Use of a Mouse Model and Human Umbilical Vein Endothelial Cells to Investigate the Effect of Arsenic Exposure on Vascular Endothelial Function and the Associated Role of Calpains
Use of a Mouse Model and Human Umbilical Vein Endothelial Cells to Investigate the Effect of Arsenic Exposure on Vascular Endothelial Function and the Associated Role of Calpains
复制标题
使用小鼠模型和人脐静脉内皮细胞研究砷暴露对血管内皮功能的影响以及钙蛋白酶的相关作用
DOI：
10.1289/ehp4538
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发表时间：
2019-07-01
影响因子：
10.4
通讯作者：
Huang, Zhi
中科院分区：
文献类型：
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作者：
Cai, Zhihui;Zhang, Yanying;Huang, Zhi
背景：砷(As)是一种众所周知的环境污染物。众所周知，长期暴露于AS会增加心血管疾病的风险，包括动脉粥样硬化、高血压、糖尿病和中风。然而，AS导致涉及内皮完整性和通透性的血管功能障碍的详细机制尚不清楚。目的：我们的目标是调查暴露于AS是如何导致内皮功能障碍的。方法：采用小鼠血管内皮细胞单层培养模型，观察三氧化二砷(ATO)对血管内皮细胞功能障碍的影响，并探讨其作用机制。结果：与对照组相比，慢性染砷组小鼠对伊文思蓝染料和异硫氰酸荧光素标记的牛血清白蛋白(FITC-BSA)具有更强的血管通透性。此外，经ATO(0.13μM AS)处理的内皮单层显示出更大的细胞间隙和对低密度脂蛋白或迁移的THP-1细胞的通透性。此外，经ATO处理的主动脉和人脐静脉内皮细胞(HUVECs)中Calain-1(CAPN-1)的活性和蛋白水平显著升高。这些结果与ATO处理的结果一致，包括细胞内钙离子([Ca~(2+)]i)迅速升高和质膜CAPN-1水平升高。CAPN-1抑制剂(ALLM)而不是CAPN-2抑制剂(Z-LLY-FMK)可部分缓解ATO引起的内皮细胞功能障碍和血管内皮细胞钙粘附素(VE-cadherin)的蛋白降解紊乱。结论：本研究发现，在小鼠和HUVEC模型中，ATO通过增加[Ca~(2+)]i和CAPN-1向质膜的移位而导致CAPN-1的激活。这项研究还表明，抑制剂治疗可能在预防与AS暴露相关的血管内皮功能障碍方面发挥作用。本研究结果提示，AS诱导的内皮功能障碍涉及CAPN蛋白分解系统的过度激活。Https://doi.org/10.1289/EHP4538
Background: Arsenic (As) is a well-known environmental contaminant. Chronic exposure to As is known to increase the risk of cardiovascular diseases, including atherosclerosis, hypertension, diabetes, and stroke. However, the detailed mechanisms by which As causes vascular dysfunction involving endothelial integrity and permeability is unclear. Objectives: Our goal was to investigate how exposure to As leads to endothelial dysfunction. Methods: Arsenic trioxide (ATO) was used to investigate the effects and mechanisms by which exposure to As leads to endothelial dysfunction using a mouse model and cultured endothelial cell monolayers. Results: Compared with the controls, mice exposed chronically to As (10 ppb in drinking water supplied by ATO) exhibited greater vascular permeability to Evans blue dye and fluorescein isothiocyanate–labeled bovine serum albumin (FITC-BSA). In addition, endothelial monolayers treated with ATO (0.13μM As) exhibited greater intracellular gaps and permeability to low-density lipoprotein or transmigrating THP-1 cells. Furthermore, activity and protein levels of calpain-1 (CAPN-1) were significantly higher in aortas and human umbilical vein endothelial cells (HUVECs) treated with ATO. These results were consistent with effects of ATO treatment and included a rapid increase of intracellular calcium ([Ca2+]i) and higher levels of CAPN-1 in the plasma membrane. Endothelial cell dysfunction and the proteolytic disorganization of vascular endothelial cadherin (VE-cadherin) in HUVECs in response to ATO were partially mitigated by treatment with a CAPN-1 inhibitor (ALLM) but not a CAPN-2 inhibitor (Z-LLY-FMK). Conclusions: This study found that in mice and HUVEC models, exposure to ATO led to CAPN-1 activation by increasing [Ca2+]i and CAPN-1 translocation to the plasma membrane. The study also suggested that inhibitor treatment may have a role in preventing the vascular endothelial dysfunction associated with As exposure. The findings presented herein suggest that As-induced endothelial dysfunction involves the hyperactivation of the CAPN proteolytic system. https://doi.org/10.1289/EHP4538