Yang Jining, Zhou Xi, Zeng Xianglong, Hu Ou, Yi Long, Mi Mantian
Research Center for Nutrition and Food Safety, Chongqing Key Laboratory of Nutrition and Food Safety, Institute of Military Preventive Medicine, Third Military Medical University, 30th Gaotanyan Main Street, Shapingba District, Chongqing, 400038 People's Republic of China.
Nutr Metab (Lond). 2019 Jan 30;16:9. doi: 10.1186/s12986-019-0338-7. eCollection 2019.
BACKGROUND/AIMS: Oxidative stress-induced damage in endothelial cells is a crucial initiator of atherosclerosis (AS), which is highly related to excessive reactive oxygen species (ROS) and mitochondrial dynamics. Resveratrol (RSV) exerts beneficial effects against endothelial oxidative injury, while the underlying mechanisms have not been fully elucidated. Thus, we aimed to explore the role of mitochondria dynamics during the anti-oxidative activities of RSV in palmitic acid (PA)-stimulated human umbilical vein endothelial cells (HUVECs) and to verify whether tyrosyl transfer- RNA synthetase (TyrRS) and poly (ADP-ribose) polymerase 1 (PARP1) are targeted during this process.
HUVECs were exposed to 200 μM of PA for 16 h before treated with 10 μM of RSV for 8 h. Cell viability was detected using Cell counting kit-8 (CCK-8) assay. The intracellular ROS level and mitochondria membrane potential (MMP) were measured using microplate reader and flow cytometry. The malondialdehyde and superoxide dismutase were measured using the microplate reader. The mitochondrial morphology and fusion process was observed under transmission electron microscopy and confocal microscopy. TyrRS and PARP1 were knocked down with the specific small interference RNAs (siRNA), and the protein expressions of TyrRS, PARP1, and mitochondrial fusion proteins (MFN1, MFN2, and OPA1) were measured by western blot.
RSV treatment suppressed the PA-induced injuries in HUVECs, including the damage to cell viability, oxidative stress, and loss of MMP. Additionally, RSV improved the protein levels of MFN1, MFN2, and OPA1 as well as inhibited the PA-induced fragmentation of mitochondria. However, the effects of RSV on oxidative stress and mitochondrial fusion were abolished by the pretreatment of siRNAs of TyrRS and PARP1, indicating that these effects of RSV were dependent on the TyrRS-PARP1 pathway.
RSV attenuated endothelial oxidative injury by regulating mitochondrial fusion via TyrRS-PARP1 signaling pathway.
背景/目的：内皮细胞中氧化应激诱导的损伤是动脉粥样硬化（AS）的关键启动因素，这与过量的活性氧（ROS）和线粒体动力学高度相关。白藜芦醇（RSV）对内皮氧化损伤具有有益作用，但其潜在机制尚未完全阐明。因此，我们旨在探讨线粒体动力学在RSV对棕榈酸（PA）刺激的人脐静脉内皮细胞（HUVECs）抗氧化活性中的作用，并验证在此过程中酪氨酰转移RNA合成酶（TyrRS）和聚（ADP - 核糖）聚合酶1（PARP1）是否为作用靶点。
HUVECs在暴露于200μM PA 16小时后，用10μM RSV处理8小时。使用细胞计数试剂盒 - 8（CCK - 8）检测细胞活力。使用酶标仪和流式细胞仪测量细胞内ROS水平和线粒体膜电位（MMP）。使用酶标仪测量丙二醛和超氧化物歧化酶。在透射电子显微镜和共聚焦显微镜下观察线粒体形态和融合过程。用特异性小干扰RNA（siRNA）敲低TyrRS和PARP1，并通过蛋白质印迹法测量TyrRS、PARP1和线粒体融合蛋白（MFN1、MFN2和OPA1）的蛋白表达。
RSV处理抑制了PA诱导的HUVECs损伤，包括对细胞活力的损害、氧化应激和MMP的丧失。此外，RSV提高了MFN1、MFN2和OPA1的蛋白水平，并抑制了PA诱导的线粒体碎片化。然而，TyrRS和PARP1的siRNAs预处理消除了RSV对氧化应激和线粒体融合的影响，表明RSV的这些作用依赖于TyrRS - PARP1途径。
RSV通过TyrRS - PARP1信号通路调节线粒体融合，减轻内皮氧化损伤。
