Central role of c-Src in NOX5- mediated redox signalling in vascular smooth muscle cells in human hypertension.
Central role of c-Src in NOX5- mediated redox signalling in vascular smooth muscle cells in human hypertension.
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DOI：
10.1093/cvr/cvab171
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发表时间：
2022-03-25
影响因子：
10.8
通讯作者：
Touyz RM
中科院分区：
文献类型：
--
作者：
Camargo LL;Montezano AC;Hussain M;Wang Y;Zou Z;Rios FJ;Neves KB;Alves-Lopes R;Awan FR;Guzik TJ;Jensen T;Hartley RC;Touyz RM
NOX 衍生的活性氧 (ROS) 是与高血压血管平滑肌细胞 (VSMC) 功能障碍有关的信号通路的介质。在 VSMC 调节中重要的众多氧化还原敏感激酶中，c-Src 是其中之一。然而，NOX/ROS 与 c-Src 的联系机制尚不清楚，特别是在高血压氧化应激的背景下。在这里，我们研究了 NOX 诱导的 VSMC 氧化应激在人类高血压中的作用，重点关注 NOX5，并探索了 c-Src 作为连接 NOX5-ROS 与潜在 VSMC 功能障碍的下游效应靶标的假定中间体。研究了血压正常（NT）和高血压（HT）受试者动脉的 VSMC。在 NOX5（蜂毒肽）和 Src（PP2）抑制剂存在和不存在的情况下评估 NOX1、2、4、5 表达、ROS 生成、信号分子的氧化/磷酸化以及肌动蛋白聚合和迁移。使用 NOX5 siRNA 和 p22phox-siRNA 方法下调 NOX5 和 p22phox 依赖性 NOX (NOX1–4)。作为完整血管的概念证明，通过肌电图在以 VSMC 特异性方式表达人 NOX5 的转基因小鼠中评估血管功能。在 HT VSMC 中，NOX5 上调，与信号分子的氧化应激、过度氧化（c-Src、过氧化还原蛋白、DJ-1）和过度磷酸化（c-Src、PKC、ERK1/2、MLC20）相关。 NOX5 siRNA 减少 NT 和 HT 受试者中 ROS 的产生。 NOX5 siRNA（而非 p22phox-siRNA）减弱了 HT VSMC 中的 c-Src 磷酸化。 NOX5 siRNA 降低 NT 和 HT 中 MLC20 和 FAK 的磷酸化。在 p22phox 沉默的 HT VSMC 中，Ang II 诱导的 MLC20 磷酸化增加，但效果被蜂毒肽和 PP2 阻断。 NOX5 和 c-Src 抑制减弱了 HT VSMC 中的肌动蛋白聚合和迁移。在 NOX5 转基因小鼠中，蜂毒肽和 PP2 降低了血管过度收缩。我们将 NOX5/ROS/c-Src 定义为人类 VSMC 中的新型前馈信号网络。该系统在高血压中的放大会导致 VSMC 功能障碍。抑制 NOX5/ROS/c-Src 通路可能会改善高血压相关的血管损伤。
NOX-derived reactive oxygen species (ROS) are mediators of signalling pathways implicated in vascular smooth muscle cell (VSMC) dysfunction in hypertension. Among the numerous redox-sensitive kinases important in VSMC regulation is c-Src. However, mechanisms linking NOX/ROS to c-Src are unclear, especially in the context of oxidative stress in hypertension. Here, we investigated the role of NOX-induced oxidative stress in VSMCs in human hypertension focusing on NOX5, and explored c-Src, as a putative intermediate connecting NOX5-ROS to downstream effector targets underlying VSMC dysfunction. VSMC from arteries from normotensive (NT) and hypertensive (HT) subjects were studied. NOX1,2,4,5 expression, ROS generation, oxidation/phosphorylation of signalling molecules, and actin polymerization and migration were assessed in the absence and presence of NOX5 (melittin) and Src (PP2) inhibitors. NOX5 and p22phox-dependent NOXs (NOX1–4) were down-regulated using NOX5 siRNA and p22phox-siRNA approaches. As proof of concept in intact vessels, vascular function was assessed by myography in transgenic mice expressing human NOX5 in a VSMC-specific manner. In HT VSMCs, NOX5 was up-regulated, with associated oxidative stress, hyperoxidation (c-Src, peroxiredoxin, DJ-1), and hyperphosphorylation (c-Src, PKC, ERK1/2, MLC20) of signalling molecules. NOX5 siRNA reduced ROS generation in NT and HT subjects. NOX5 siRNA, but not p22phox-siRNA, blunted c-Src phosphorylation in HT VSMCs. NOX5 siRNA reduced phosphorylation of MLC20 and FAK in NT and HT. In p22phox- silenced HT VSMCs, Ang II-induced phosphorylation of MLC20 was increased, effects blocked by melittin and PP2. NOX5 and c-Src inhibition attenuated actin polymerization and migration in HT VSMCs. In NOX5 transgenic mice, vascular hypercontractilty was decreased by melittin and PP2. We define NOX5/ROS/c-Src as a novel feedforward signalling network in human VSMCs. Amplification of this system in hypertension contributes to VSMC dysfunction. Dampening the NOX5/ROS/c-Src pathway may ameliorate hypertension-associated vascular injury.
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DOI：
10.1172/jci97490
发表时间：
2018-07-02
期刊：
The Journal of clinical investigation
影响因子：
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Emmerson A;Trevelin SC;Mongue-Din H;Becker PD;Ortiz C;Smyth LA;Peng Q;Elgueta R;Sawyer G;Ivetic A;Lechler RI;Lombardi G;Shah AM
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