Lujiao Formula attenuates cardiac hypertrophy by modulating AMPK-SIRT1 and PI3K-Akt signaling pathways

Abstract

Cardiac hypertrophy (CH) represents a fundamental morphological adaptation that precedes and drives the transition to heart failure (HF). The traditional formulation Lujiao Formula (LJF) has been widely prescribed in clinical practice for more than two decades to alleviate HF, largely through suppressing CH and ventricular remodeling. Yet, the molecular mechanisms that mediate its cardioprotective benefits remain incompletely understood. In this study, we sought to elucidate both the therapeutic efficacy and mechanistic basis of LJF against CH by integrating metabolomic and proteomic profiling with targeted experimental validation. A murine model of CH was established using transverse aortic constriction (TAC), while complementary cellular models were induced via angiotensin II (Ang II) stimulation. The anti-hypertrophic activity of LJF was evaluated through echocardiography, heart weight index measurements, and histopathological assessment. Expression of canonical hypertrophic markers, including ANP, BNP, and β-MHC, as well as key regulatory proteins, was quantified by qRT-PCR and Western blotting analysis. LJF treatment markedly reduced left ventricular end-diastolic volume (LVEDV), end-systolic volume (LVESV), and systolic internal diameter (LVIDs), while significantly enhancing ejection fraction (EF) and fractional shortening (FS), thereby mitigating maladaptive myocardial remodeling in vivo. Integrative metabolomic and proteomic analyses highlighted AMPK and PI3K–Akt signaling as principal pathways mediating LJF's cardioprotective effects. Mechanistic investigations confirmed that LJF downregulated ANP, BNP, and β-MHC expression, suppressed PI3K and Akt phosphorylation and concurrently upregulated p-AMPKα1/α2 and SIRT1 protein levels both in vivo and in vitro. Importantly, these beneficial effects were partially abrogated by co-treatment with Compound C (CC, an AMPK inhibitor) or LY294002 (a PI3K inhibitor) in vitro. Collectively, our findings demonstrated that LJF exerted significant anti-hypertrophic effects through coordinated modulation of the AMPK–SIRT1 and PI3K–Akt cascades, offering a strong mechanistic basis for its clinical utility in managing CH.