Continuous exposure to l-arginine induces oxidative stress and physiological tolerance in cultured human endothelial cells

Mohan, Srinidi; Wu, C. S. Wilson; Shin, Soyoung; Fung, Ho‐Leung

doi:10.1007/s00726-011-1173-y

Cited by 36 publications

(35 citation statements)

References 35 publications

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“…In whole cells and isolated cell membranes, ADMA increased cellular DHE fluorescence at concentrations above 10 μM, indicating that intracellular proteins are not critical for this action. ADMA-induced DHE fluorescence was inhibited by L-lysine and L-NAME, suggesting that cellular uptake and interaction with eNOS were necessary, consistent with the mechanisms of oxidative stress induced by ARG that we have shown recently [25]. Addition of BH 4 abrogated ADMA-induced cellular oxidative stress and reduced NO availability, indicating that the oxidative effects of ADMA may be mediated via eNOS uncoupling.…”

Section: Discussionsupporting

confidence: 87%

Mechanism of Cellular Oxidation Stress Induced by Asymmetric Dimethylarginine

Mohan

Fung

2012

IJMS

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The mechanism by which asymmetric dimethylarginine (ADMA) induces vascular oxidative stress is not well understood. In this study, we utilized human umbilical vein endothelial cells (HUVEC) to examine the roles of ADMA cellular transport and the uncoupling of endothelial nitric oxide synthase (eNOS) in contributing to this phenomenon. Dihydroethidium (DHE) fluorescence was used as an index of oxidative stress. Whole cells and their isolated membrane fractions exhibited measureable increased DHE fluorescence at ADMA concentrations greater than 10 μM. ADMA-induced DHE fluorescence was inhibited by co-incubation with L-lysine, tetrahydrobiopterin (BH4), or L-nitroarginine methyl ester (L-NAME). Oxidative stress induced in these cells by angiotensin II (Ang II) were unaffected by the same concentrations of L-lysine, L-NAME and BH4. ADMA-induced reduction in cellular nitrite or nitrite/nitrate production was reversed in the presence of increasing concentrations of BH4. These results suggest that ADMA-induced DHE fluorescence involves the participation of both the cationic transport system in the cellular membrane and eNOS instead of the Ang II-NADPH oxidase pathway.

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Section: Discussionsupporting

confidence: 87%

Mechanism of Cellular Oxidation Stress Induced by Asymmetric Dimethylarginine

Mohan

Fung

2012

IJMS

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“…We cannot exclude the possibility that intracellular specific L-arginine pool is depleted because of up-regulation of Arg-II. However, this concept is not supported by studies showing that continuous cellular exposure to L-arginine for 7 days does not affect L-arginine uptake and generation of the endogenous eNOS inhibitor ADMA [46]. Our previous study showed that Arg-II causes eNOS-uncoupling requiring L-arginine metabolizing enzymatic activity [25].…”

Section: Discussionmentioning

confidence: 99%

Long term exposure to L-arginine accelerates endothelial cell senescence through arginase-II and S6K1 signaling

Xiong

Fru

et al. 2014

Aging

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L-arginine supplementation is proposed to improve health status or as adjunct therapy for diseases including cardiovascular diseases. However, controversial results and even detrimental effects of L-arginine supplementation are reported. We investigate potential mechanisms of L-arginine-induced detrimental effects on vascular endothelial cells. Human endothelial cells were exposed to a physiological (0.1 mmol/L) or pharmacological (0.5 mmol/L) concentration of L-arginine for 30 minutes (acute) or 7 days (chronic). The effects of L-arginine supplementation on endothelial senescence phenotype, i.e., levels of senescence-associated beta-galactosidase, expression of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1, eNOS-uncoupling, arginase-II expression/activity, and mTORC1-S6K1 activity were analyzed. While acute L-arginine treatment enhances endothelial NO production accompanied with superoxide production and activation of S6K1 but no up-regulation of arginase-II, chronic L-arginine supplementation causes endothelial senescence, up-regulation of the adhesion molecule expression, and eNOS-uncoupling (decreased NO and enhanced superoxide production), which are associated with S6K1 activation and up-regulation of arginase-II. Silencing either S6K1 or arginase-II inhibits up-regulation/activation of each other, prevents endothelial dysfunction, adhesion molecule expression, and senescence under the chronic L-arginine supplementation condition. These results demonstrate that S6K1 and arginase-II form a positive circuit mediating the detrimental effects of chronic L-arginine supplementation on endothelial cells.

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“…Since concurrent work showed that high concentrations of glucose (4.5 G/L) and ARG (84 mg/L) in the original DMEM medium inhibited eNOS activity [23], a further-modified DMEM, which contained 0.9 G/L of glucose and 21 mg/L of ARG was used for ADMA metabolism studies. Cells were used for experiments after they reached confluence in 6-well plates in 7 days.…”

Section: Methodsmentioning

confidence: 99%

Cellular interactions between L-arginine and asymmetric dimethylarginine: Transport and metabolism

2017

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This study was aimed to examine the effect of L-arginine (ARG) exposure on the disposition of asymmetric dimethylarginine (ADMA) in human endothelial cells. Although the role of ADMA as an inhibitor of endothelial nitric oxide synthase (eNOS) is well-recognized, cellular interactions between ARG and ADMA are not well-characterized. EA.hy926 human vascular endothelial cells were exposed to 15N4-ARG, and the concentrations of 15N4-ARG and ADMA in the cell lysate and incubation medium were determined by a liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) assay. Nitric oxide (NO) production was estimated by utilizing cumulative nitrite concentration via a fluorometric assay. Cells incubated with 15N4-ARG exhibited enhanced nitrite production as well as 15N4-ARG cellular uptake. These changes were accompanied by a decrease in cellular ADMA level and increase in extracellular ADMA level, indicating an efflux of endogenous ADMA from the cell. The time courses of ADMA efflux as well as nitrite accumulation in parallel with 15N4-ARG uptake were characterized. Following preincubation with 15N4-ARG and D7-ADMA, the efflux of cellular 15N4-ARG and D7-ADMA was significantly stimulated by high concentrations of ARG or ADMA in the incubation medium, demonstrating trans-stimulated cellular transport of these two amino acids. D7-ADMA metabolism was inhibited in the presence of added ARG. These results demonstrated that in addition to an interaction at the level of eNOS, ARG and ADMA may mutually influence their cellular availability via transport and metabolic interactions.

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Continuous exposure to l-arginine induces oxidative stress and physiological tolerance in cultured human endothelial cells

Cited by 36 publications

References 35 publications

Mechanism of Cellular Oxidation Stress Induced by Asymmetric Dimethylarginine

Mechanism of Cellular Oxidation Stress Induced by Asymmetric Dimethylarginine

Long term exposure to L-arginine accelerates endothelial cell senescence through arginase-II and S6K1 signaling

Cellular interactions between L-arginine and asymmetric dimethylarginine: Transport and metabolism

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