β-Lapachone Induces Acute Oxidative Stress in Rat Primary Astrocyte Cultures that is Terminated by the NQO1-Inhibitor Dicoumarol

Steinmeier, Johann; Kube, Sophie; Karger, Gabriele; Ehrke, Eric; Dringen, Ralf

doi:10.1007/s11064-020-03104-0

Cited by 11 publications

(19 citation statements)

References 60 publications

(108 reference statements)

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“…However, the generated hydroquinone is labile and autoxidises quickly in two distinct one-electron steps, thereby generating superoxide which can subsequently damage tumour cells that are known to express high activities of NQO1 [ 5 ]. However, as also brain astrocytes contain substantial activity of NQO1 [ 6 ] such normal body cells have to be considered as potential unwanted target of a β-lapachone treatment [ 7 ].…”

Section: Introductionmentioning

confidence: 99%

“…In addition, submicromolar concentrations of β-lapachone have been shown to activate glutamate dehydrogenase and to counteract iodoacetate induced toxicity in cultured astrocytes [ 15 ]. In contrast, in concentrations above 10 µM β-lapachone causes acute ROS formation, glutathione disulfide (GSSG) accumulation, metabolic impairment and toxicity in cultured astrocytes [ 7 ]. All these adverse effects of a treatment with high micromolar concentrations of β-lapachone are abolished in the presence of the NQO1 inhibitor dicoumarol, demonstrating that the reduction of β-lapachone by NQO1 is exclusively responsible to initiate the adverse consequences of an acute β-lapachone treatment [ 7 ].…”

Section: Introductionmentioning

confidence: 99%

“…This allows the investigation of intracellular metabolic pathways that can provide NAD(P)H for the NQO1 reduction by monitoring extracellular WST1 reduction. Menadione has been used as redox cycler for such studies [ 6 , 17 ], but also β-lapachone has the potential to mediate as redox cycler NQO1-dependent extracellular WST1 formazan generation [ 7 ]. However, both redox cyclers have been reported in higher concentrations to cause oxidative stress in astrocytes [ 7 , 18 , 19 ].…”

Section: Introductionmentioning

confidence: 99%

“…Menadione has been used as redox cycler for such studies [ 6 , 17 ], but also β-lapachone has the potential to mediate as redox cycler NQO1-dependent extracellular WST1 formazan generation [ 7 ]. However, both redox cyclers have been reported in higher concentrations to cause oxidative stress in astrocytes [ 7 , 18 , 19 ].…”

Section: Introductionmentioning

confidence: 99%

“…β-lapachone in a concentration of 20 µM has recently also been reported to act in cultured astrocytes as electron cycler that transfers in its reduced β-lapachol form electrons derived from intracellular NQO1-dependent reduction through the membrane to allow extracellular WST1 reduction [ 7 ]. However, such high concentrations of β-lapachone induce oxidative stress [ 7 ] which has been demonstrated to affect metabolic pathways such as glycolysis and pentose-phosphate pathway which contribute to the regeneration of NADH and NADPH [ 7 , 19 – 21 ].…”

Section: Introductionmentioning

confidence: 99%

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β-lapachone-mediated WST1 Reduction as Indicator for the Cytosolic Redox Metabolism of Cultured Primary Astrocytes

Watermann

Dringen

2023

Neurochem Res

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Electron cycler-mediated extracellular reduction of the water-soluble tetrazolium salt 1 (WST1) is frequently used as tool for the determination of cell viability. We have adapted this method to monitor by determining the extracellular WST1 formazan accumulation the cellular redox metabolism of cultured primary astrocytes via the NAD(P)H-dependent reduction of the electron cycler β-lapachone by cytosolic NAD(P)H:quinone oxidoreductase 1 (NQO1). Cultured astrocytes that had been exposed to β-lapachone in concentrations of up to 3 µM remained viable and showed an almost linear extracellular accumulation of WST1 formazan for the first 60 min, while higher concentrations of β-lapachone caused oxidative stress and impaired cell metabolism. β-lapachone-mediated WST1 reduction was inhibited by the NQO1 inhibitors ES936 and dicoumarol in a concentration-dependent manner, with half-maximal inhibition observed at inhibitor concentrations of about 0.3 µM. β-lapachone-mediated WST1 reduction depended strongly on glucose availability, while mitochondrial substrates such as lactate, pyruvate or ketone bodies allowed only residual β-lapachone-mediated WST1 reduction. Accordingly, the mitochondrial respiratory chain inhibitors antimycin A and rotenone hardly affected astrocytic WST1 reduction. Both NADH and NADPH are known to supply electrons for reactions catalysed by cytosolic NQO1. Around 60% of the glucose-dependent β-lapachone-mediated WST1 reduction was prevented by the presence of the glucose-6-phosphate dehydrogenase inhibitor G6PDi-1, while the glyceraldehyde-3-phosphate dehydrogenase inhibitor iodoacetate had only little inhibitory potential. These data suggest that pentose phosphate pathway-generated NADPH, and not glycolysis-derived NADH, is the preferred electron source for cytosolic NQO1-catalysed reductions in cultured astrocytes.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

