular endothelial (VE)-cadherin signaling complex and its downstream signaling pathway in diabetic endothelial injury using mouse aortic endothelial cells (MAECs). The activity of store-operated Ca2+ entry (SOCE) was detected by calcium imaging after 7 days of high-glucose (HG) or normal-glucose (NG) exposure, the expression levels of Orais after HG treatment was detected by western blot analysis. The effect of HG exposure on the expression of phosphorylated (p)-VE-cadherin and VE-cadherin on cell membrane was observed by immunofluorescence assay. HG-induced transendothelial electrical resistance was examined in vitro after MAECs were cultured in HG medium. FD-20 permeability was tested in monolayer aortic endothelial cells through transwell permeability assay. The interactions between Orais and VE-cadherin were detected by co-immunoprecipitation and immunofluorescence technologies. Immunohistochemical experiment was used to detect the expression changes of Orais, VE-cadherin and p-VE-cadherin in aortic endothelium of mice with diabetes. (1) The expression levels of Orais and activity of SOCE were significantly increased in MAECs cultured in HG for 7 days. (2) In MAECs cultured in HG for 7 days, the ratio of p-VE-cadherin to VE-cadherin expressed on the cell membrane and the FD-20 permeability in monolayer endothelial cells increased, indicating that intercellular permeability increased. (3) Orais and VE-cadherin can interact and enhance the interaction ratio through HG stimulation. (4) In MAECs cultured with HG, the SOCE activator ATP enhanced the expression level of p-VE-cadherin, and the SOCE inhibitor BTP2 decreased the expression level of p-VE-cadherin. (5) Significantly increased expression of p-VE-cadherin and Orais in the aortic endothelium of mice with diabetes. HG exposure stimulated increased expression of Orais in endothelial cells, and increased VE-cadherin phosphorylation through Orais–VE-cadherin complex and a series of downstream signaling pathways, resulting in disruption of endothelial cell junctions and initiation of atherosclerosis.
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影响因子：
5.3
作者：
Chen Y;Pitzer AL;Li X;Li PL;Wang L;Zhang Y
通讯作者：
Zhang Y
影响因子：
7.5
作者：
Harris ES;Nelson WJ
通讯作者：
Nelson WJ
影响因子：
3.3
作者：
Lander, Heather M.;Grant, Ashley M.;Peters, Clarence J.
通讯作者：
Peters, Clarence J.
DOI：
10.1073/pnas.94.12.6273
发表时间：
1997-06-10
影响因子：
11.1
作者：
Vittet, D;Buchou, T;Huber, P
通讯作者：
Huber, P
影响因子：
7.5
作者：
Huang, Yuan-Yuan;Li, Jing;Xia, Xian-Ming
通讯作者：
Xia, Xian-Ming