Laboratory of Pharmacology and Toxicology, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, PR China.
Can J Cardiol. 2013 Oct;29(10):1310-9. doi: 10.1016/j.cjca.2013.04.026. Epub 2013 Jul 11.
Activation of epidermal growth factor receptor (EGFR) plays an important role in angiotensin II (Ang II)-induced cardiac hypertrophy, but little is known about the underlying mechanism that results in EGFR activation. In this study, we aimed to confirm the important role of nicotinamide adenine dinucleotide phosphate oxidase 4 (Nox4) in Ang II-induced EGFR activation and subsequent cardiac hypertrophy by upregulating expression of a disintegrin and metalloproteinase (MMP)-17 (ADAM17).
Small interference RNA (siRNA) was adopted to knock down ADAM17 or Nox4 expression. Nox4 plasmid was used to construct cardiomyocytes with Nox4 overexpression.
Nox4 and ADAM17 increased in an abdominal artery coarctation-induced model of myocardial hypertrophy. In vitro studies showed that Nox4 was required in Ang II-induced EGFR activation and subsequent myocardial hypertrophy. Nox4 siRNA and Nox4 overexpression demonstrated that Nox4 controlled the transcription and translation of ADAM17. Furthermore, we observed that the ratio of phosphor-EGFR (p-EGFR) to EGFR was significantly reduced by ADAM17 siRNA in hypertrophic cardiomyocytes. Enzyme-linked immunosorbent assay studies revealed that Nox4 and ADAM17 mediated the release of mature heparin-binding EGF-like growth factor (HB-EGF), which played a critical role in the Ang II-induced EGFR activation. Moreover, the results of reactive oxygen species (ROS) scavenging by N-acetyl cysteine (NAC) indicated that ROS were required in the Nox4-mediated upregulation of ADAM17 expression.
In summary, Nox4 is required in Ang II-induced EGFR activation and subsequent cardiac hypertrophy; it increased the expression of ADAM17, which induced the release of mature HB-EGF. ROS were also required in the Nox4-mediated upregulation of ADAM17 expression.
表皮生长因子受体(EGFR)的激活在血管紧张素 II(Ang II)诱导的心肌肥厚中起着重要作用,但导致 EGFR 激活的潜在机制知之甚少。在这项研究中,我们旨在通过上调金属蛋白酶 17(MMP-17,也称为 ADAM17)的表达,证实烟酰胺腺嘌呤二核苷酸磷酸氧化酶 4(Nox4)在 Ang II 诱导的 EGFR 激活和随后的心肌肥厚中的重要作用。
采用小干扰 RNA(siRNA)敲低 ADAM17 或 Nox4 的表达。使用 Nox4 质粒构建 Nox4 过表达的心肌细胞。
Nox4 和 ADAM17 在腹主动脉缩窄诱导的心肌肥厚模型中增加。体外研究表明,Nox4 在 Ang II 诱导的 EGFR 激活和随后的心肌肥厚中是必需的。Nox4 siRNA 和 Nox4 过表达表明,Nox4 控制 ADAM17 的转录和翻译。此外,我们观察到在肥大的心肌细胞中,ADAM17 siRNA 显著降低了 p-EGFR 与 EGFR 的比值。酶联免疫吸附试验研究表明,Nox4 和 ADAM17 介导了成熟肝素结合表皮生长因子样生长因子(HB-EGF)的释放,这在 Ang II 诱导的 EGFR 激活中起着关键作用。此外,N-乙酰半胱氨酸(NAC)清除活性氧(ROS)的结果表明,ROS 在 Nox4 介导的 ADAM17 表达上调中是必需的。
总之,Nox4 在 Ang II 诱导的 EGFR 激活和随后的心肌肥厚中是必需的;它增加了 ADAM17 的表达,从而诱导了成熟 HB-EGF 的释放。ROS 也在 Nox4 介导的 ADAM17 表达上调中是必需的。