Division of Clinical Pharmacology (J.K., N.d.l.V., V.A., D.G.H., D.M.P.), Department of Medicine, Vanderbilt University Medical Center.
Department of Biomedical Engineering, Vanderbilt University (E.M.H.).
Hypertension. 2022 Aug;79(8):1644-1655. doi: 10.1161/HYPERTENSIONAHA.122.19305. Epub 2022 Jun 10.
IsoLGs (isolevuglandins) are electrophilic products of lipid peroxidation formed in the presence of reactive oxygen species. IsoLGs contribute to hypertension by an unknown mechanism. Studies have shown that reactive oxygen species production drives the formation of neutrophil extracellular traps (NETs) and that NETs accumulate within the aorta and kidneys of patients with hypertension. The purpose of this study was to determine the role of isoLGs in neutrophil migration and NET formation (NETosis) in hypertension.
Mice were treated with Ang II (angiotensin II) and the specific isoLG scavenger 2-hydroxybenzylamine and examined for tissue neutrophil and NET accumulation by single-cell sequencing and flow cytometry. Isolated human neutrophils were studied to determine the role of isoLGs in NETosis and neutrophil chromatin expansion by immunofluorescence and live cell confocal microscopy.
Single-cell sequencing performed on sham, Ang II, and Ang II+2-hydroxybenzylamine treated mice revealed neutrophils as a primary target of 2-hydroxybenzylamine. Peripheral neutrophil migration, aortic NET accumulation, and renal NET accumulation is blocked with 2-hydroxybenzylamine treatment. In isolated human neutrophils, isoLGs accumulate during NETosis and scavenging of isoLGs prevents NETosis. IsoLGs drive neutrophil chromatin expansion during NETosis and disrupt nucleosome structure.
These observations identified a critical role of isoLGs in neutrophil migration and NETosis in hypertension and provide a potential therapy for NET-associated diseases including hypertension and associated end organ damage.
异莱格（isolevuglandins）是在活性氧物质存在下形成的脂质过氧化的亲电产物。异莱格通过未知机制导致高血压。研究表明，活性氧物质的产生会导致中性粒细胞胞外诱捕网（NETs）的形成，而 NETs 在高血压患者的主动脉和肾脏中积聚。本研究旨在确定异莱格在高血压中性粒细胞迁移和 NET 形成（NETosis）中的作用。
用血管紧张素 II（angiotensin II）和特异性异莱格清除剂 2-羟苯甲胺处理小鼠，通过单细胞测序和流式细胞术检测组织中性粒细胞和 NET 积聚。分离人中性粒细胞，通过免疫荧光和活细胞共聚焦显微镜确定异莱格在 NETosis 和中性粒细胞染色质扩张中的作用。
对假手术、Ang II 和 Ang II+2-羟苯甲胺处理的小鼠进行单细胞测序显示，中性粒细胞是 2-羟苯甲胺的主要靶标。用 2-羟苯甲胺处理可阻止外周中性粒细胞迁移、主动脉 NET 积聚和肾 NET 积聚。在分离的人中性粒细胞中，在 NETosis 过程中异莱格积聚，清除异莱格可防止 NETosis。异莱格在 NETosis 过程中驱动中性粒细胞染色质扩张并破坏核小体结构。
这些观察结果确定了异莱格在高血压中性粒细胞迁移和 NETosis 中的关键作用，并为包括高血压和相关终末器官损伤在内的 NET 相关疾病提供了一种潜在的治疗方法。
