Involvement of ferritin heavy chain in the preventive effect of metformin against doxorubicin-induced cardiotoxicity

Cardiology Department, University Hospital Virgen de la Arrixaca, and Department of Medicine, School of Medicine, University of Murcia, 30120 Murcia, Spain
Free Radical Biology and Medicine (Impact Factor: 5.74). 09/2012; 57. DOI: 10.1016/j.freeradbiomed.2012.09.009.


Doxorubicin is a wide-spectrum chemotherapeutic agent, although a cumulative dose may cause cardiac damage and lead to heart failure. Doxorubicin cardiotoxicity is dependent on the intracellular iron pool and manifests itself by increasing oxidative stress. Our group has recently shown the ability of metformin, an oral antidiabetic with cardiovascular benefits, to protect cardiomyocytes from doxorubicin-induced damage. This work aimed to study whether metformin is able to modulate the expression of ferritin, the major intracellular iron storage protein, in cardiomyocytes and whether it is involved in their protection. The addition of metformin to adult mouse cardiomyocytes (HL-1 cell line) induced both gene and protein expression of the ferritin heavy chain (FHC) in a time-dependent manner. The silencing of FHC expression with siRNAs inhibited the ability of metformin to protect cardiomyocytes from doxorubicin-induced damage, in terms of the percentage of cell viability, the levels of reactive oxygen species, and the activity of antioxidant enzymes (catalase, glutathione peroxidase, and superoxide dismutase). In addition, metformin induced the activation of NF-κB in HL-1 cells, whereas preincubation with SN50, an inhibitor of NF-κB, blocked the upregulation of the FHC and the protective effect mediated by metformin. Taken together, these results provide new knowledge on the protective actions of metformin against doxorubicin-induced cardiotoxicity by identifying FHC and NF-κB as the major mediators of this beneficial effect.

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    • "Determination of reactive oxygen species 24 "
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    ABSTRACT: A key feature of pulmonary hypertension (PH) is the remodeling of small pulmonary arteries due to abnormal pulmonary artery smooth muscle cell (PASMC) proliferation and resistance to apoptosis. However, the cellular mechanisms underlying how PASMCs in the pathological condition of pulmonary hypertension become resistant to apoptosis remain unknown. It was recently reported that lipocalin 2 (Lcn2) is up-regulated in a wide array of malignant conditions, which facilitates tumorigenesis partly by inhibiting cell apoptosis. In this study, we observed that the expression levels of Lcn2 were significantly elevated in a rat PH model induced with monocrotaline and in patients with congenital heart disease-associated PH (CHD-PH) when compared with respective control. Therefore, we hypothesize that Lcn2 could regulate human PASMC (HPASMC) apoptosis through a mechanism. By the detection of DNA fragmentation using the TUNEL assay, the detection of Annexin V/PI-positive cells using flow cytometry, and the detection of cleaved caspase-3 and caspase-3 activity, we observed that Lcn2 significantly inhibited HPASMC apoptosis induced by serum withdrawal and H2O2 treatment. We also observed that Lcn2 down-regulated the proapoptotic protein Bax, decreased the levels of cellular ROS, and up-regulated the expression of superoxide dismutases (SOD1 and SOD2). In conclusion, Lcn2 significantly inhibits HPASMC apoptosis induced by oxidative stress via decreased intracellular ROS and elevated SODs. Up-regulation of Lcn2 in a rat PH model and CHD-PH patients may be involved in the pathological process of PH.
    Preview · Article · Jul 2014 · International journal of biological sciences
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    • "Newly formed ferritin can therefore sequester iron and may paradoxically represent a defense for cardiomyocytes. These findings were recently supported by two studies demonstrating that NF-kB-mediated ferritin H chain induction plays an important role in the mitochondrial protection offered by metformin against DOX cardiotoxicity in isolated cardiomyocytes (Asensio-López et al., 2013a,b). Moreover, it has been shown that exposure of cardiomyocytes to DOX leads to higher accumulation of iron into ferritin by mechanisms that impair iron release from ferritin and/or lysosomal/proteasomal degradation of this iron-storage protein (Kwok and Richardson, 2003, 2004). "
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    ABSTRACT: The clinical use of the antitumor anthracycline Doxorubicin is limited by the risk of severe cardiotoxicity. The mechanisms underlying anthracycline-dependent cardiotoxicity are multiple and remain uncompletely understood, but many observations indicate that interactions with cellular iron metabolism are important. Convincing evidence showing that iron plays a role in Doxorubicin cardiotoxicity is provided by the protecting efficacy of iron chelation in patients and experimental models, and studies showing that iron overload exacerbates the cardiotoxic effects of the drug, but the underlying molecular mechanisms remain to be completely characterized. Since anthracyclines generate reactive oxygen species, increased iron-catalyzed formation of free radicals appears an obvious explanation for the aggravating role of iron in Doxorubicin cardiotoxicity, but antioxidants did not offer protection in clinical settings. Moreover, how the interaction between reactive oxygen species and iron damages heart cells exposed to Doxorubicin is still unclear. This review discusses the pathogenic role of the disruption of iron homeostasis in Doxorubicin-mediated cardiotoxicity in the context of current and future pharmacologic approaches to cardioprotection.
    Full-text · Article · Feb 2014 · Frontiers in Pharmacology
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    ABSTRACT: A substantial minority of type 2 diabetes mellitus (T2DM) patients treated with metformin develop severe gastrointestinal (GI) symptoms leading to drug discontinuation, depriving them of the potentially cardioprotective pleiotropic effects of this first-line oral agent. At present, it is unclear whether treating diabetes without being able to ever use metformin alters cardiovascular outcomes. From a population of 773 consecutive T2DM outpatients, the cardiometabolic phenotypes of 83 patients who discontinued metformin due to GI intolerance (Met-Intol cases) were compared with those of 332 age- and gender-matched metformin-tolerant (Met-Tol) controls, amounting to a case: control ratio of 1:4. Mean age (SD) was 70 (13) (male:female: 46:54). Metformin intolerance was associated with a reduced prevalence of macroangiopathy (P=0.0486), mainly due to a lower prevalence of CAD (-34%; P=0.0374). Met-Intol cases more often belonged to blood group A and subgroup A Rh+, with 50% and 66% relative increases (P=0.0039 and P=0.0005), respectively. There were twice as many non-right-handers among the Met-Intol (18% vs. 9%; P=0.0262), and this group also had significantly higher serum ferritin and LDL cholesterol levels. Statins/fibrates were used by 66%/19% of Met-Tol vs. 48%/18% of Met-Intol (P=0.0051 for statins). On the other hand, there were no differences between groups as regards smoking, diabetes duration, HbA1c, BMI, blood pressure, waist size, fat mass, visceral fat, liver steatosis, the metabolic syndrome, eGFR, albuminuria, erectile dysfunction and microangiopathy. Intolerance to metformin represents an unforeseen phenotype in T2DM patients characterized by a low rate of ischaemic heart disease, left-handedness, ABO group imbalance and an iron load.
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