Vascular Nox (NADPH Oxidase) Compartmentalization, Protein Hyperoxidation, and Endoplasmic Reticulum Stress Response in Hypertension.
Vascular Nox (NADPH Oxidase) Compartmentalization, Protein Hyperoxidation, and Endoplasmic Reticulum Stress Response in Hypertension.
复制标题
DOI：
10.1161/hypertensionaha.118.10824
复制
发表时间：
2018-07
期刊：
影响因子：
--
通讯作者：
Touyz RM
中科院分区：
文献类型：
--
作者：
Camargo LL;Harvey AP;Rios FJ;Tsiropoulou S;Da Silva RNO;Cao Z;Graham D;McMaster C;Burchmore RJ;Hartley RC;Bulleid N;Montezano AC;Touyz RM
血管内Nox源性ROS和ER应激与高血压有关。然而，这些过程之间的关系尚不清楚。我们假设，Nox亚型定位在亚细胞区室特异性的方式，有助于氧化和ER应激，影响氧化蛋白质组和血管功能的高血压。在SHR VSMCs中研究了Nox区室化（细胞分级）、O2-（光泽精）、H2O2（Amplex红）、可逆性蛋白氧化（亚磺酰化）、不可逆性蛋白氧化（蛋白酪氨酸磷酸酶（PTP）、过氧化物氧还蛋白氧化）和ER应激（PERK、IRE1 α、磷酸化/氧化）。流式细胞术检测血管平滑肌细胞增殖，肌造影检测血管反应性。siRNA和siRNA均下调Nox表达。在SHR中，Nox定位于特定的亚细胞区域：Nox 1位于质膜，Nox 4位于ER。在SHR中，氧化应激与蛋白质磺酰化增加以及PTP和过氧化物酶的过度氧化有关。抑制Nox 1（NoxA1ds）、Nox 1/4（GKT 137831）和ER应激（4-PBA/Tudca），使SHR血管ROS生成正常化。GKT 137831减少SHR中IRE 1 α亚磺酰化和XBP1剪接。SHR中增加的VSMC增殖通过GKT 137831，4-PBA和STF 083010（IRE1-XBP1破坏剂）正常化。4-PBA可抑制SHRSP的高收缩性。我们证明，高血压中的蛋白质过氧化与氧化和ER应激通过上调质膜-Nox 1和ER-Nox 4。ER应激反应的IRE1-XBP1通路受Nox4/ROS调节，并在SHR VSMC过度增殖表型中起作用。我们的研究强调了Nox亚细胞区室化的重要性以及细胞质ROS和ER应激反应之间的相互作用，这有助于VSMC氧化蛋白质组和高血压血管功能障碍。
Vascular Nox-derived ROS and ER stress have been implicated in hypertension. However relationships between these processes is unclear. We hypothesized that Nox isoforms localize in a sub-cellular compartment-specific manner, contributing to oxidative and ER stress, which influence the oxidative proteome and vascular function in hypertension. Nox compartmentalization (cell fractionation), O2- (lucigenin), H2O2 (amplex red), reversible protein oxidation (sulfenylation), irreversible protein oxidation (protein tyrosine phosphatase (PTP), peroxiredoxin oxidation) and ER stress (PERK, IRE1α, phosphorylation/oxidation) were studied in SHR VSMCs. VSMC proliferation was measured by FACS and vascular reactivity assessed in SHRSP arteries by myography. Noxs were downregulated by siRNA and pharmacologically. In SHR, Noxs were localized in specific sub-cellular regions: Nox1 in plasma-membrane and Nox4 in ER. In SHR, oxidative stress was associated with increased protein sulfenylation and hyperoxidation of PTPs and peroxiredoxins. Inhibition of Nox1 (NoxA1ds), Nox1/4 (GKT 137831) and ER stress (4-PBA/Tudca), normalized SHR vascular ROS generation. GKT137831 reduced IRE1α sulfenylation and XBP1 splicing in SHR. Increased VSMC proliferation in SHR was normalized by GKT137831,4-PBA and STF083010, (IRE1-XBP1 disruptor). Hypercontractility in SHRSP was attenuated by 4-PBA. We demonstrate that protein hyperoxidation in hypertension is associated with oxidative and ER stress through upregulation of plasmalemmal-Nox1 and ER-Nox4. The IRE1-XBP1 pathway of the ER stress response is regulated by Nox4/ROS and plays a role in the hyperproliferative VSMC phenotype in SHR. Our study highlights the importance of Nox sub-cellular compartmentalization and interplay between cytoplasmic ROS and ER stress response, which contribute to the VSMC oxidative proteome and vascular dysfunction in hypertension.