A chimeric natriuretic peptide (Ev-NP) inhibits isoproterenol (ISO)-induced hypertrophic growth in in vivo and in vitro models by enhancing cGMP and its downstream signaling targets: In silico docking and binding efficacy analysis with NPR-A and NPR-B receptors

A chimeric natriuretic peptide (Ev-NP) was engineered and created with an intention of having a dual NPR-A/NPR-B activation, resistance to degradation, and with a strong renal, and anti-hypertrophic actions in the heart. In the present study, we aim to investigate the anti-hypertrophic properties of a novel chimeric natriuretic peptide, Ev-NP (37 amino acids), against isoproterenol (ISO)-induced hypertrophy in H9c2 cells in vitro and in a rat model in vivo. The effects on anti-hypertrophy and cGMP stimulation were evaluated in H9c2 cells exposed to ISO, both with and without Ev-NP, at concentrations ranging from 10 to 50 nM over 24 h. A significant dose-dependent increase in cGMP was observed in Ev-NP-treated H9c2 cells compared to controls. Furthermore, Ev-NP treatment significantly (P < 0.001) decreased ISO-induced hypertrophic growth in H9c2 cells by elevating cGMP levels. In H9c2 cells overexpressing Npr1 and co-treated with Ev-NP, a stronger anti-hypertrophic effect was observed, as demonstrated by a significant reduction (P < 0.001) in hypertrophic marker gene expression (α-sk, BNP, and β-MHC) compared to cells treated only with ISO. Furthermore, cytokine array analysis showed that Ev-NP treatment normalized ISO-induced up-regulation of pro-inflammatory and growth factor proteins in H9C2 cells. The in vivo anti-hypertrophic study also showed that Ev-NP significantly reduced (90%) the hypertrophic growth caused by ISO in Wistar rats. Importantly, treatment with Ev-NP restored the ISO-induced reductions in cGMP and NPR-A levels in the rat hearts. In silico analysis revealed that Ev-NP exhibited a stronger affinity for the NPR-A receptor, with a binding energy of -490.17 kcal/mol, compared to NPR-B binding energy of -390.77 kcal/mol. The native ANP exhibited a binding energy of -314.68 kcal/mol with NPR-A. These findings suggest that Ev-NP has promising anti-hypertrophic properties, and its therapeutic potential can be harnessed to treat and manage cardiac hypertrophy and heart failure in humans.