Elsevier

Brain Research

Volume 1008, Issue 2, 22 May 2004, Pages 245-251
Brain Research

Research report
Polyamines reduces lipid peroxidation induced by different pro-oxidant agents

https://doi.org/10.1016/j.brainres.2004.02.036Get rights and content

Abstract

Polyamines, among other functions, are considered to act as a free radical scavenger and antioxidant. The quinolinic acid (QA), sodium nitroprusside (SNP) and iron (Fe+2) stimulate production of free radicals and lipid peroxidation. In the present study, we investigated the free radical and/or aldehyde scavenger effects of polyamines spermine and spermidine on thiobarbituric acid reactive species (TBARS) production induced by QA, SNP, Fe+2/EDTA system and free Fe2+ in rat brain. Spermine and spermidine inhibited QA-induced TBARS production; however spermine was a better antioxidant than spermidine. Spermine also inhibited SNP-, Fe+2/EDTA- and free Fe2+-induced TBARS production, but had a modest effect. Spermidine, in turn, also discreetly inhibited SNP-, Fe+2/EDTA- and free Fe2+-induced TBARS production. In the presence of MK-801, QA-induced TBARS production was considerably more inhibited by polyamines. In addition, arcaine does not affect the reducer effect of polyamines. The present findings suggest that the observed effects of polyamines are not related to the activation of NMDA receptor but with their antioxidant and free radical scavenger properties.

Introduction

Free radicals are now accepted as important mediators of tissue injury in several neurodegenerative models [6], [22], [28], [45], [49]. The brain is particularly susceptible to free radical damage because of its high utilization of oxygen and its relatively low concentration of antioxidants enzymes and free radicals scavengers. These free radicals may attack membrane lipids, proteins and nucleic acids to cause cell damage or death [24], [25], [36], [37]. During oxidative stress, cell injury can be amplified by reactive compounds that are by-products of oxidative damage including malondialdehyde, 4-hydroxyalkenals and numerous 2-alkenals [17]. Based on the observation of elevated levels of the aldehydes and/or their respective macromolecular adducts, these lipid peroxidation products appear to contribute to the etiology of a number of chronic diseases including neurodegenerative conditions, chronic inflammatory diseases, alcoholic liver disease, cardiovascular disorders and diabetic complications [3], [10], [13], [15], [54].

Pharmacological efforts to attenuate oxidative injury in degenerative diseases have typically focused on drugs with antioxidant properties [48]. Such approaches provide a ‘first line of defense’ against free radicals, but do not target secondary products of oxidative stress, including numerous reactive α,β-unsaturated aldehydes. A complementary strategy involves identification of low-molecular weight drugs possessing nucleophilic centers (e.g. primary amine groups) that exhibit high reactivity toward endogenous aldehydes. In biological systems such drugs might act as ‘aldehyde scavengers’, sparing cellular constituents and slowing disease progression [46].

The polyamines (spermine, spermidine and putrescine) are ornitine metabolic products, which are found at high concentrations in the brain [2], [57]. The primary and secondary amine moieties of polyamines always carry a charge at physiological pH, resulting in low molecular weight ‘organic cations’, the most important characteristic required for ‘aldehyde scavengers’. This polycationic quality has attracted the attention of researchers and led to the hypothesis that polyamines could affect physiological systems by binding to anionic sites, such as those associated with nucleic acids and membrane phospholipids [27]. These amines are involved with numerous cellular functions, including free radical scavenger, antioxidant and antiinflammatory properties [1], [14], [16], [19], [23], [32]. In addition, recent experimental evidence supports a role for polyamines as transcriptional regulators in the nucleus of neural eukaryotic cells [30]. However, the precise mechanisms by which polyamines exert their regulatory effects are unresolved. It have been reported that spermine inhibits both NADPH- and ascorbic acid-dependent lipid peroxidation in liver microsomes [29].

Quinolinic acid (QA) is a neuroactive metabolite of the tryptophan-kinurenine pathway which can be produced by macrophages and microglia [11], [51]. It is present in both the human and rat brain [58] and it has been implicated in the pathogenesis of a variety of human neurological diseases [7], [31]. QA is recognized pharmacologically as an endogenous glutamate agonist with a relative selectivity for the NMDA receptor in the brain [20]. Since it is not readily metabolized in the synaptic cleft, it stimulates the NMDA receptor for prolonged periods. This sustained stimulation results in opening of calcium channels causing Ca+2 influx followed by Ca+2-dependent enhancement of free radical production leading to molecular damage and often to cell death [9], [52]. In addition, it has been reported that QA was able to stimulate lipid peroxidation in rat brain homogenates [39], [43]. These findings have suggested the involvement of free radicals and oxidative stress in the pattern of toxicity elicited by QA in the nervous system [41].

Sodium nitroprusside (SNP) has been suggested to cause cytotoxicity through the release of cyanide and/or nitric oxide (NO) [4], [5]. There is a growing number of recent studies concerning the role of NO, a molecule that is regarded as universal neuronal messenger in the central nervous system, in the pathophysiology of such disorders as Alzheimer's and Parkinson's diseases, stroke, trauma, seizure disorders, etc. [8], [12], [34], [55]. NO is a free radical with short half-life (<30 s). Although NO acts independently, it also may cause neuronal damage in cooperation with other reactive oxygen species (ROS) [26], [35].

Therefore, the aim of this study was to verify the potential protective effects of spermine and spermidine against thiobarbituric reactive species (TBARS) production induced by different pro-oxidant agents and provide an hypothesis of the mechanisms by which polyamines exert their regulatory effects.

Section snippets

Chemicals

Tris(hydroximethyl)aminomethane, spermine, spermidine, arcaine, quinolinic acid, thiobarbituric acid and malonaldehyde bis-(dimethyl acetal) (MDA) were obtained from Sigma (St. Louis, MO, USA). MK-801 was obtained from Research Biochemicals International (Natick, MA, USA). Sodium nitroprusside was obtained from Merck (Darmstadt, Germany). Ethylenediaminetetraacetic (EDTA), HCl and acetic acid were obtained from Merck (Rio de Janeiro, RJ, Brazil). Ammonium iron(II) sulfate was obtained from

Quinolinic acid vs. Polyamines

Spermine (0.000033–3.33 μM) did not change the basal production of TBARS by brain homogenates. QA elicited a marked increase in TBARS production and this effect was not modified by spermine at the same concentrations range (data not shown). Similarly, spermidine (0.00003–0.002 μM) did not change the basal production of TBARS. QA caused a marked increase in TBARS production which was not counteracted by these concentrations of spermidine (data not shown).

At higher concentrations tested, spermine

Discussion

The main findings of the present study can be summarized in the following topics: (1) spermine and spermidine reduced QA-induced lipid peroxidation; (2) spermine and spermidine, at relatively high concentrations, have a modest inhibitory effect against sodium nitroprusside-induced lipid peroxidation; and (3) at relatively high concentrations, both amines have a modest effect as inhibitors of Fe2+- or Fe2+/EDTA-induced TBARS production.

Taken together, these results can indicate that the

Acknowledgements

This study was supported by CNPq (474241/2003-3, 523761/95-3 and 500096/2003-1), CAPES and FAPERGS (01/1338.9).

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