Functional Characterization of Ergothioneine Transport by Rat Organic Cation/Carnitine Transporter Octn1 (slc22a4)

Nakamura, Toshimichi; Yoshida, Kenji; Yabuuchi, Hidetake; Maeda, Tomoji; Tamai, Ikumi

doi:10.1248/bpb.31.1580

Cited by 49 publications

(36 citation statements)

References 23 publications

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“…The fact that nonradiolabeled ergothioneine could compete with [ 3 H]ergothioneine in these experiments strongly indicates that uptake occurs via a specific transporter rather than by passive diffusion. OCTN-1 transports ergothioneine more efficiently when sodium is present (Gründemann et al, 2005;Nakamura et al, 2008). This earlier finding was corroborated by the present study, which demonstrated that [ 3 H]ergothioneine uptake in endothelial cells is also sodiumdependent.…”

Section: The [supporting

confidence: 90%

“…Furthermore, transfection of the OCTN-1 gene into human embryonic kidney 293 cells allows uptake of ergothioneine and reduces Cu 21 -induced cytotoxicity (Nakamura et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

“…As a consequence, ergothioneine is a stable antioxidant that is nonautoxidizable at physiologic pH (Hartman, 1990). Ergothioneine is positively charged at physiologic pH, and requires the specific carrier protein organic cation transporter novel type-1 (OCTN-1) to cross the cell membrane (Grigat et al, 2007;Nakamura et al, 2008). OCTN-1 is mainly found in kidney, trachea, and bone marrow (Tamai et al, 1997;Koepsell et al, 2007), but its existence in other cell types has not been explored.…”

Section: Introductionmentioning

confidence: 99%

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Uptake and Protective Effects of Ergothioneine in Human Endothelial Cells

Yang²,

Sit

et al. 2014

J Pharmacol Exp Ther

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Ergothioneine is a thiourea derivative of histidine found in food, especially mushrooms. Experiments in cell-free systems and chemical assays identified this compound as a powerful antioxidant. Experiments were designed to test the ability of endothelial cells to take up ergothioneine and hence benefit from protection against oxidative stress. Reverse-transcription polymerase chain reaction and Western blotting demonstrated transcription and translation of an ergothioneine transporter in human brain microvascular endothelial cells (HBMECs). Uptake of [3 H]ergothioneine occurred by the organic cation transporter novel type-1 (OCTN-1), was sodium-dependent, and was reduced when expression of OCTN-1 was silenced by small interfering RNA (siRNA). The effect of ergothioneine on the production of reactive oxygen species (ROS) in HBMECs was measured using dichlorodihydrofluorescein and lucigenin, and the effect on cell viability was studied using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. ROS production and cell death induced by pyrogallol, xanthine oxidase plus xanthine, and high glucose were suppressed by ergothioneine. The antioxidant and cytoprotective effects of ergothioneine were abolished when OCTN-1 was silenced using siRNA. The expression of NADPH oxidase 1 was decreased, and those of glutathione reductase, catalase, and superoxide dismutase enhanced by the compound. In isolated rat basilar arteries, ergothioneine attenuated the reduction in acetylcholine-induced relaxation caused by pyrogallol, xanthine oxidase plus xanthine, or incubation in high glucose. Chronic treatment with the compound improved the response to acetylcholine in arteries of rats with streptozotocin-induced diabetes. In summary, ergothioneine is taken up by endothelial cells via OCTN-1, where the compound then protects against oxidative stress, curtailing endothelial dysfunction.

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Section: The [supporting

confidence: 90%

“…Furthermore, transfection of the OCTN-1 gene into human embryonic kidney 293 cells allows uptake of ergothioneine and reduces Cu 21 -induced cytotoxicity (Nakamura et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Uptake and Protective Effects of Ergothioneine in Human Endothelial Cells

Yang²,

Sit

et al. 2014

J Pharmacol Exp Ther

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“…When mouse OCTN1 or OCTN2 was heterologously transfected in HEK293 cells, only OCTN1, but not OCTN2 efficiently transports ergothioneine as a substrate (Figure 2A), as previously reported for human and rat OCTN1 (3,19). The uptake of [ 3 H]ergothioneine by mouse OCTN1 was saturable with Km and Vmax values of 4.68 µM and 532 pmol/min/mg protein, respectively ( Figure 2B) and reduced by the replacement of Na + in the medium ( Figure 2C).…”

