In vitro identification of mitochondrial oxidative stress production by time-resolved fluorescence imaging of glioma cells

Tomkova, Silvia; Misuth, Matus; Lenkavská, Lenka; Miškovský, Pavol; Huntošová, Veronika

doi:10.1016/j.bbamcr.2018.01.012

Cited by 22 publications

(30 citation statements)

References 74 publications

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“…Microscopy with Mitotracker orange (stains mitochondria in live cells) was performed with control, menadione-treated (10 µM) and picein-treated (25 µM) samples after menadione exposure. The live cell imaging of SH-SY5Y cells after menadione exposure showed low emission of Mitotracker fluorescence, could be indicating an overproduction of ROS, as also reported by Tomkova et al [45]. Moreover, the qualitative observations from Figure 7 clearly reveal that the menadione-exposed cells were morphologically distinct with structural abnormalities, and such deviations could be interpreted as mitochondrial dysfunction, as has been reported by other researchers [46,47].…”

Section: Live Cell Imagingsupporting

confidence: 83%

“…MitoSOX red, a novel fluorogenic probe, can bind to the nuclear DNA and therefore identify superoxide in the live cells of mitochondria [64]. In addition to this, the selective mitochondrial probe Mitotracker orange was employed in this study, which is highly receptive to mitochondrial membrane potential and considered a sensitive indicator for the measurement of oxidative stress in mitochondria [45]. The accumulation of Mitotracker in cells mainly depends on the mitochondrial membrane potential [68] because it disseminates throughout the plasma membrane and accumulates in live mitochondria.…”

Section: Discussionmentioning

confidence: 99%

“…The notation for picein and gastrodin were LogP −0.43 and −0.99, topological polar surface area 116. 45 Figure 2. CHIPPI web server was used for the network protein-protein interaction (PPI) analysis.…”

Section: Testing Of Lipinski's Rule Of 5 and Docking Analysismentioning

confidence: 99%

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Plant-Derived Natural Biomolecule Picein Attenuates Menadione Induced Oxidative Stress on Neuroblastoma Cell Mitochondria

et al. 2020

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Several bioactive compounds are in use for the treatment of neurodegenerative disorders, such as Alzheimer’s and Parkinson’s disease. Historically, willow (salix sp.) bark has been an important source of salisylic acid and other natural compounds with anti-inflammatory, antipyretic and analgesic properties. Among these, picein isolated from hot water extract of willow bark, has been found to act as a natural secondary metabolite antioxidant. The aim of this study was to investigate the unrevealed pharmacological action of picein. In silico studies were utilized to direct the investigation towards the neuroprotection abilities of picein. Our in vitro studies demonstrate the neuroprotective properties of picein by blocking the oxidative stress effects, induced by free radical generator 2-methyl-1,4-naphthoquinone (menadione, MQ), in neuroblastoma SH-SY5Y cells. Several oxidative stress-related parameters were evaluated to measure the protection for mitochondrial integrity, such as mitochondrial superoxide production, mitochondrial activity (MTT), reactive oxygen species (ROS) and live-cell imaging. A significant increase in the ROS level and mitochondrial superoxide production were measured after MQ treatment, however, a subsequent treatment with picein was able to mitigate this effect by decreasing their levels. Additionally, the mitochondrial activity was significantly decreased by MQ exposure, but a follow-up treatment with picein recovered the normal metabolic activity. In conclusion, the presented results demonstrate that picein can significantly reduce the level of MQ-induced oxidative stress on mitochondria, and thereby plays a role as a potent neuroprotectant.

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Section: Live Cell Imagingsupporting

confidence: 83%

Section: Discussionmentioning

confidence: 99%

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Plant-Derived Natural Biomolecule Picein Attenuates Menadione Induced Oxidative Stress on Neuroblastoma Cell Mitochondria

et al. 2020

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“…Briefly, the OCR was measured by the sequential addition of oligomycin (an ATP synthase inhibitor, 1 μM), phenylhydrazone (FCCP, a mitochondrial respiration uncoupler, 0.5 μM), and rotenone (a complete respiratory inhibitor, 2 μM). This protocol is normally used for the determination of basal OCR, ATP‐linked respiration, and maximal respiration (Tomkova, Misuth, Lenkavska, Miskovsky, & Huntosova, ). After the measurement of mitochondrial respiratory capacity, the cells were lysed, and the total protein was quantified using the BCA protein assay.…”

Section: Methodsmentioning

confidence: 99%

Roflupram exerts neuroprotection via activation of CREB/PGC‐1α signalling in experimental models of Parkinson's disease

