Baicalein alleviates doxorubicin-induced cardiotoxicity via suppression of myocardial oxidative stress and apoptosis in mice

doi:10.1016/j.lfs.2015.11.018

Life Sciences

Volume 144, 1 January 2016, Pages 8-18

https://doi.org/10.1016/j.lfs.2015.11.018 Get rights and content

Abstract

Aims

Doxorubicin is a widely used anthracycline derivative anticancer drug. Unfortunately, the clinical use of doxorubicin has the serious drawback of cardiotoxicity. In this study, we investigated whether baicalein, a bioflavonoid, can prevent doxorubicin-induced cardiotoxicity in vivo and we delineated the possible underlying mechanisms.

Main methods

Male BALB/c mice were treated with either intraperitoneal doxorubicin (15 mg/kg divided into three equal doses for 15 days) and/or oral baicalein (25 and 50 mg/kg for 15 days). Serum markers of cardiac injury, histology of heart, parameters related to myocardial oxidative stress, apoptosis and inflammation were investigated.

Key findings

Treatment with baicalein reduced doxorubicin-induced elevation of serum creatine kinase-MB isoenzyme (CK-MB), lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels and ameliorated the histopathological damage. Baicalein restored the doxorubicin-induced decrease in both enzymatic and non-enzymatic myocardial antioxidants and increased the myocardial expression of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1). Further studies showed that baicalein could inverse the Bax/Bcl-2 ratio, suppress doxorubicin-induced p53, cleaved caspase-3 and PARP expression and prevented doxorubicin-induced DNA damage. Baicalein treatment also interferes with doxorubicin-induced myocardial NF-κB signaling through inhibition of IκBα phosphorylation and nuclear translocation of p65 subunit. Doxorubicin elevated iNOS and nitrites levels were also significantly decreased in baicalein treated mice. However, we did not find any significant change (p > 0.05) in the myocardial TNF-α and IL-6 levels in control and treated animals.

Significance

Our finding suggests that baicalein might be a promising molecule for the prevention of doxorubicin-induced cardiotoxicity.

Graphical abstract

Introduction

Doxorubicin is an effective and widely used anthracycline derivative cytotoxic antibiotic. It is used for the treatment of a variety of neoplasm's including leukemias, lymphomas and solid human malignancies [26]. Despite being a highly targeted molecule for cancer therapy, the clinical use of doxorubicin is limited because of cardiotoxicity leading to dilated cardiomyopathy with congestive heart failure [18]. The mechanism involved in doxorubicin-induced cardiotoxicity is complex and multifactorial and is known to involve, at least in part, oxidative stress and mitochondrial dysfunction, which eventually lead to cardiomyocyte death by apoptosis and/or necrosis [27]. Cardiomyocytes are highly prone to reactive oxygen species (ROS)-induced damage because of lesser amount of antioxidant defense, higher density of mitochondria occupancy and high aerobic metabolism when compared to cells of other tissues [11]. It has also been demonstrated that doxorubicin-induced ROS generation activates NF-κB to elicit subsequent apoptosis in cardiomyocytes [8], [10]. Moreover, generation of ROS promotes lipid peroxidation and apoptotic change due to loss of mitochondrial membrane integrity and p53 activity [31]. Although extensive efforts have been made to prevent doxorubicin-induced cardiotoxicity, there is little harmony and agreement in the best approach.

The health benefits of polyphenolic compounds of plant origin have been widely studied. Baicalein (5, 6, 7-trihydroxy flavone) is a major active constituent in the roots of medicinal herb Scutellaria baicalensis Georgi (Lamiaceae family) [33]. The root of S. baicalensis has been used as folk medicine in China for the treatment of inflammatory diseases, chronic hepatitis, bacterial and viral infections, allergy and thrombotic stroke [14]. Baicalein is also known to exhibit therapeutic effect in endotoxin-induced myocardial dysfunction [20] and ameliorates myocardial ischemic/reperfusion injury in rats [16], [36]. It has also been reported that baicalein attenuates ROS generation and cell death during doxorubicin exposure and exhibits cytoprotective effect against doxorubicin-induced cardiomyocyte injury [4]. Although an in vitro study conducted by Chang et al. [4] addressed the beneficial effects of baicalein on doxorubicin-induced cardiomyocyte injury, there is no in vivo study conducted as yet to understand the mechanism of action and beneficial effect of baicalein on heart in doxorubicin-induced cardiotoxic animals. Hence, present study was designed to elucidate the molecular mechanisms underlying the protective effect of baicalein on heart using doxorubicin-induced cardiotoxicity model in mice.

