Mitochondrial mediated thimerosal-induced apoptosis in a human neuroblastoma cell line (SK-N-SH)

Department of Pharmacology, Joan C. Edwards School of Medicine, Marshall University, Huntington, WV 25704-9388, USA.
NeuroToxicology (Impact Factor: 3.05). 07/2005; 26(3):407-16. DOI: 10.1016/j.neuro.2005.03.008
Source: PubMed

ABSTRACT Environmental exposure to mercurials continues to be a public health issue due to their deleterious effects on immune, renal and neurological function. Recently the safety of thimerosal, an ethyl mercury-containing preservative used in vaccines, has been questioned due to exposure of infants during immunization. Mercurials have been reported to cause apoptosis in cultured neurons; however, the signaling pathways resulting in cell death have not been well characterized. Therefore, the objective of this study was to identify the mode of cell death in an in vitro model of thimerosal-induced neurotoxicity, and more specifically, to elucidate signaling pathways which might serve as pharmacological targets. Within 2 h of thimerosal exposure (5 microM) to the human neuroblastoma cell line, SK-N-SH, morphological changes, including membrane alterations and cell shrinkage, were observed. Cell viability, assessed by measurement of lactate dehydrogenase (LDH) activity in the medium, as well as the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, showed a time- and concentration-dependent decrease in cell survival upon thimerosal exposure. In cells treated for 24 h with thimerosal, fluorescence microscopy indicated cells undergoing both apoptosis and oncosis/necrosis. To identify the apoptotic pathway associated with thimerosal-mediated cell death, we first evaluated the mitochondrial cascade, as both inorganic and organic mercurials have been reported to accumulate in the organelle. Cytochrome c was shown to leak from the mitochondria, followed by caspase 9 cleavage within 8 h of treatment. In addition, poly(ADP-ribose) polymerase (PARP) was cleaved to form a 85 kDa fragment following maximal caspase 3 activation at 24 h. Taken together these findings suggest deleterious effects on the cytoarchitecture by thimerosal and initiation of mitochondrial-mediated apoptosis.

