Department of Respiratory Medicine , The First Affiliated Hospital of Wenzhou Medical College, China.
Free Radic Res. 2012 Dec;46(12):1437-45. doi: 10.3109/10715762.2012.724532. Epub 2012 Sep 27.
The objective was to investigate the molecular mechanism of mitochondrial reactive oxygen species (ROS) signaling regulation of pulmonary artery endothelial cell (HPAEC) secretion in the condition of oxidative stress. Acrolein (40 μM) induced HPAEC mitochondrial generation of ROS, rotenone (2 μmol/L) blocked mitochondrial respiratory chain complex I, cesium chloride (CsCl, 40 mmol/L)blocked K(+)channels, and saline (0.9 g/dl) were used as control. The generations of NOS, ET-1 and VEGF were determined with ELISA in the condition of different treatment reagents namely acrolein, acrolein plus rotenone, acrolein plus CsCl and saline. In the different reagent treatment of HPAECs, acrolein increased mitochondrial ROS, membrane potential, Kv1.5 mRNA and protein expression, intracellular calcium and the generation of NOS (determining NO production), ET-1 and VEGF, and those were reduced by rotenone. CsCl decreased the increment of membrane potential, the elevation of intracellular calcium and the upregulation of NOS, E-1 and VEGF expressions, which were induced by acrolein. The present study demonstrated that mitochondrial ROS-K(+)channel regulated HPAEC secretion of NO, ET-1 and VEGF in the condition of oxidative stress. Kv1.5 channel may be an important component of ROS-K+ channel signaling pathway, and intracellular calcium contributed to mitochondrial ROS-K(+) channel signaling modulation of HPAEC secretion.
目的在于研究氧化应激条件下，线粒体活性氧（ROS）信号调节肺动脉内皮细胞（HPAEC）分泌的分子机制。丙烯醛（40 μM）诱导 HPAEC 线粒体产生 ROS，鱼藤酮（2 μmol/L）阻断线粒体呼吸链复合物 I，氯化铯（CsCl，40 mmol/L）阻断 K+通道，生理盐水（0.9 g/dl）用作对照。在不同处理试剂（丙烯醛、丙烯醛加鱼藤酮、丙烯醛加 CsCl 和生理盐水）的条件下，用 ELISA 法测定 NOS、ET-1 和 VEGF 的生成。在 HPAECs 的不同试剂处理中，丙烯醛增加了线粒体 ROS、膜电位、Kv1.5 mRNA 和蛋白表达、细胞内钙和 NOS（测定 NO 生成）、ET-1 和 VEGF 的生成，而鱼藤酮则降低了这些生成。CsCl 降低了膜电位的增加、细胞内钙的升高和由丙烯醛诱导的 NOS、E-1 和 VEGF 表达的上调。本研究表明，线粒体 ROS-K+通道在氧化应激条件下调节 HPAEC 分泌 NO、ET-1 和 VEGF。Kv1.5 通道可能是 ROS-K+通道信号通路的重要组成部分，细胞内钙有助于线粒体 ROS-K+通道信号调节 HPAEC 分泌。
