Fisetin Attenuates Doxorubicin-Induced Cardiomyopathy In Vivo and In Vitro by Inhibiting Ferroptosis Through SIRT1/Nrf2 Signaling Pathway Activation

Danlei Li,Xiaoman Liu,Lei Yu,Cheng Xu,Zhenzhu Sun,Jianjun Jiang,Wenhu Pi,Yang Zhang

doi:10.3389/fphar.2021.808480

Danlei Li, Xiaoman Liu + Show 6 more

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https://doi.org/10.3389/fphar.2021.808480

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Abstract

Doxorubicin (DOX) is an anthracycline antibiotic that is used extensively for the management of carcinoma; however, its clinical application is limited due to its serious cardiotoxic side effects. Ferroptosis represents iron-dependent and reactive oxygen species (ROS)-related cell death and has been proven to contribute to the progression of DOX-induced cardiomyopathy. Fisetin is a natural flavonoid that is abundantly present in fruits and vegetables. It has been reported to exert cardioprotective effects against DOX-induced cardiotoxicity in experimental rats. However, the underlying mechanisms remain unknown. The present study investigated the cardioprotective role of fisetin and the underlying molecular mechanism through experiments in the DOX-induced cardiomyopathy rat and H9c2 cell models. The results revealed that fisetin treatment could markedly abate DOX-induced cardiotoxicity by alleviating cardiac dysfunction, ameliorating myocardial fibrosis, mitigating cardiac hypertrophy in rats, and attenuating ferroptosis of cardiomyocytes by reversing the decline in the GPX4 level. Mechanistically, fisetin exerted its antioxidant effect by reducing the MDA and lipid ROS levels and increasing the glutathione (GSH) level. Moreover, fisetin exerted its protective effect by increasing the SIRT1 expression and the Nrf2 mRNA and protein levels and its nuclear translocation, which resulted in the activation of its downstream genes such as HO-1 and FTH1. Selective inhibition of SIRT1 attenuated the protective effects of fisetin in the H9c2 cells, which in turn decreased the GSH and GPX4 levels, as well as Nrf2, HO-1, and FTH1 expressions. In conclusion, fisetin exerts its therapeutic effects against DOX-induced cardiomyopathy by inhibiting ferroptosis via SIRT1/Nrf2 signaling pathway activation.

Highlights

Doxorubicin (DOX), a highly effective antitumor anthracycline antibiotic derived from Streptomyces, is commonly used for the treatment of many types of malignancies, including solid tumors, breast cancer, soft tissue sarcomas, lymphomas, and leukemia (Carvalho et al, 2009)
DOX caused cardiac hypertrophy, which was determined through wheat germ agglutinin (WGA) staining
We found that the Nrf2 protein level decreased (Supplementary Figure S3A); the SIRT1 expression level was not significantly decreased whereas Glutathione peroxidase 4 (GPX4), heme oxygenese-1 (HO-1), and ferritin heavy chain 1 (FTH1) expression levels decreased in the Nrf2-knockdown cells treated with DOX + fisetin (Supplementary Figures S3C–D)

Summary

Introduction

Doxorubicin (DOX), a highly effective antitumor anthracycline antibiotic derived from Streptomyces, is commonly used for the treatment of many types of malignancies, including solid tumors, breast cancer, soft tissue sarcomas, lymphomas, and leukemia (Carvalho et al, 2009). The underlying mechanisms of DOX-induced cardiotoxicity are thought to be complex and multifactorial that involve increased lipid peroxidation, oxidative stress, DNA/RNA damage, autophagy and apoptosis inhibition, and calcium homeostasis disturbance (Octavia et al, 2012; Li et al, 2016; Abdel-Daim et al, 2017). Growing evidence suggests that the overproduction of reactive oxygen species (ROS) and increased oxidant-induced mitochondrial damage are crucial for the development of DOX-induced cardiotoxic effects (Christiansen 2011; Carvalho et al, 2014). ROS and mitochondrial dysfunction are the key factors for ferroptosis, which is a newly discovered type of regulated cell death characterized by the accumulation of a large amount of iron and lipid peroxidation in cells (Dixon et al, 2012). Researchers have discovered that natural flavonoids could be used to ameliorate the DOX-induced cardiotoxicity based on flavonoids’ antioxidant, anti-inflammatory, and anti-apoptosis potentials (Guo et al, 2020; Navarro-Hortal et al, 2020; Qi et al, 2020; Sun et al, 2020); supporting evidence for the involvement of flavonoids in DOX-induced ferroptotic myocardial cell death is lacking

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