Elsevier

Toxicology

Volume 212, Issues 2–3, 1 September 2005, Pages 155-164
Toxicology

The protective effect of caffeic acid phenethyl ester against cyclosporine A-induced cardiotoxicity in rats

https://doi.org/10.1016/j.tox.2005.04.020Get rights and content

Abstract

Cyclosporine A (CsA) is the immunosuppressor, which is most frequently used in transplant surgery and in the treatment of autoimmune diseases. Oxidative stress has been considered as one of the possible mechanisms of CsA-induced cardiotoxicity. The present investigation examines the ability of caffeic acid phenethyl ester (CAPE), which is an active component of propolis extracts, as a natural antioxidant to protect against CsA-induced oxidative stress and cardiotoxicity. CsA cardiotoxicity was induced by subcutaneous injection of CsA at a dose of 15 mg/kg/body weight daily for 21 days in rats. Cardiotoxicity was evaluated by morphological and biochemical studies. CsA treated rats showed degenerative changes with cardiac fibrosis localized around the fibers. These latters were disorganised and the network was disappeared. The ROS production was increased whereas cytochrome-c-oxidase decreased. The expression and levels of matrix metalloproteinase 2 (MMP2) were increased whereas those of its inhibitor were downregulated. CAPE subcutaneous administration (15 μmol/kg/day) improved cardiac cytoarchitecture, decreased the levels and the expression of MMP2, and increased those of TIMP2 proteins. Moreover, it increased cytochrome-c-oxidase activity and decreased ROS production. These results suggest that CAPE could have protective effect against CsA-induced cardiotoxicity.

Introduction

The use of cyclosporine A (CsA) as an immunosuppressant and anti-rejection drug has been related to nephrotoxicity (Rezzani et al., 2002), hypertension (Nishiyama et al., 2003), hepatotoxicity (Durak et al., 2004, Rezzani, 2004) and cardiotoxicity (Bianchi et al., 2003, Rezzani et al., 2003). Studies to elucidate the mechanism(s) of these adverse effects have implicated free radical production (Zachariae, 1999), lipid peroxidation (Wang and Salahudeen, 1995), induction of the cytochrome P450 (CYP450) system (Mayer et al., 1989) and increased synthesis of vasoconstrictor eicosanoids, mainly tromboxane A2 (TxA2) (Perico et al., 1986, Bianchi et al., 2003). CsA is also an inducer of the powerful vasoconstrictor endothelin (ET), whose levels are elevated after CsA treatment in experimental and clinical conditions (Rezzani et al., 2001, Cauduro et al., 2004). Although mechanisms by which CsA induced toxicity are still not clear and still matter of debate, several reports have suggested that oxidative stress is a possible pathway of CsA toxicity (Grieve and Shah, 2003, Rezzani et al., 2003). Oxidative stress is defined as a tissue injury referring to an imbalance between the production of reactive oxygen species (ROS) and endogenous antioxidant defence systems. Under these conditions, the endogenous antioxidants such as catalase, superoxide dismutase, glutathione and cytochrome-c-oxidase, may be unable to counter ROS formation. So, their presence may cause cellular damage by peroxidation of membrane lipids, sulphydryl enzyme inactivation, protein crosslinking and DNA breakdown (Ilhan et al., 2004). Moreover, it is known that, in heart failure, the ROS production activates latent resident matrix metalloproteinase (MMP) enzymes (Siwik and Colucci, 2004) and induces a decrease in their endogenous inhibitors (TIMPs). The MMPs are a family of enzymes capable of matrix component degradation and they are critically important in determining the balance between matrix deposition and degradation, which ultimately influences chamber size, shape and function (Grieve and Shah, 2003). So, the inappropriate activation of MMPs and/or an imbalance between their actions and those of their inhibitors, for example due to ROS increase, are implicated in cardiac remodelling (Spinale, 2002). According to these data, in our previous works, we observed, after experimental CsA treatment, an increase in MMP2 activity and myocardial extracellular matrix (ECM) disorganization with the disappearance of normal network among the fibers (Bianchi et al., 2003). Moreover, other authors, evaluating the dysregulation of the myocardial ECM in different cardiomyopathies, demonstrated an evident MMP2 upregulation with an inhibition of endogenous antioxidant systems (Siwik et al., 2001). So, more attention have been paid to the protective effects of natural antioxidants against drug-induced toxicities (i.e. CsA) especially whenever free radical generations are involved. Caffeic acid phenethyl ester (CAPE), a flavonoid-like compound, is one of the major components of honeybee propolis with several biological and pharmacological properties. It has been demonstrated that CAPE has antioxidant (Ramazan Yilmaz et al., 2004), anti-inflammatory (Michaluart et al., 1999), anti-carcinogenic (Chen et al., 2001), antiviral (Fesen et al., 1994) and immunomodulatory activities (Park and Kang, 1999). Moreover, it prevents lipid peroxidation induced by ischaemia-reperfusion injury in renal tissue, spinal cord and brain (Irmak et al., 2001, Irmak et al., 2003). At a concentration of 15 μmol/kg/day CAPE completely blocks the production of ROS in human neutrophilis and the xanthine/xanthine oxidase system (Sud’ina et al., 1993). Moreover, it inhibits inducible nitric oxide synthase by increasing leukocyte apoptosis and decreasing leukocyte concentration (Song et al., 2002).