β-lapachone-mediated WST1 Reduction as Indicator for the Cytosolic Redox Metabolism of Cultured Primary Astrocytes

Watermann

Dringen

2023

Neurochem Res

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Tumor Microenvironment Triggered Cascade‐Activation Nanoplatform for Synergistic and Precise Treatment of Hepatocellular Carcinoma

Xin

Cai

et al. 2021

Adv Healthcare Materials

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Hepatocellular carcinoma (HCC) is one of the most common and deadliest malignancy cancers, which remains a major global health problem. At present, over 50% of patients with HCC have implemented systemic therapies, such as interventional therapy or local chemotherapy that are scarcely effective and induce serious side effects to the remaining normal liver, further limiting their clinical outcomes. Herein, a tumor microenvironment triggered cascade-activation nanoplatform (A-NP Lap/TPZ ) is prepared based on -lapachone ( -Lap) and tirapazamine (TPZ) for the synergistic therapy of HCC. The A-NP Lap/TPZ exerts its targeting effect by binding to the receptor of tumor cells with an external aptamer. In the tumor microenvironment, the nanoplatform can realize H 2 O 2 -triggered disassembly to release -Lap and TPZ. The released -Lap generates ROS to induce tumor cell apoptosis under the catalysis of the tumor cell over-expressed NAD(P)H-quinone oxidoreductase-1 (NQO1) enzyme. In this process, oxygen is consumed to intensify tumor hypoxia, and eventually cascade activates TPZ to exert the anti-tumor effect. The studies in vitro and in vivo consistently demonstrate that the as-prepared A-NP Lap/TPZ nanoplatform possesses an excellent synergistic anti-tumor effect. This design of nanoplatform with cascade activation effect provides a promising strategy for HCC treatment.

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Consumption and Metabolism of Extracellular Pyruvate by Cultured Rat Brain Astrocytes

et al. 2022

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Brain astrocytes are considered as glycolytic cell type, but these cells also produce ATP via mitochondrial oxidative phosphorylation. Exposure of cultured primary astrocytes in a glucose-free medium to extracellular substrates that are known to be metabolised by mitochondrial pathways, including pyruvate, lactate, beta-hydroxybutyrate, alanine and acetate, revealed that among the substrates investigated extracellular pyruvate was most efficiently consumed by astrocytes. Extracellular pyruvate was consumed by the cells almost proportional to time over hours in a concentration-dependent manner with apparent Michaelis–Menten kinetics [Km = 0.6 ± 0.1 mM, Vmax = 5.1 ± 0.8 nmol/(min × mg protein)]. The astrocytic consumption of pyruvate was strongly impaired in the presence of the monocarboxylate transporter 1 (MCT1) inhibitor AR-C155858 or by application of a 10-times excess of the MCT1 substrates lactate or beta-hydroxybutyrate. Pyruvate consumption by viable astrocytes was inhibited in the presence of UK5099, an inhibitor of the mitochondrial pyruvate carrier, or after application of the respiratory chain inhibitor antimycin A. In contrast, the mitochondrial uncoupler BAM15 strongly accelerated cellular pyruvate consumption. Lactate and alanine accounted after 3 h of incubation with pyruvate for around 60% and 10%, respectively, of the pyruvate consumed by the cells. These results demonstrate that consumption of extracellular pyruvate by astrocytes involves uptake via MCT1 and that the velocity of pyruvate consumption is strongly modified by substances that affect the entry of pyruvate into mitochondria or the activity of mitochondrial respiration.

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β-Lapachone Induces Acute Oxidative Stress in Rat Primary Astrocyte Cultures that is Terminated by the NQO1-Inhibitor Dicoumarol

Cited by 11 publications

References 60 publications

β-lapachone-mediated WST1 Reduction as Indicator for the Cytosolic Redox Metabolism of Cultured Primary Astrocytes

β-lapachone-mediated WST1 Reduction as Indicator for the Cytosolic Redox Metabolism of Cultured Primary Astrocytes

Tumor Microenvironment Triggered Cascade‐Activation Nanoplatform for Synergistic and Precise Treatment of Hepatocellular Carcinoma

Consumption and Metabolism of Extracellular Pyruvate by Cultured Rat Brain Astrocytes

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