Transient Receptor Potential Melastatin 4 (TRPM4) Contributes to High Salt Diet-Mediated Early-Stage Endothelial Injury
Transient Receptor Potential Melastatin 4 (TRPM4) Contributes to High Salt Diet-Mediated Early-Stage Endothelial Injury
复制标题
瞬时受体电位褪黑激素 4 (TRPM4) 会导致高盐饮食介导的早期内皮损伤。
DOI：
10.1159/000459695
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Zhang, Zhi-Ren
中科院分区：
文献类型：
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作者：
Ding, Xiao-Qing;Ban, Tao;Zhang, Zhi-Ren
背景/目的：本研究旨在探讨瞬时受体潜在蛋白4(TRPM4)通道在高盐饮食(HSD)诱导的内皮损伤中的作用。方法：用Western blotting和免疫荧光技术检测给予HSD的Dahl盐敏感(SS)大鼠肠系膜上皮细胞TRPM4的表达。采用四甲基偶氮唑盐比色法、原位末端标记法和Transwell法分别检测人脐静脉内皮细胞(HUVECs)的存活率、细胞凋亡率和细胞迁移。采用酶联免疫吸附试验检测细胞间黏附分子-1(ICAM-1)、血管细胞黏附蛋白-1(VCAM-1)和E-选择素的浓度。用膜通透性的活性氧(ROS)敏感的荧光探针Carboxy-H2DCFDA检测细胞内ROS水平。结果：TRPM4主要在肠系膜动脉内皮细胞质膜附近表达，给予HSD的SS高血压大鼠其表达水平增加。外源性过氧化氢(H_2O_2)和醛固酮作用于培养的人脐静脉内皮细胞后，其蛋白表达明显上调。当两种药物以浓度依赖的方式处理时，细胞存活率下降，这可以被TRPM4的特异性抑制剂9-菲罗酚部分逆转。外源性过氧化氢诱导细胞凋亡，促进细胞迁移，增加黏附分子的释放，包括ICAM-1、VCAM-1和E-选择素，这些都在9-菲咯醇处理后显著减弱。醛固酮和H_2O_2诱导细胞内ROS的积累，并被9-菲咯啉显著抑制，提示氧化应激是醛固酮诱导内皮损伤的机制之一。结论：鉴于高血压患者体内存在氧化应激和循环中高水平的醛固酮，我们认为TRPM4在血管内皮细胞中的表达上调可能参与了这些刺激引起的内皮损伤。(C)2017年作者(S)，S.Karger AG出版，
Background/Aims: The present study investigated whether the transient receptor potential melastatin 4 (TRPM4) channel plays a role in high salt diet (HSD)-induced endothelial injuries. Methods: Western blotting and immunofluorescence were used to examine TRPM4 expression in the mesenteric endothelium of Dahl salt-sensitive ( SS) rats fed a HSD. The MTT, TUNEL, and transwell assays were used to evaluate the cell viability, cell apoptosis, and cell migration, respectively, of human umbilical vein endothelial cells (HUVECs). Enzyme-linked immunosorbent assays were used to determine the concentrations of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion protein 1 (VCAM-1), and E-selectin. Carboxy-H2DCFDA, a membrane-permeable reactive oxygen species (ROS)-sensitive fluorescent probe, was used to detect intracellular ROS levels. Results: TRPM4 was mainly expressed near the plasma membrane of mesenteric artery endothelial cells, and its expression level increased in SS hypertensive rats fed a HSD. Its protein expression was significantly upregulated upon treatment with exogenous hydrogen peroxide (H2O2) and aldosterone in cultured HUVECs. Cell viability decreased upon treatment with both agents in a concentration-dependent manner, which could be partially reversed by 9-phenanthrol, a specific TRPM4 inhibitor. Exogenous H2O2 induced apoptosis, enhanced cell migration, and increased the release of adhesion molecules, including ICAM-1, VCAM-1, and E-selectin, all of which were significantly attenuated upon treatment with 9-phenanthrol. Aldosterone and H2O2 induced the accumulation of intracellular ROS, which was significantly inhibited by 9-phenanthrol, suggesting that oxidative stress is one of the mechanisms underlying aldosterone-induced endothelial injury. Conclusions: Given the fact that oxidative stress and high levels of circulating aldosterone are present in hypertensive patients, we suggest that the upregulation of TRPM4 in the vascular endothelium may be involved in endothelial injuries caused by these stimuli. (C) 2017 The Author(s) Published by S. Karger AG,