Section: Ergothioneine Is a Substrate Of Mouse Octn1supporting

confidence: 84%

Gene Knockout and Metabolome Analysis of Carnitine/Organic Cation Transporter OCTN1

et al. 2010

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Purpose: Solute carrier, OCTN1 (SLC22A4) is an orphan transporter, the physiologically important substrate of which is still unidentified. The aim of the present study was to examine physiological roles of OCTN1. Methods:We first constructed octn1 gene knockout (octn1 -/-) mice. Metabolome analysis was then performed to identify substrates in vivo. The possible association of the substrate identified with diseased conditions was further examined. Results:The metabolome analysis of blood and several organs indicated complete deficiency of a naturally occurring potent antioxidant ergothioneine in octn1 -/-mice among 112 metabolites examined.Pharmacokinetic analyses after oral administration revealed the highest distribution to small intestines and extensive renal reabsorption of [ 3 H]ergothioneine, both of which were much reduced in octn1 -/-mice.The octn1 -/-mice exhibited greater susceptibility to intestinal inflammation under the ischemia and reperfusion model. The blood ergothioneine concentration was also much reduced in Japanese patients with Crohn's disease, compared with healthy volunteers and patients with another inflammatory bowel disease, ulcerative colitis. 4Conclusions: These results indicate that OCTN1 plays a pivotal role for maintenance of systemic and intestinal exposure of ergothioneine which could be important for protective effect against intestinal tissue injuries, providing a possible diagnostic tool to distinguish the inflammatory bowel diseases.

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“…HEK293 cells overexpressing rat and human organic cation transporter isoforms HEK/rOctn2, HEK/hOCTN1, and HEK/hOCTN2 were previously established in our laboratory (Tamai et al, 2001;Fujita et al, 2009), whereas HEK/rOctn1 was transfected for this study using methods similar to those described previously (Nakamura et al, 2008). HEK293 cells were cultured using a 10-cm 2 culture dish in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 g/ml streptomycin, 6 mg/ml HEPES, and 1 mM pyruvic acid under a humidified atmosphere at 37°C and 5% CO 2 -95% air.…”

Section: Methodsmentioning

confidence: 99%

Oxaliplatin Transport Mediated by Organic Cation/Carnitine Transporters OCTN1 and OCTN2 in Overexpressing Human Embryonic Kidney 293 Cells and Rat Dorsal Root Ganglion Neurons

Jong¹,

Nakanishi²,

Liu³

et al. 2011

J Pharmacol Exp Ther

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112

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The organic cation/carnitine transporters OCTN1 and OCTN2 are related to other organic cation transporters (OCT1, OCT2, and OCT3) known for transporting oxaliplatin, an anticancer drug with dose-limiting neurotoxicity. In this study, we sought to determine whether OCTN1 and OCTN2 also transported oxaliplatin and to characterize their functional expression and contributions to its neuronal accumulation and neurotoxicity in dorsal root ganglion (DRG) neurons relative to those of OCTs. [ 14 C]Oxaliplatin uptake, platinum accumulation, and cytotoxicity were determined in OCTN-overexpressing human embryonic kidney (HEK) 293 cells and primary cultures of rat DRG neurons. Levels of mRNA and functional activities of rat (r)Octns and rOcts in rat DRG tissue and primary cultures were characterized using reverse transcription-polymerase chain reaction and uptake of model OCT/OCTN substrates, including [ showed increased uptake and cytotoxicity of oxaliplatin compared with mock-transfected HEK293 controls; in addition, both uptake and cytotoxicity were inhibited by ergothioneine and L-carnitine. The uptake of ergothioneine mediated by OCTN1 and of L-carnitine mediated by OCTN2 was decreased during oxaliplatin exposure. rOctn1 and rOctn2 mRNA was readily detected in rat DRG tissue, and they were functionally active in cultured rat DRG neurons, more so than rOct1, rOct2, or rOct3. DRG neuronal accumulation of [ 14 C]oxaliplatin and platinum during oxaliplatin exposure depended on time, concentration, temperature, and sodium and was inhibited by ergothioneine and to a lesser extent by L-carnitine but not by MPP ϩ . Loss of DRG neuronal viability during oxaliplatin exposure was inhibited by ergothioneine but not by L-carnitine or MPP ϩ . OCTN1 and OCTN2 both transport oxaliplatin and are functionally expressed by DRG neurons. OCTN1-mediated transport of oxaliplatin appears to contribute to its neuronal accumulation and treatmentlimiting neurotoxicity more so than OCTN2 or OCTs.

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Functional Characterization of Ergothioneine Transport by Rat Organic Cation/Carnitine Transporter Octn1 (slc22a4)

Cited by 49 publications

References 23 publications

Uptake and Protective Effects of Ergothioneine in Human Endothelial Cells

Uptake and Protective Effects of Ergothioneine in Human Endothelial Cells

Gene Knockout and Metabolome Analysis of Carnitine/Organic Cation Transporter OCTN1

Oxaliplatin Transport Mediated by Organic Cation/Carnitine Transporters OCTN1 and OCTN2 in Overexpressing Human Embryonic Kidney 293 Cells and Rat Dorsal Root Ganglion Neurons

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