Melatonin reduces lipid and protein oxidative damage in synaptosomes due to aluminium

doi:10.1016/j.jtemb.2007.04.002

Journal of Trace Elements in Medicine and Biology

Volume 21, Issue 4, 28 November 2007, Pages 261-268

https://doi.org/10.1016/j.jtemb.2007.04.002 Get rights and content

Abstract

Prolonged exposure to excessive aluminium (Al) concentrations is involved in the ethiopathology of certain dementias and neurological disorders. Melatonin is a well-known antioxidant that efficiently reduces lipid peroxidation due to oxidative stress. Herein, we investigated in synaptosomal membranes the effect of melatonin in preventing Al promotion of lipid and protein oxidation when the metal was combined with FeCl₃ and ascorbic acid. Lipid peroxidation was estimated by quantifying malondialdehyde (MDA) and 4-hydroxyalkenals (4-HDA) concentrations in the membrane suspension and protein carbonyls were measured in the synaptosomes as an index of oxidative damage. Under our experimental conditions, the addition of Al (0.0001–1 mmol/L) enhanced MDA+4-HDA formation in the synaptosomes. In addition, Al (1 mmol/L) raised protein carbonyl contents. Melatonin reduced, in a concentration-dependent manner, lipid and protein oxidation due to Al, FeCl₃ and ascorbic acid in the synaptosomal membranes. These results show that melatonin confers protection against Al-induced oxidative damage in synaptosomes and suggest that this indoleamine may be considered as a neuroprotective agent in Al toxicity because of its antioxidant activity.

Introduction

Melatonin is the main secretory product of the pineal gland, but its production is not limited to this organ [1], [2], [3], [4]. This molecule has a wide-ranging of endocrine, neural and immune functions in the body [5]. Recently intensive research has shown to be an effective antioxidant and free radical scavenger [6], [7].

Al is the third element in nature after oxygen and silicon. Humans are frequently exposed to Al because this metal is the third most abundant element in the Earth's crust [8] and it is a constituent of cooking utensils, medicines such as antiacids, deodorants and food additives [9]. In water, Al salts are frequently used as coagulants to remove colour and turbidity during water purification or treatment processes [10]. Patients with chronic renal failure undergoing long-term haemodialysis that use solutions with Al could accumulate the Al concentration in their brain and the other tissues [11]. Moreover, an augmentation of Al levels in the brain occurs physiologically with aging [12], [13]. Al disturbs certain cell signalling cascades [14] and induces neuronal apoptosis by its effect on the functioning of both the endoplasmic reticulum and mitochondria [15]. Its pro-oxidant effect in the brain [16] must be considered, because the nervous system is highly sensitive to oxidative damage resulting from free radicals since neurons have an elevated metabolic rate and the brain has poorly developed antioxidant protection mechanisms [17]. Al has been associated with the aetiology of Parkinson's disease and certain dementias, such as Alzheimer's disease, by high levels of Al in the brain in post-mortem evidences [10], [18], [19]. Al promotes the formation of amyloid-β protein plaques [20]. Although the mechanism of Al toxicity is not clearly known, it is reported than Al enhanced the activity of iron to cause oxidative damage [21], [22], [23], [24], [25].

Since melatonin is a powerful antioxidant, the aim of our study was to compare the effects of the indoleamine against experimentally Al-induced lipid and protein oxidation in synaptosomes, where free radicals are believed to be involved.

Section snippets

Chemicals

Melatonin, AlCl₃, FeCl₃, and ascorbic acid were purchased from Sigma (Madrid, Spain). The Bioxytech LPO-586 kit for lipid peroxidation was obtained from Caymen Chemical (Ann Arbor, MI, USA). Other chemicals used were of analytical grade and came from commercial sources. Melatonin was diluted in absolute ethanol and incubation buffer. Ethanol concentration was 2% (v/v) in the incubation volume. AlCl₃, FeCl₃, and ascorbic acid were diluted in the incubation buffer. All solutions were prepared

Effect of Al on synaptosomal lipid and protein oxidation

The incubation of synaptosomal membranes in the presence of FeCl₃ and ascorbic acid resulted in an augmentation of MDA+4-HDA levels and carbonyls contents in proteins indicating the oxidation of lipids and proteins, respectively, in the synaptosomes. The addition of AlCl₃ (0.0001, 0.001, 0.01, 0.1, 0.5 and 1 mmol/L) to the incubation buffer enhanced MDA+4-HDA levels and protein oxidation compared to those samples treated with FeCl₃ and ascorbic acid alone, with statistically significant

Discussion

Recently it has been proposed that oxidative stress is a feature of Alzheimer's disease and that Al could exacerbate oxidative damages [28]. Our studies show that Al promotes lipid and protein oxidation in synaptosomal membranes when this metal is combined with FeCl₃ and ascorbic acid. Al is a non-redox metal that does not initiate radical reactions. However, it is well documented that Al enhances in vitro iron-induced lipid peroxidation in biological membranes [22], [29], [30], [31]. Our data

Acknowledgements

This work was supported by the University of Zaragoza (Grant no. IBE2004B-B10-02), the Gobierno de Aragón (Aging and Oxidative Stress Physiology, Grant no. B40) and by F.I.S. from Instituto de Salud Carlos III (Grant no. G03/137). M.C.R. is a Predoctoral fellow from the Santander-Central-Hispano Bank and the University of Zaragoza, Spain.

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ToxicologyMelatonin reduces lipid and protein oxidative damage in synaptosomes due to aluminium

Abstract

Introduction

Section snippets

Chemicals

Effect of Al on synaptosomal lipid and protein oxidation

Discussion

Acknowledgements

Life Sci

Life Sci

J Inorg Biochem

Brain Res Bull

Exp Gerontol

Neurobiol Aging

Mol Aspects Med

J Inorg Biochem

Free Radic Biol Med

FEBS Lett

Brain Res

Biochim Biophys Acta

J Trace Elem Med Biol

Arch Biochem Biophys

Arch Biochem Biophys

Methods Enzymol

Anal Biochem

Free Radic Biol Med

Biochem Biophys Res Commun

Biochim Biophys Acta

Biochim Biophys Acta

Arch Biochem Biophys

Arch Biochem Biophys

Biochem Biophys Res Commun

Life Sci

Life Sci

FEBS Lett

Biochim Biophys Acta

Neuropharmacology

Toxicology
Melatonin reduces lipid and protein oxidative damage in synaptosomes due to aluminium