Effects of vericiguat on redox signaling in cardiomyocyte

Abstract Background Reactive oxygen species (ROS) inhibits the nitric oxide (NO)-soluble guanylate cyclase (sGC)-cyclic guanosine monophosphate (cGMP) pathway in the patients with advanced heart failure (HF) and contributes to the prognosis of HF. Vericiguat stimulates sGC independently of NO and reduced morbidity/mortality for HF patients with reduced ejection fraction. However, the effects of vericiguat on the cardiomyocyte in terms of ROS is unclear. Purpose We explored the cardioprotective effects of vericiguat focusing on the oxidative stress. Methods Cardiomyocytes isolated from the whole heart of neonatal rats were incubated with vericiguat (0μM as control, 0.1μM, 1.0μM, 10μM) for 72 hours. Administration of angiotensin II (1.0μM) was used to induce excessive ROS. Superoxide was quantified using luminometry, and whole ROS or mitochondria-derived superoxide was evaluated by using live-cell fluorescent probes. ERK 1/2 signaling pathway activity were evaluated with western blot analysis. Results Nicotinamide adenine dinucleotide phosphate (NADPH)-stimulated superoxide was significantly increased by the treatment of angiotensin II (40208 ± 6107 reactive light unit (RLU) / mg, p = 0.0009) and the angiotensin II-induced superoxide was significantly attenuated by vericiguat at doses of 0.1μM (28240 ± 6507 RLU / mg, p = 0.03) , 1.0μM (25221 ± 6496 RLU / mg, p = 0.007) and 10μM (8230 ± 2568 RLU / mg, p < 0.0001). Corrected total cell fluorescence value of live-cell fluorescent probes revealed that whole ROS and mitochondria-derived superoxide were significantly increased by angiotensin II treatment (525.3 ± 119.6, p = 0.004 and 945.3 ± 153.0, p <0.0001), which were attenuated by vericiguat (0.1μM, 431.2 ± 45.7, p = 0.48 and 548.1 ± 103.9, p = 0.0004; 1.0μM, 336.1 ± 109.0, p = 0.04 and 421.3 ± 111.9, p < 0.0001; 10μM, 270.0 ± 44.4, p = 0.005 and 157.7 ± 48.3, p < 0.0005) dose-dependently. Angiotensin II-induced phosphorylation of ERK 1/2 was suppressed by vericiguat at doses of 1.0μM (fold change 0.58, p = 0.04) and 10μM (fold change 0.45, p = 0.006) treatment. Conclusion Our results indicated that vericiguat could suppress the oxidative stress induced by angiotensin II and following activation of ERK 1/2 signaling. Suppressing the oxidative stress could lead to further activation of NO-sGC-cGMP pathway not only the stimulation by vericiguat and favorable cardio-protective cycle is expected.Quantification of superoxideQuantification of ROS or superoxide