The present study aimed to investigate the impacts and underlying mechanisms of 14,15-DHETs on eNOS and vascular endothelial functions. Bovine aortic endothelial cells (BAECs) were treated with various concentrations of 14, 15-DHET. The expressions of eNOS protein and mRNA were observed at different time points. The eNOS expression and phosphorylation were subsequently detected administered with 8,9-DHET, 11,12-DHET, and 14,15-DHET, respectively. Meanwhile, 14,15-DHET action on tube formation was observed in human umbilical vein endothelial cells (HUVECs). Finally, the aorta of male C57BL/6 mice was injected with 14,15-DHET via the tail vein. The impacts of 14,15-DHET (0.4 mg/kg body weight) on the expressions of eNOS protein and mRNA and endothelium-dependent vasodilation (EDV) were detected following 24 h. The expression of eNOS was greatly improved with the 14,15-DHET treatment compared with the BAECs, and eNOS phosphorylation sites at Ser1179, Ser635, and Thr497 were elevated. However, no statistically significant difference was revealed on total eNOS among the 8,9-DHET, 11,12-DHET, and 14,15-DHET treatment groups. Based on the upregulation of eNOS protein, eNOS mRNA levels were increased in BAECs and thoracic aorta of the male C57BL/6 mice treated with 14,15-DHET, suggesting that transcriptional activation was achieved in vascular eNOS. Moreover, 14,15-DHET enhanced tube formation abilities in HUVECs and acetylcholine(ACh)-induced EDV. These findings indicated that 14,15-DHET could improve the vascular endothelial diastolic functions of male C57BL/6 mice, and enhance the ability of tube formation, which might be related to the increase of eNOS expression.

本研究旨在调查 14,15-DHETs 对 eNOS 和血管内皮功能的影响和潜在机制。牛主动脉内皮细胞 (BAEC) 用不同浓度的 14, 15-DHET 处理。在不同时间点观察eNOS蛋白和mRNA的表达。随后分别与 8,9-DHET、11,12-DHET 和 14,15-DHET 一起检测 eNOS 表达和磷酸化。同时，在人脐静脉内皮细胞 (HUVEC) 中观察到 14,15-DHET 对管形成的作用。最后，通过尾静脉向雄性 C57BL/6 小鼠的主动脉注射 14,15-DHET。24 小时后检测到 14,15-DHET（0.4 mg/kg 体重）对 eNOS 蛋白和 mRNA 表达以及内皮依赖性血管舒张 (EDV) 的影响。与 BAEC 相比，14,15-DHET 处理后 eNOS 的表达大大提高，并且 Ser1179、Ser635 和 Thr497 处的 eNOS 磷酸化位点升高。然而，在 8,9-DHET、11,12-DHET 和 14,15-DHET 治疗组之间的总 eNOS 没有显示出统计学上的显着差异。基于 eNOS 蛋白的上调，用 14,15-DHET 处理的雄性 C57BL/6 小鼠的 BAEC 和胸主动脉中的 eNOS mRNA 水平增加，表明在血管 eNOS 中实现了转录激活。此外，14,15-DHET 增强了 HUVEC 和乙酰胆碱 (ACh) 诱导的 EDV 中的管形成能力。这些发现表明14,15-DHET可以改善雄性C57BL/6小鼠血管内皮舒张功能，增强管形成能力，