Mulorz Joscha, Ibing Wiebke, Cappallo Melanie, Braß Sönke Maximilian, Takeuchi Kiku, Raaz Uwe, Schellinger Isabel Nahal, Krott Kim Jürgen, Schelzig Hubert, Aubin Hug, Oberhuber Alexander, Elvers Margitta, Wagenhäuser Markus Udo
Clinic for Vascular and Endovascular Surgery, Medical Faculty and University Hospital Duesseldorf, Heinrich-Heine-University, 40225 Duesseldorf, Germany.
Clinic for Cardiac Surgery, Medical Faculty and University Hospital Duesseldorf, Heinrich-Heine-University, 40225 Duesseldorf, Germany.
J Clin Med. 2023 Jul 28;12(15):4967. doi: 10.3390/jcm12154967.
(1) Background: Acute aortic dissection (AAD) is caused by an endothelial entry tear followed by intimomedial delamination of the outer layers of the vessel wall. The established risk factors include hypertension and smoking. Another rising candidate risk factor is excessive alcohol consumption. This experimental study explores the effects of nicotine (Nic), angiotensin II (Ang II), and ethanol (EtOH) on human aortic endothelial cells (hAoEC). (2) Methods: HAoECs were exposed to Nic, Ang II, and EtOH at different dose levels. Cell migration was studied using the scratch assay and live-cell imaging. The metabolic viability and permeability capacity was investigated using the water-soluble tetrazolium (WST)-1 assay and an in vitro vascular permeability assay. Cell adherence was studied by utilizing the hanging drop assay. The transcriptional and protein level changes were analyzed by RT-qPCR, Western blotting and immunohistochemistry for major junctional complexing proteins. (3) Results: We observed reduced metabolic viability following Ang II and EtOH exposure vs. control. Further, cell adherence was enhanced by EtOH exposure prior to trituration and by all risk factors after trituration, which correlated with the increased gene and protein expression of VE-cadherin upon EtOH exposure. The cell migration capacity was reduced upon EtOH exposure vs. controls. (4) Conclusion: Marked functional changes were observed upon exposure to established and potential risk factors for AAD development in hAoECs. Our findings advocate for an enhanced mechanical rigidity in hAoECs in response to the three substances studied, which in turn might increase endothelial rigidity, suggesting a novel mechanism for developing an endothelial entry tear due to reduced deformability in response to increased shear and pulsatile stress.
（1）背景：急性主动脉夹层（AAD）由内皮入口撕裂引发，随后血管壁外层发生内膜中层剥离。已确定的风险因素包括高血压和吸烟。另一个逐渐增加的潜在风险因素是过量饮酒。本实验研究探讨尼古丁（Nic）、血管紧张素II（Ang II）和乙醇（EtOH）对人主动脉内皮细胞（hAoEC）的影响。（2）方法：将hAoECs暴露于不同剂量水平的Nic、Ang II和EtOH。使用划痕试验和活细胞成像研究细胞迁移。使用水溶性四唑盐（WST）-1试验和体外血管通透性试验研究代谢活力和通透性。利用悬滴试验研究细胞黏附。通过RT-qPCR、蛋白质印迹法和免疫组织化学分析主要连接复合蛋白的转录和蛋白质水平变化。（3）结果：我们观察到，与对照组相比，Ang II和EtOH暴露后代谢活力降低。此外，研磨前EtOH暴露可增强细胞黏附，研磨后所有风险因素均可增强细胞黏附，这与EtOH暴露后VE-钙黏蛋白基因和蛋白表达增加相关。与对照组相比，EtOH暴露后细胞迁移能力降低。（4）结论：在hAoECs中，暴露于已确定的和潜在的AAD发生风险因素后观察到明显的功能变化。我们的研究结果表明，hAoECs对所研究的三种物质反应时机械刚性增强，这反过来可能增加内皮刚性，提示一种新机制，即由于对增加的剪切力和脉动应力的反应中变形性降低而导致内皮入口撕裂。
