Optimization of a microchip electrophoresis method with electrochemical detection for the determination of nitrite in macrophage cells as an indicator of nitric oxide production

Siegel, Joseph Michael; Schilly, Kelci M.; Wijesinghe, Manjula B.; Caruso, Giuseppe; Fresta, Claudia G.; Lunte, Susan M.

doi:10.1039/c8ay02014k

Cited by 19 publications

(14 citation statements)

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“…Oxidative stress and neuroinflammation play a significant role in the pathogenesis of depression (Bhattacharya and Drevets, 2017;Caruso et al, 2019a) and AD (Huang et al, 2016;Knezevic and Mizrahi, 2018). During the inflammation process, both inducible nitric oxide synthase (iNOS), responsible for nitric oxide production (Aktan, 2004;Metto et al, 2013), and NADPH oxidase 2 (Nox2), responsible for superoxide production (de Campos et al, 2015), are overactivated in immune cells including microglia (Siegel et al, 2019). When the above-mentioned enzymes are simultaneously activated, they synergistically promote neuronal cell death by generating peroxynitrite (Beckman and Crow, 1993).…”

Section: Fluoxetine and Vortioxetine Decreased The Expression Of Inos And Nox2 Mrnasmentioning

confidence: 99%

Antioxidant Activity of Fluoxetine and Vortioxetine in a Non-Transgenic Animal Model of Alzheimer’s Disease

et al. 2021

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Depression is a risk factor for the development of Alzheimer’s disease (AD). A neurobiological and clinical continuum exists between AD and depression, with neuroinflammation and oxidative stress being involved in both diseases. Second-generation antidepressants, in particular selective serotonin reuptake inhibitors (SSRIs), are currently investigated as neuroprotective drugs in AD. By employing a non-transgenic AD model, obtained by intracerebroventricular (i.c.v.) injection of amyloid-β (Aβ) oligomers in 2-month-old C57BL/6 mice, we recently demonstrated that the SSRI fluoxetine (FLX) and the multimodal antidepressant vortioxetine (VTX) reversed the depressive-like phenotype and memory deficits induced by Aβ oligomers rescuing the levels of transforming growth factor-β1 (TGF-β1). Aim of our study was to test FLX and VTX for their ability to prevent oxidative stress in the hippocampus of Aβ-injected mice, a brain area strongly affected in both depression and AD. The long-term intraperitoneal (i.p.) administration of FLX (10 mg/kg) or VTX (5 and 10 mg/kg) for 24 days, starting 7 days before Aβ injection, was able to prevent the over-expression of inducible nitric oxide synthase (iNOS) and NADPH oxidase 2 (Nox2) induced by Aβ oligomers. Antidepressant pre-treatment was also able to rescue the mRNA expression of glutathione peroxidase 1 (Gpx1) antioxidant enzyme. FLX and VTX also prevented Aβ-induced neurodegeneration in mixed neuronal cultures treated with Aβ oligomers. Our data represent the first evidence that the long-term treatment with the antidepressants FLX or VTX can prevent the oxidative stress phenomena related to the cognitive deficits and depressive-like phenotype observed in a non-transgenic animal model of AD.

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Section: Fluoxetine and Vortioxetine Decreased The Expression Of Inos And Nox2 Mrnasmentioning

confidence: 99%

Antioxidant Activity of Fluoxetine and Vortioxetine in a Non-Transgenic Animal Model of Alzheimer’s Disease

et al. 2021

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“…Peroxynitrite, the reaction product between NO and O 2 −• , can react and consequently damage fatty acids, proteins, DNA, and mitochondria leading to oxidative and nitrosative stress [22], inflammation [23], and then neurodegenerative phenomena [24,25]. During the inflammation process, both inducible nitric oxide synthase (iNOS), responsible for NO production [26,27], and NADPH oxidase (Nox), responsible for O 2 −• production [28], are overactivated in immune cells such as macrophage and microglia [20,29,30]. When simultaneously activated, iNOS and Nox act synergistically to promote neuronal death through the generation of peroxynitrite, which is a more dangerous species if compared with NO and/or O 2 −• [31].…”

Section: Introductionmentioning

confidence: 99%

Carnosine Prevents Aβ-Induced Oxidative Stress and Inflammation in Microglial Cells: A Key Role of TGF-β1

