Abstract
Renin–angiotensin system inhibitors significantly reduce the incidence of arrhythmias. However, the underlying mechanism(s) is not well understood. We aim to test the hypothesis that angiotensin II (Ang II) induces early afterdepolarizations (EADs) and triggered activities (TAs) via the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase–ROS–calmodulin kinase II (CaMKII) pathway. ROS production was analyzed in the isolated rabbit myocytes loaded with ROS dye. Ang II (1–2μM) increased ROS fluorescence in myocytes, which was abolished by Ang II type 1 receptor blocker losartan, NADPH oxidase inhibitor apocynin, and antioxidant MnTMPyP, respectively. Action potentials were recorded using the perforated patch-clamp technique. EADs emerged in 27 out of 41 (66%) cells at 15.8±1.6min after Ang II (1–2μM) perfusion. Ang II-induced EADs were eliminated by losartan, apocynin, or trolox. The CaMKII inhibitor KN-93 (n=6) and inhibitory peptide (AIP) (n=4) also suppressed Ang II-induced EADs, whereas the inactive analogue KN-92 did not. Nifedipine, a blocker of L-type Ca current (ICa2+,L), or ranolazine, an inhibitor of late Na current (INa+), abolished Ang II-induced EADs. The effects of Ang II on major membrane currents were evaluated using voltage clamp. While Ang II at same concentrations had no significant effect on total outward K+ current, it enhanced ICa.L and late INa, which were attenuated by losartan, apocynin, trolox, or KN-93. We conclude that Ang II induces EADs via intracellular ROS production through NADPH oxidase, activation of CaMKII, and enhancement of ICa,L and late INa. These results provide evidence supporting a link between renin–angiotensin system and cardiac arrhythmias.
► Ang II induces early afterdepolarizations (EADs) and triggered activities in rabbit myocytes. ► The NADPH oxidase-ROS-calmodulin kinase II (CaMKII) pathway is involved. ► The Ang II-induced EADs are caused by the activation of both ICa,L and late INa mediated by CaMKII. ► These results support a link between renin-angiotensin system and cardiac arrhythmias.