Zhong

Dong

Qin

et al. 2020

British J Pharmacology

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Background and Purpose:Roflupram improves cognition and limits neuroinflammation in the brain. However, the beneficial effects of roflupram on Parkinson's disease (PD) remain unknown. Therefore, we aimed to elucidate the pharmacological effects and mechanisms of action of ROF in experimental models of PD. Experimental Approach:We used an in vitro PD model of SH-SY5Y cells exposed to 1-methyl-4-phenylpyridinium iodide (MPP + ). Cell viability and apoptosis were analysed via the MTT assay and flow cytometry. Mitochondrial morphology, mitochondrial respiratory capacity, and ROS were measured by a mitochondrial tracker, Seahorse Analyzer, and a MitoSOX-Red dye. For in vivo PD model, behavioural tests, Nissl staining, and immunohistochemistry were used to evaluate protection by roflupram. The levels of TH, cAMP response element-binding protein (CREB), and PPARγ coactivator-1α (PGC-1α) were analysed by western blotting. Key Results: Roflupram decreased MPP + -induced apoptosis in SH-SY5Y cells and human dopaminergic neurons. Roflupram also increased mitochondrial respiratory capacity, decreased ROS production, and restored mitochondrial morphology. Roflupram reversed the MPP + -induced reductions of phosphorylated CREB, PGC-1α and TH. These protective effects were blocked by the PKA inhibitor H-89 or by PGC-1α siRNA. In mice treated with MPTP, roflupram significantly improved motor functions. Roflupram prevented both dopaminergic neuronal loss and the reduction of phosphorylated CREB and PGC-1α in the substantia nigra and striatum. Conclusion and Implications: Roflupram protected dopaminergic neurons from apoptosis via the CREB/PGC-1α pathway in PD models. Hence, roflupram has potential as a protective drug in the treatment of PD.Abbreviations: CREB, cAMP response element-binding protein; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; MMP, mitochondrial membrane potential; MPP + , 1-methyl-4-phenylpyridinium iodide; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; MPTP, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; OCR, oxygen consumption rate; PD, Parkinson's disease; PGC-1α, PPARγ coactivator-1α; SN, substantia nigra; TMRE, tetramethylrhodamine ethyl ester perchlorate.Jiahong Zhong and Wenli Dong have equal contribution to this work.

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“…Myelin is not an inert molecule that simply coats the axon but rather a metabolically active structure: in fact, myelin and the neuronal axon must be regarded as a single metabolically combined functional unit. Oligodendrocyte creates myelin channels—located at the end of the axon—that allow for lipid macromolecules to be transported and moved [20].…”

Section: Introductionmentioning

confidence: 99%

Myelin Disturbances Produced by Sub-Toxic Concentration of Heavy Metals: The Role of Oligodendrocyte Dysfunction

Maiuolo

Macrì

Bava

et al. 2019

IJMS

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Evidence has been accumulated demonstrating that heavy metals may accumulate in various organs, leading to tissue damage and toxic effects in mammals. In particular, the Central Nervous System (CNS) seems to be particularly vulnerable to cumulative concentrations of heavy metals, though the pathophysiological mechanisms is still to be clarified. In particular, the potential role of oligodendrocyte dysfunction and myelin production after exposure to subtoxic concentration I confirmed. It is ok of heavy metals is to be better assessed. Here we investigated on the effect of sub-toxic concentration of several essential (Cu2 +, Cr3 +, Ni2 +, Co2+) and non-essential (Pb2 +, Cd2+, Al3+) heavy metals on human oligodendrocyte MO3.13 and human neuronal SHSY5Y cell lines (grown individually or in co-culture). MO3.13 cells are an immortal human–human hybrid cell line with the phenotypic characteristics of primary oligodendrocytes but following the differentiation assume the morphological and biochemical features of mature oligodendrocytes. For this reason, we decided to use differentiated MO3.13 cell line. In particular, exposure of both cell lines to heavy metals produced a reduced cell viability of co-cultured cell lines compared to cells grown separately. This effect was more pronounced in neurons that were more sensitive to metals than oligodendrocytes when the cells were grown in co-culture. On the other hand, a significant reduction of lipid component in cells occurred after their exposure to heavy metals, an effect accompanied by substantial reduction of the main protein that makes up myelin (MBP) in co-cultured cells. Finally, the effect of heavy metals in oligodendrocytes were associated to imbalanced intracellular calcium ion concentration as measured through the fluorescent Rhod-2 probe, thus confirming that heavy metals, even used at subtoxic concentrations, lead to dysfunctional oligodendrocytes. In conclusion, our data show, for the first time, that sub-toxic concentrations of several heavy metals lead to dysfunctional oligodendrocytes, an effect highlighted when these cells are co-cultured with neurons. The pathophysiological mechanism(s) underlying this effect is to be better clarified. However, imbalanced intracellular calcium ion regulation, altered lipid formation and, finally, imbalanced myelin formation seem to play a major role in early stages of heavy metal-related oligodendrocyte dysfunction.

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In vitro identification of mitochondrial oxidative stress production by time-resolved fluorescence imaging of glioma cells

Cited by 22 publications

References 74 publications

Plant-Derived Natural Biomolecule Picein Attenuates Menadione Induced Oxidative Stress on Neuroblastoma Cell Mitochondria

Plant-Derived Natural Biomolecule Picein Attenuates Menadione Induced Oxidative Stress on Neuroblastoma Cell Mitochondria

Roflupram exerts neuroprotection via activation of CREB/PGC‐1α signalling in experimental models of Parkinson's disease

Myelin Disturbances Produced by Sub-Toxic Concentration of Heavy Metals: The Role of Oligodendrocyte Dysfunction

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