Section snippets

Chemicals, kits and antibodies

Baicalein (≥ 98.0% purity), doxorubicin, superoxide dismutase (SOD) assay kit, catalase, β-nicotinamide adenine dinucleotide 3-phosphate reduced form (NADPH), 2-thiobarbituric acid (TBA), 1-chloro-2, 4-dinitrobenzene (CDNB), 2, 6-dichlorophenolindophenol (DCIP), reduced glutathione (GSH), 5, 5-dithio-bis (2-nitrobenzoic acid) (DTNB) were purchased from Sigma-Aldrich Co, St. Louis, MO, USA. Halt protease inhibitor cocktail, Bicinchoninic acid (BCA) protein assay kit, NE-PER nuclear and

Effect of baicalein on doxorubicin-induced cardiac injury in mice

As shown in Table 1, in doxorubicin alone treated group of mice, the body weight of mice and the relative weight of heart were significantly (p < 0.001 for body weight; p < 0.05 for relative weight of heart) decreased when compared with the vehicle control mice. A significant (p < 0.001 for CK-MB and AST; p < 0.01 for LDH and ALT) increase in serum levels of CK-MB, LDH, AST and ALT were observed in mice administered with doxorubicin. Interestingly, treatment with baicalein (50 mg/kg) along with

Discussion

The results of the present study demonstrated that the effectiveness of the baicalein in ameliorating doxorubicin-induced cardiotoxicity in mice. Consistent with the changes in serum biochemistry and histological findings caused by doxorubicin in other studies, doxorubicin administration produced a significant elevation of serum CK-MB, LDH, AST and ALT activities and extensive myofibrillar degeneration with severe cytoplasmic vacuolation in heart tissue. The body weight and relative weight of

Conflict of interest statement

The authors declared that there are no conflicts of interest.

Acknowledgments

Authors thank Director, CSIR-IICT for constant support and encouragement. Bidya Dhar Sahu is thankful to Council of Scientific and Industrial Research (CSIR, New Delhi) for providing Senior Research Fellowship.

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Baicalein alleviates doxorubicin-induced cardiotoxicity via suppression of myocardial oxidative stress and apoptosis in mice

Abstract

Aims

Main methods

Key findings

Significance

Graphical abstract

Introduction

Section snippets

Chemicals, kits and antibodies

Effect of baicalein on doxorubicin-induced cardiac injury in mice

Discussion

Conflict of interest statement

Acknowledgments

Biochem. Pharmacol.

Cancer Lett.

Arch. Biochem. Biophys.

Environ. Toxicol. Pharmacol.

Biochem. Pharmacol.

J. Biol. Chem.

Arch. Biochem. Biophys.

J. Ethnopharmacol.

J. Mol. Cell. Cardiol.

Eur. J. Pharmacol.

J. Am. Coll. Cardiol.

Anal. Biochem.

Methods Enzymol.

Biomaterials

Food Chem. Toxicol.

Toxicol. Appl. Pharmacol.

Eur. J. Pharmacol.

J. Biol. Chem.

J. Mol. Cell. Cardiol.

FEBS Lett.

Catalase

Doxorubicin-induced cardiotoxicity: from bioenergetic failure and cell death to cardiomyopathy

Med. Res. Rev.

Baicalein protects against doxorubicin-induced cardiotoxicity by attenuation of mitochondrial oxidant injury and JNK activation

J. Cell. Biochem.