ClC-3 promotes angiotensin II-induced reactive oxygen species production in endothelial cells by facilitating Nox2 NADPH oxidase complex formation

Abstract

Recent evidence suggests that ClC-3, a member of the ClC family of Cl channels or Cl/H antiporters, plays a critical role in NADPH oxidase-derived reactive oxygen species (ROS) generation. However, the underling mechanisms remain unclear. In this study we investigated the effects and mechanisms of ClC-3 on NADPH oxidase activation and ROS generation in endothelial cells. Treatment with angiotensin II (Ang II, 1 μmol/L) significantly elevated ClC-3 expression in cultured human umbilical vein endothelial cells (HUVECs). Furthermore, Ang II treatment increased ROS production and NADPH oxidase activity, an effect that could be significantly inhibited by knockdown of ClC-3, and further enhanced by overexpression of ClC-3. SA-β-galactosidase staining showed that ClC-3 silencing abolished Ang II-induced HUVEC senescence, whereas ClC-3 overexpression caused the opposite effects. We further showed that Ang II treatment increased the translocation of p47phox and p67phox from the cytosol to membrane, accompanied by elevated Nox2 and p22phox expression, which was significantly attenuated by knockdown of ClC-3 and potentiated by overexpression of ClC-3. Moreover, overexpression of ClC-3 increased Ang II-induced phosphorylation of p47phox and p38 MAPK in HUVECs. Pretreatment with a p38 inhibitor SB203580 abolished ClC-3 overexpression-induced increase in p47phox phosphorylation, as well as NADPH oxidase activity and ROS generation. Our results demonstrate that ClC-3 acts as a positive regulator of Ang II-induced NADPH oxidase activation and ROS production in endothelial cells, possibly via promoting both Nox2/p22phox expression and p38 MAPK-dependent p47phox/p67phox membrane translocation, then increasing Nox2 NADPH oxidase complex formation.

SARS-CoV-2 Mediated Endothelial Dysfunction: The Potential Role of Chronic Oxidative Stress

“…Given the extensive damage mtROS can cause, it is critical to examine how SARS-CoV-2 infection can induce mitochondrial dysfunction in ECs. The increase in Ang II levels may be one of the mechanisms responsible for elevating mtROS production and subsequent EC pathologies, as Ang II has been shown to increase ROS production via increased NADPH oxidase (Nox) activation ( Liang et al, 2018 ). Past studies have found that a 4-h, 200 nmol/L treatment of Ang II increased mitochondrial H 2 O 2 production by 1.5-2 times in ECs, an increase mediated through Nox-generated ROS ( Doughan et al, 2008 ).…”

Clcn3 deficiency ameliorates high-fat diet-induced obesity and adipose tissue macrophage inflammation in mice

“…The guinea pig cDNA of Clcn3 was generously gifted by Dr. Dayue Duan (University of Nevada, Reno, USA) for constructing Ad-Clcn3. Adenoviral infection was carried out as previously described [16]. The adenoviral vector (Vector) without expressing Clcn3 was employed as the negative control.…”

Oxidative Stress Links Aging‐Associated Cardiovascular Diseases and Prostatic Diseases

“…The previous studies have reported that the expression of mRNA in NOX1, NOX2, and NOX4 was increased in aortas from animals infused with Ang II [85,86]. In addition, the expression of NOX2, NOX4, and NOX5 was found to be upregulated by Ang II in endothelial cells [87,88], and knockout of NOX1, NOX2, and NOX4 in mice can reduce blood-pressure elevation induced by Ang II [86,89,90]. Taking together, these data suggest that activation of NOX1, NOX2, NOX4, and NOX5 play an important role in the development of Ang II-induced hypertension.…”