Salvianolic acid B and Senkyunolide I synergistically alleviate cardiac hypertrophy and regulate MAP3K1 signaling.

Background: Cardiac hypertrophy, characterized by the thickening of the heart muscle, arises from factors such as hypertension and genetic mutations, often leading to adverse outcomes like heart failure and arrhythmias. Guanxinning tablets (GXNT), a botanical drug composed of the blood-activating herbs Salvia miltiorrhiza Bunge. and Ligusticum striatum DC., are widely used in the treatment of cardiovascular diseases. However, the active ingredients and their molecular mechanisms are yet to be fully understood.

Methods: We evaluated the anti-hypertrophic effects of GXNT and screened its active substances via cardiac function live-imaging on an aristolochic acid A-stimulated zebrafish cardiac hypertrophy model, and through F-actin immunostaining on a phenylephrine-induced hypertrophic NRCMs model. Additionally, the protective effects of GXNT's active substances were analyzed in a mouse model of cardiac hypertrophy using echocardiography, histopathology analysis, and western blotting.

Results: The anti-hypertrophic effects of GXNT were assessed using an aristolochic acid A-stimulated zebrafish model and phenylephrine-induced hypertrophic NRCMs. GXNT demonstrated significant anti-hypertrophic effects in both models. Phenotypic screening identified Senkyunolide I (Sen I) from Ligusticum striatum as the active component in the zebrafish model, while Salvianolic acid B (Sal B) and Rosmarinic acid from Salvia miltiorrhiza emerged as the key anti-hypertrophic compound in NRCMs. In a mouse model of isoproterenol-induced cardiac hypertrophy, Sal B and Sen I showed synergistic effects, improving cardiac function, reducing oxidative stress, and suppressing inflammation. Mechanistically, transcriptomic sequencing highlighted cooperative modulation of MAP3K1 signaling by the two compounds. Notably, siRNA-mediated knockdown of MAP3K1 in cardiomyocytes attenuated the hypertrophic phenotype, supporting its essential role in the pathological process. Molecular docking and dynamic simulations further supported their binding potential to MAP3K1.

Conclusion: These findings underscore GXNT's potent anti-hypertrophic effects, possibly driven by the synergistic actions of Sal B and Sen I, and offer insights into its therapeutic potential through MAP3K1 signaling regulation.