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    • "This indicates doserelated mitochondrial potential damage and concurrent apoptosis , as was seen in the previous section. The property of etHg contained in thimerosal disturbing mitochondrial function has been examined mostly in neuronal and immune cell lines (Makani et al., 2002; Humphrey et al., 2005; Yel et al., 2005; Migdal et al., 2010; Zieminska et al., 2010). We believe ours is the first report on the effects of thimerosal on mitochondrial permeability transition in the kidney. "
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    ABSTRACT: Thimerosal is an ethyl mercury-containing compound used mainly in vaccines as a bactericide. Although the kidney is a key target for mercury toxicity, thimerosal nephrotoxicity has not received the same attention as other mercury species. The aim of this study was to determine the potential cytotoxic mechanisms of thimerosal on human kidney cells. Human kidney proximal tubular epithelial (HK2) cells were exposed for 24 h to thimerosal (0-2 µM), and assessed for cell viability, apoptosis, and cell proliferation; expression of proteins Bax, nuclear factor-κB subunits, and transforming growth factor-β1 (TGFβ1); mitochondrial health (JC-1, MitoTracker Red CMXRos); and fibronectin levels (enzyme-linked immunosorbent assay). Thimerosal diminished HK2 cell viability and mitosis, promoted apoptosis, impaired the mitochondrial permeability transition, enhanced Bax and TGFβ1 expression, and augmented fibronectin secretion. This is the first report about kidney cell death and pro-fibrotic mechanisms promoted by thimerosal. Collectively, these in vitro results demonstrate that (1) thimerosal induces kidney epithelial cell apoptosis via upregulating Bax and the mitochondrial apoptotic pathway, and (2) thimerosal is a potential pro-fibrotic agent in human kidney cells. We suggest that new evidence on toxicity as well as continuous surveillance in terms of fibrogenesis is required concerning thimerosal use. © 2014 Wiley Periodicals, Inc. Environ Toxicol, 2014.
    Environmental Toxicology 06/2014; DOI:10.1002/tox.22012 · 3.23 Impact Factor
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    • "The mitochondrion is a primary target of mercury-induced injury and the mitochondrial electron transport chain is the most likely site where excess ROS are generated to induce oxidative stress in mercury toxicity (Yee and Choi, 1996). Mitochondrial effects of mercury in vivo and in vitro, include mitochondrial dysfunction (Chávez and Holguín, 1988; Chávez et al., 1989; Hare and Atchison, 1992; Dreiem et al., 2005; Hernández-Esquivel et al., 2011), membrane permeability transition pore opening (Limke and Atchison, 2002; Polunas et al., 2011) and apoptosis (Shenker et al., 1999; Kim and Sharma, 2004; Humphrey et al., 2005). "
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    ABSTRACT: Occupational or environmental exposures to heavy metals produce several adverse health effects. The common mechanism determining their toxicity and carcinogenicity is the generation of oxidative stress that leads to hepatic damage. In addition, oxidative stress induced by metal exposure leads to the activation of the nuclear factor (erythroid-derived 2)-like 2/Kelch-like ECH-associated protein 1/antioxidant response elements (Nrf2/Keap1/ARE) pathway. Since antioxidant and chelating agents are generally used for the treatment of heavy metals poisoning, this review is focused on the protective role of curcumin against heavy metals liver injury. Curcumin has shown, in clinical and preclinical studies, numerous biological activities including therapeutic efficacy against various human diseases and anti-hepatotoxic effects against environmental or occupational toxins. Curcumin reduces the hepatotoxicity induced by arsenic, cadmium, chromium, copper, lead and mercury, prevents histological injury, lipid peroxidation and glutathione (GSH) depletion, maintains the liver antioxidant enzyme status and protects against mitochondrial dysfunction. The preventive effect of curcumin on the noxious effects induced by heavy metals has been attributed to its scavenging and chelating properties, and/or to the ability to induce the Nrf2/Keap1/ARE pathway. However, additional research is needed in order to propose curcumin as a potential protective agent against liver damage induced by heavy metals.
    Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association 04/2014; 69. DOI:10.1016/j.fct.2014.04.016 · 2.61 Impact Factor
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    • " . 2003 ; Humphrey et al . 2005 ; Kuo et al . 2009 ; Li et al . 2012 ) . Thimerosal was shown to induce DNA and mem - brane damage and also caspase - 3 - dependent apoptosis in the human neuronal and fibro - blasts ( Baskin , Ngo , and Didenko 2003 ) , to activate caspase - 9 and caspase - 3 in T cells and human neuroblasts ( Makani et al . 2002 ; Humphrey et al . 2005 ) , and to activate the c - Jun N - terminal kinase pathway in neuroblastoma cells ( Herdman et al . 2006 ) . Addition - ally , there is a report of thimerosal suppressing the PI 3 K / Akt / survivin pathway which is responsible for apoptosis inhibition in mouse myoblasts ( Li et al . 2012 ) . Similarly , Yel et al . ( 2005 ) reported t"
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    ABSTRACT: Mercury (Hg) is a hazardous chemical that accumulates in many cells and tissues, thereby producing toxicity. The kidney is a key target organ for Hg accumulation and toxicity. The contributing factors to Hg accumulation in humans include: (1) elemental and inorganic Hg exposure, often occurring by inhalation of Hg vapors; (2) exposure to methyl Hg (meHg), for example, through contaminated seafood; and (3) exposure to ethyl mercury (etHg) via thimerosal-containing vaccines. Systematic investigations on the toxic effects of etHg/thimerosal on the nervous system were carried out, and etHg/thimerosal emerged as a possible risk factor for autism and other neurodevelopmental disorders. There is, however, little known about the mechanisms and molecular interactions underlying toxicity of etHg/thimerosal in the kidney, which is the focus of the current review. Susceptible populations such as infants, pregnant women, and the elderly are exposed to etHg through thimerosal-containing vaccines, and in-depth study of the potential adverse effects on the kidney is needed. In general, toxicity occurring in association with different forms of Hg is related to: intracellular thiol metabolism and oxidative stress reactions; mitochondrial function; intracellular distribution and build-up of calcium; apoptosis; expression of stress proteins; and also interaction with the cytoskeleton. Available evidence for the etHg-induced toxicity in the kidney was examined, and the main mechanisms and molecular interactions of cytotoxicity of etHg/thimerosal exposure in kidney described. Such accumulating knowledge may help to indicate molecular pathways that, if modulated, may better handle Hg-mediated toxicity.
    Toxicological and Environmental Chemistry 09/2013; 95(8):1424. DOI:10.1080/02772248.2013.877246 · 0.72 Impact Factor
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