Huashi Baidu formula may attenuate adriamycin-induced myocardial injury via modulating NRF2-ATF3-SRXN1 axis-mediated ferroptosis

Abstract

Objective

Myocardial injury is a fatal adverse effect of adriamycin (ADR), which greatly limits its clinical application. Numerous studies have shown that Huashi Baidu formula (HBF) may exert a satisfactory effect on myocardial function repair. This study aims to reveal the underlying mechanisms and main bioactive compounds (BACs) of HBF against myocardial injury.

Methods

Firstly, this study established an ADR-induced mouse model to assess the protective effects of HBF against myocardial injury. Subsequently, transcriptomic profiling was conducted to screen the differentially expressed genes among the HBF treatment, ADR-induced model, and normal control groups. Following the construction and analysis of protein–protein interactions, gene set enrichment analysis, and co-expression matrix, the key targets of HBF against myocardial injury were screened and further validated based on the ADR-induced mouse model. An integrative approach combining network mapping, molecular docking, and molecular dynamics simulation was performed to identify the representative BACs.

Results

Based on an ADR-induced myocardial injury mouse model, HBF significantly ameliorated myocardial injury by improving body weight loss, decreasing mortality, and rescuing severe cardiac fibrosis. A total of 361 ADR-related genes and 42 HBF therapeutic effect-related genes were identified with the thresholds P < 0.05 and fold change (FC) > 1.2 or < 0.833. Transcriptomic profiling-based networks demonstrated that the nuclear factor erythroid 2-related factor 2 (NRF2)-cyclic AMP-dependent transcription factor ATF-3 (ATF3)-sulfiredoxin-1 (SRXN1) axis-mediated ferroptosis was the key target of HBF against myocardial injury. In vivo validation showed that HBF treatment effectively inhibited serum enzymatic biomarkers of general myocardial injury (aspartate aminotransferase, lactate dehydrogenase, and creatine kinase), myofibroblast activity (transforming growth factor beta 1 and α-smooth muscle actin), inflammatory cytokines (tumor necrosis factor-α and interleukin-6), oxidative damage (superoxide dismutase, malondialdehyde, accumulated iron content, and TUNEL-positive cells). Mechanically, HBF may restore the dysregulation of the NRF2-ATF3-SRXN1 signal axis, leading to inhibition of ferroptosis-associated protein expression. Emodin, rhein, and lactiflorin were identified as the underlying representative BACs of HBF against myocardial injury due to the strong binding affinities between candidate BACs and key targets [Kelch-like ECH-associated protein 1 (KEAP1), ATF3, SRXN1, system Xc- (SLC3A2/SLC7A11), glutathione peroxidase 4 (GPX4), ferroportin (FPN), glutathione synthetase (GSS)]. Molecular dynamics simulations further verified these findings, revealing the stable binding among the three components and the key targets.

Conclusion

HBF may attenuate ADR-induced myocardial injury via modulating NRF2-ATF3-SRXN1 axis-mediated ferroptosis, and emodin, rhein, and lactiflorin are potential representative BACs.