The hypothesis tested is that CAPE is able to reduce or block CsA cardiotoxicity by its antioxidant properties. We have also examined, by morphological and biochemical studies, whether side CsA effects might be related to: (1) an imbalance between the activities of MMP2 and its inhibitor, TIMP2; (2) a decrease in antioxidant markers such as cytochrome-c-oxidase.

Section snippets

Drugs

CsA (Sandimmun, Sandoz) and CAPE (Sigma–Aldrich, Italy) were both dissolved in olive oil. Their final concentration was of 15 mg/kg for CsA and 15 μmol/kg for CAPE during the experimental treatment. They were subcutaneously injected daily during all treatment. The CsA and CAPE doses have been chosen, respectively, according to Schuurman et al. (1990) and Doganay et al. (2002).

Animals and experimental groups

This study included 64 adult male Wistar rats weighing 200–250 g. The animals were housed in cages and kept on a 12 h

Results

The morphological and biochemical results were similar in control and CAPE alone treated rats, so we decided to consider them without distinction.

Discussion

In this study, we showed, first, that CsA determined: (1) upregulation of MMP2 and downregulation of TIMP2 mediated by ROS production; (2) decrease in cytochrome-c-oxidase expression and increase in ROS production. In particular, the increase in MMP2 and decrease in TIMP2 expression should be a compensatory mechanism against the excessive accumulation of collagen. However, in this and in our previous work (Bianchi et al., 2003) we observed that this alteration in MMP2 and TIMP2 proteins is not

References (39)

  • R. Rezzani et al.

    Changes in Hsp90 expression determine the effects of cyclosporine A on the NO pathway in rat myocardium

    FEBS Lett.

    (2003)
  • R. Rezzani

    Cyclosporine A and adverse effects on organs: histochemical studies

    Prog. Histoc. Cytoc.

    (2004)
  • Y.S. Song et al.

    Caffeic acid phenethyl ester inhibits nitric oxide synthase gene expression and enzyme activity

    Cancer Lett.

    (2002)
  • G.F. Sud’ina et al.

    Caffeic acid ester as a lipoxygenase inhibitor with antioxidant properties

    FEBS

    (1993)
  • C. Wang et al.

    Lipid peroxidation accompanies cyclosporine nephrotoxicity: effects of Vitamin E

    Kidney Int.

    (1995)
  • Z.P. Yang et al.

    Lipid peroxidation and changes in cytochrome c oxidase and xanthine oxidase activity in organophosphorus anticholinesterase induced myopathy

    J. Physiol. Paris

    (1998)
  • X. Zhang et al.

    Differential vulnerability to oxidative stress in rat cardiac myocytes versus fibroblasts

    J. Am. Coll. Cardiol.

    (2001)
  • M.S. Burstone

    New histochemical techniques for the demonstration of tissue oxidase (cytochrome oxidase)

    J. Histochem. Cytochem.

    (1959)
  • Y.J. Chen et al.

    The antioxidant caffeic acid phenethyl ester induces apoptosis associated with selective scavenging of hydrogen peroxide in human leukemic HL-60 cells

    Anticancer Drugs

    (2001)
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