Abstract
Background: Reactive oxygen species (ROS) and calcium ions (Ca2+) are among the major effectors of Ang II (angiotensin II) in vascular smooth muscle cells. ROS are related to Ca2+signaling or contraction induced by Ang II, but little is known about their detailed functions. Here, NOX (NADPH oxidase), a major ROS source responsive to Ang II, was investigated regarding its contribution to Ca2+signaling. Methods: Vascular smooth muscle cells were primary cultured from rat aorta. Ca2+and ROS were monitored mainly using fura-2 and HyPer family probes‚ respectively. Signals activating NOX were examined with relevant pharmacological inhibitors and genetic manipulation techniques. Results: Ang II-induced ROS generation was found to be biphasic: the first phase of ROS production, which was mainly mediated by NOX1, was small and transient, preceding a rise in Ca2+, and the second phase of ROS generation, mediated by NOX1 and NOX4, was slow but sizeable, continuing over tens of minutes. NOX1-derived superoxide in the first phase is required for Ca2+influx through nonselective cation channels. AT1R (Ang II type 1 receptor)-Gβγ-PI3Kγ(phosphoinositide 3-kinase γ) signaling pathway was responsible for the rapid activation of NOX1 in the first phase, while in the second phase, NOX1 was further activated by a separate AT1R-Gαq/11-PLC (phospholipase C)-PKCβ(protein kinase C β) signaling axis. Consistent with these observations, aortas from NOX1-knockout mice exhibited reduced contractility in response to Ang II, and thus the acute pressor response to Ang II was also attenuated in NOX1-knockout mice. Conclusions: NOX1 mediates Ca2+signal generation and thereby contributes to vascular contraction and blood pressure elevation by Ang II.
Original language | English |
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Pages (from-to) | E117-E130 |
Journal | Arteriosclerosis, Thrombosis, and Vascular Biology |
Volume | 42 |
Issue number | 5 |
DOIs | |
State | Published - 1 May 2022 |
Keywords
- angiotensin II
- calcium
- NADPH oxidases
- reactive oxygen species
- smooth muscle