RNF34 ablation promotes cerebrovascular remodeling and hypertension by increasing NADPH-derived ROS generation
RNF34 ablation promotes cerebrovascular remodeling and hypertension by increasing NADPH-derived ROS generation
复制标题
RNF34 消融通过增加 NADPH 衍生的 ROS 生成促进脑血管重塑和高血压
DOI：
10.1016/j.nbd.2021.105396
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发表时间：
2021-05-28
影响因子：
6.1
通讯作者：
Zhang, Beilin
中科院分区：
文献类型：
--
作者：
Fang, Shaokuan;Cheng, Yingying;Zhang, Beilin
脑血管重构是高血压和中风最常见的原因。泛素E3连接酶环指蛋白34(RNF34)被认为与多种神经系统疾病的发生发展有关。然而，RNF34在脑血管重构和高血压中的重要性却知之甚少。在这里，我们使用了RNF34基因全局敲除的小鼠以及RNF34基因敲除的小鼠来删除内皮细胞和平滑肌细胞(SMC)中的RNF34。我们的结果显示，全球RNF34基因敲除小鼠显著促进血管紧张素II(AngII)诱导的大脑中动脉(MCA)重构、高血压和神经功能障碍。内皮细胞RNF34对高血压的发生发展没有调节作用。相反，SMC RNF34的表达是高血压和大脑中动脉重塑的关键调节因素。RNF34缺失可促进血管紧张素Ⅱ诱导的小鼠脑血管平滑肌细胞(MBVSMCs)的增殖、迁移和侵袭。此外，SMC RNF34缺陷小鼠的MCA和MBVSMCs表现出增加超氧阴离子和活性氧物种(ROS)的产生以及烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶的活性，但对线粒体来源的ROS没有明显影响。RNF34基因敲除促进了p22Phox的表达，导致p22Phox/p47Phox和p22Phox/NOX2结合增加，最终形成NADPH氧化酶复合体。免疫沉淀实验证实RNF34与p22Phox相互作用。RNF34缺失通过抑制泛素介导的降解增加了p22Phox蛋白的稳定性。阻断NADPH氧化酶活性或敲除p22Phox可显著消除RNF34缺失对脑血管重构和高血压的影响。总之，我们的研究表明，SMC RNF34缺乏通过减少p22Phox泛在独立的降解，增加NADPH来源的ROS的产生，促进脑血管SMC的增殖和重塑。
Cerebrovascular remodeling is the most common cause of hypertension and stroke. Ubiquitin E3 ligase RING finger protein 34 (RNF34) is suggested to be associated with the development of multiple neurological diseases. However, the importance of RNF34 in cerebrovascular remodeling and hypertension is poorly understood. Herein, we used mice with a global RNF34 knockout as well as RNF34 floxed mice to delete RNF34 in endothelial cells and smooth muscle cells (SMCs). Our results showed that global RNF34 knockout mice substantially promoted angiotensin II (AngII)-induced middle cerebral artery (MCA) remodeling, hypertension, and neurological dysfunction. Endothelial cell RNF34 did not regulate the development of hypertension. Rather, SMC RNF34 expression is a critical regulator of hypertension and MCA remodeling. Loss of RNF34 enhanced AngII-induced mouse brain vascular SMCs (MBVSMCs) proliferation, migration and invasion. Furthermore, MCA and MBVSMCs from SMC RNF34-deficient mice showed increased superoxide anion and reactive oxygen species (ROS) generation as well as nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity, but exhibited no marked effect on mitochondria-derived ROS. Knockout of RNF34 promoted p22phox expression, leading to increased binding of p22phox/p47phox and p22phox/NOX2, and eventually NADPH oxidase complex formation. Immunoprecipitation assay identified that RNF34 interacted with p22phox. RNF34 deletion increased p22phox protein stability by inhibiting ubiquitin-mediated degradation. Blockade of NADPH oxidase activity or knockdown of p22phox significantly abolished the effects of RNF34 deletion on cerebrovascular remodeling and hypertension. Collectively, our study demonstrates that SMC RNF34 deficiency promotes cerebrovascular SMC hyperplasia and remodeling by increased NADPH-derived ROS generation via reducing p22phox ubiquitindependent degradation.