Creatine affords protection against glutamate-induced nitrosative and oxidative stress

Abstract

Creatine has been reported to exert beneficial effects in several neurodegenerative diseases in which glutamatergic excitotoxicity and oxidative stress play an etiological role. The purpose of this study was to investigate the protective effects of creatine, as compared to the N-Methyl-d-Aspartate (NMDA) receptor antagonist dizocilpine (MK-801), against glutamate or hydrogen peroxide (H2O2)-induced injury in human neuroblastoma SH-SY5Y cells. Exposure of cells to glutamate (60-80 mM) or H2O2 (200-300 μM) for 24 h decreased cellular viability and increased dichlorofluorescein (DCF) fluorescence (indicative of increased reactive oxygen species, ROS) and nitric oxide (NO) production (assessed by mono-nitrogen oxides, NOx, levels). Creatine (1-10 mM) or MK-801 (0.1-10 μM) reduced glutamate- and H2O2-induced toxicity. The protective effect of creatine against glutamate-induced toxicity involves its antioxidant effect, since creatine, similar to MK-801, prevented the increase on DCF fluorescence induced by glutamate or H2O2. Furthermore, creatine or MK-801 blocked glutamate- and H2O2-induced increases in NOx levels. In another set of experiments, the repeated, but not acute, administration of creatine (300 mg/kg, po) in mice prevented the decreases on cellular viability and mitochondrial membrane potential (assessed by tetramethylrhodamine ethyl ester, TMRE, probe) of hippocampal slices incubated with glutamate (10 mM). Creatine concentration-dependent decreased the amount of nitrite formed in the reaction of oxygen with NO produced from sodium nitroprusside solution, suggesting that its protective effect against glutamate or H2O2-induced toxicity might be due to its scavenger activity. Overall, the results suggest that creatine may be useful as adjuvant therapy for neurodegenerative disease treatments.

Gateways for Glutamate Neuroprotection in Parkinson’s Disease (PD): Essential Role of EAAT3 and NCX1 Revealed in an In Vitro Model of PD

“…Experimental settings used in our work to explore and characterize the potential of glutamate-driven metabolism to limit PD-like degeneration were chosen to minimize excitotoxic neuronal damage, as discussed below. In our model glutamate, in terms of both concentration and time of exposure, had no evident toxic effects under control conditions, which is consistent with previous studies based on SH-SY5Y cells, where higher concentrations of glutamate (up to tens of millimolar levels) and/or longer exposure (within hours-to-days time frames) were reported to significantly impair cell functions/viability [ 91 , 92 , 93 , 94 , 95 ]. Several lines of evidence converge to support that the metabolic fate of glutamate in brain cells is highly influenced by the glutamate levels in the extracellular milieu: glutamate metabolism that feeds ATP-generating pathways is typically favored when neuronal (including mesencephalic cell preparations) or glial cultures are exposed for 1–2 h to glutamate concentrations in the hundreds of micromolar range [ 40 , 89 , 96 , 97 , 98 ].…”

Metabolomic Responses to Manganese Dose in SH-SY5Y Human Neuroblastoma Cells

“…Cunha et al, demonstrated protective effects of creatine on H 2 O 2 -induced toxicity in SH-SY5Y cells (Cunha et al, 2016). On similar lines, we performed analysis in SH-SY5Y cells to determine whether creatine has protective roles in toxic Mn dosecaused alterations in energy and neurotransmitter metabolites using HRM.…”

“…Creatine and phosphocreatine are shown to counteract apoptosis, excitotoxicity, and oxidative stress (Lawler et al, 2002;O'Gorman et al, 1997;Xu et al, 1996). Creatine has also been reported to be a neuroprotective agent (Genius et al, 2012;Klein and Ferrante, 2007) and its therapeutic use has been proposed for neurological disorder such as Parkinson's, Alzheimer's, ALS, HD, and mitochondrial diseases (Cunha et al, 2016;Klein and Ferrante, 2007;Klivenyi et al, 1999;Lin et al, 2013;Rae and Broer, 2015;Stevens et al, 2014). Consistently, we also found a protective effect of creatine on toxicological Mn dose-induced metabolic response in SH-SY5Y cells (see Supplementary Figure 2) suggesting that cellular creatine level plays an important role in metabolic responses to Mn doses.…”

Metabolic responses of BV‐2 cells to puerarin on its polarization using ultra‐performance liquid chromatography–mass spectrometry

“…It is a potent antioxidant and ameliorates glutamatergic excitotoxicity and apoptosis in vitro and in vivo. Therefore, creatine supplementation may offer some utility for treating neurodegenerative diseases (Cunha et al, ). Mitochondria are not only fundamental to cellular bioenergetics, but are also key mediators of apoptosis and can be linked directly or indirectly to other deleterious processes involved in neurodegeneration (Beal, ).…”