Caruso

Fresta

Musso

et al. 2019

Cells

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Carnosine (β-alanyl-L-histidine), a dipeptide, is an endogenous antioxidant widely distributed in excitable tissues like muscles and the brain. Carnosine is involved in cellular defense mechanisms against oxidative stress, including the inhibition of amyloid-beta (Aβ) aggregation and the scavenging of reactive species. Microglia play a central role in the pathogenesis of Alzheimer’s disease, promoting neuroinflammation through the secretion of inflammatory mediators and free radicals. However, the effects of carnosine on microglial cells and neuroinflammation are not well understood. In the present work, carnosine was tested for its ability to protect BV-2 microglial cells against oligomeric Aβ1-42-induced oxidative stress and inflammation. Carnosine prevented cell death in BV-2 cells challenged with Aβ oligomers through multiple mechanisms. Specifically, carnosine lowered the oxidative stress by decreasing NO and O2−• intracellular levels as well as the expression of iNOS and Nox enzymes. Carnosine also decreased the secretion of pro-inflammatory cytokines such as IL-1β, simultaneously rescuing IL-10 levels and increasing the expression and the release of TGF-β1. Carnosine also prevented Aβ-induced neurodegeneration in mixed neuronal cultures challenged with Aβ oligomers, and these neuroprotective effects were completely abolished by SB431542, a selective inhibitor of the type-1 TGF-β receptor. Our data suggest a multimodal mechanism of action of carnosine underlying its protective effects on microglial cells against Aβ toxicity with a key role of TGF-β1 in mediating these protective effects.

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“…When inflammation processes take place, inducible nitric oxide synthase (iNOS), responsible for NO production [15], as well as NADPH oxidase (Nox), responsible for O 2 − • production [16], are overexpressed in immune cells such as macrophages and microglia [17,18]. Different types of cells are involved in the immune response, among others macrophages are those primarily activated [19,20].…”

Section: Introductionmentioning

confidence: 99%

Carnosine Decreases PMA-Induced Oxidative Stress and Inflammation in Murine Macrophages

et al. 2019

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Carnosine is an endogenous dipeptide composed of β-alanine and L-histidine. This naturally occurring molecule is present at high concentrations in several mammalian excitable tissues such as muscles and brain, while it can be found at low concentrations in a few invertebrates. Carnosine has been shown to be involved in different cellular defense mechanisms including the inhibition of protein cross-linking, reactive oxygen and nitrogen species detoxification as well as the counteraction of inflammation. As a part of the immune response, macrophages are the primary cell type that is activated. These cells play a crucial role in many diseases associated with oxidative stress and inflammation, including atherosclerosis, diabetes, and neurodegenerative diseases. In the present study, carnosine was first tested for its ability to counteract oxidative stress. In our experimental model, represented by RAW 264.7 macrophages challenged with phorbol 12-myristate 13-acetate (PMA) and superoxide dismutase (SOD) inhibitors, carnosine was able to decrease the intracellular concentration of superoxide anions (O2−•) as well as the expression of Nox1 and Nox2 enzyme genes. This carnosine antioxidant activity was accompanied by the attenuation of the PMA-induced Akt phosphorylation, the down-regulation of TNF-α and IL-6 mRNAs, and the up-regulation of the expression of the anti-inflammatory mediators IL-4, IL-10, and TGF-β1. Additionally, when carnosine was used at the highest dose (20 mM), there was a generalized amelioration of the macrophage energy state, evaluated through the increase both in the total nucleoside triphosphate concentrations and the sum of the pool of intracellular nicotinic coenzymes. Finally, carnosine was able to decrease the oxidized (NADP+)/reduced (NADPH) ratio of nicotinamide adenine dinucleotide phosphate in a concentration dependent manner, indicating a strong inhibitory effect of this molecule towards the main source of reactive oxygen species in macrophages. Our data suggest a multimodal mechanism of action of carnosine underlying its beneficial effects on macrophage cells under oxidative stress and inflammation conditions.

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Optimization of a microchip electrophoresis method with electrochemical detection for the determination of nitrite in macrophage cells as an indicator of nitric oxide production

Abstract: A transient isotachophoresis separation and Pt black working electrode allow detection of nitrite in stimulated macrophage cells with improved sensitivity.

Cited by 19 publications

References 53 publications

Antioxidant Activity of Fluoxetine and Vortioxetine in a Non-Transgenic Animal Model of Alzheimer’s Disease

Antioxidant Activity of Fluoxetine and Vortioxetine in a Non-Transgenic Animal Model of Alzheimer’s Disease

Carnosine Prevents Aβ-Induced Oxidative Stress and Inflammation in Microglial Cells: A Key Role of TGF-β1

Carnosine Decreases PMA-Induced Oxidative Stress and Inflammation in Murine Macrophages

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