Hypoxia reoxygenation (HR) injury perturbs structural and functional syncytium in lung tissues. It is commonly implicated in conditions such as stroke, lung transplant or severe pneumonia. In the present study, we investigated the cytoprotective action of 20-hydroxyeicosatetraenoic acid (20-HETE) on pulmonary vascular endothelial cells (PMVECs) under normoxic and hypoxic niche followed by HR. 20-HETE pretreatment showed a protective effect at a concentration of 1μM as there was a marked increase (20%) in the cell viability compared to control and HR groups. Pretreatment of 20-HETE in HR induced injury decreased ROS production dictated its antioxidant property. Similarly, SOD and ATP levels were also downregulated by 20-HETE pretreatment. Cell apoptosis was detected by TUNEL assay, Acridine orange, and procaspase-3 cleavage, caspase-3 activity assay, respectively. JC-1 mitochondrial membrane potential assay and protein expression pattern of BCL-2, and BAD phosphorylation status were examined. The results showed that HR induced significant increase of apoptotic PMVECs, while 20-HETE pretreatment attenuated the effects. Further, 20-HETE pretreatment activated PI3K/Akt and HIF-1α signaling pathway to exhibit its protective effects against HR-induced oxidative stress and apoptosis. Overall, the results concluded the potent antioxidant role of 20-HETE in aiding cytoprotection upon HR injury.

中文翻译：

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预防性补充20-HETE可通过抑制细胞凋亡来改善肺血管内皮细胞的缺氧/复氧损伤。

缺氧复氧（HR）损伤扰乱了肺组织的结构和功能合胞体。通常与中风，肺移植或严重肺炎等疾病有关。在本研究中，我们调查了20-羟基二十碳四烯酸（20-HETE）在常氧和低氧环境下，然后在HR下对肺血管内皮细胞（PMVEC）的细胞保护作用。20-HETE预处理在浓度为1μM时显示出保护作用，因为与对照组和HR组相比，细胞活力显着增加（20％）。HR损伤中20-HETE的预处理降低了ROS的产生，从而决定了其抗氧化性能。同样，20-HETE预处理也下调了SOD和ATP水平。通过TUNEL分析，A啶橙和procaspase-3裂解检测细胞凋亡，caspase-3活性测定。检查了JC-1线粒体膜电位测定和BCL-2的蛋白表达模式，以及BAD的磷酸化状态。结果表明，HR诱导凋亡PMVEC显着增加，而20-HETE预处理则减弱了这种作用。此外，20-HETE预处理激活了PI3K / Akt和HIF-1α信号通路，以显示其对HR诱导的氧化应激和细胞凋亡的保护作用。总体而言，该结果得出结论：20-HETE在HR损伤时有助于细胞保护的有效抗氧化剂作用。20-HETE预处理激活了PI3K / Akt和HIF-1α信号通路，以显示其对HR诱导的氧化应激和细胞凋亡的保护作用。总体而言，该结果得出结论：20-HETE在HR损伤时有助于细胞保护的有效抗氧化剂作用。20-HETE预处理激活了PI3K / Akt和HIF-1α信号通路，以显示其对HR诱导的氧化应激和细胞凋亡的保护作用。总体而言，该结果得出结论：20-HETE在HR损伤时有助于细胞保护的有效抗氧化剂作用。