Distribution and accumulation of dietary ergothioneine and its metabolites in mouse tissues

Richard Ming Yi Tang, Irwin Kee-Mun Cheah, Terry Shze Keong Yew, Barry Halliwell, Richard Ming Yi Tang, Irwin Kee-Mun Cheah, Terry Shze Keong Yew, Barry Halliwell

Abstract

L-ergothioneine (ET) is a diet-derived amino acid that accumulates at high concentrations in animals and humans. Numerous studies have highlighted its antioxidant abilities in vitro, and possible cytoprotective capabilities in vivo. We investigated the uptake and distribution of ET in various organs by a highly sensitive and specific liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) technique, both before and after oral administration of pure ET (35 and 70 mg/kg/day for 1, 7, and 28 days) to male C57BL6J mice. ET primarily concentrates in the liver and whole blood, and also in spleen, kidney, lung, heart, intestines, eye, and brain tissues. Strong correlations were found between ET and its putative metabolites - hercynine, ET-sulfonate (ET-SO3H), and S-methyl ET. Hercynine accumulates in the brain after prolonged ET administration. This study demonstrates the uptake and distribution of ET and provides a foundation for future studies with ET to target oxidative damage in a range of tissues in human diseases.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Structure of ergothioneine (ET) and its possible metabolites. (a) ET exists predominantly in the thione form at physiological pH. Putative metabolic derivatives of ET – (b) hercynine, (c) ET sulfonate (ET-SO3H), and (d) S-methyl ET.
Figure 2
Figure 2
Experimental setup. C57BL6-J mice (n = 7) were orally gavaged with 35 mg/kg (ET+) or 70 mg/kg (ET++) of ET every day for 1, 7 and 28 days. Control mice were gavaged with saline. Mice were euthanized 24 h after the last administration, and organs were harvested.
Figure 3
Figure 3
Distribution and accumulation of ET in mouse liver, whole blood, spleen, kidney, lung, heart, small intestine (jejunum/ileum), eye, large intestine, and brain (cortex). (a) Basal ET concentration in mice administered with single dose of saline (1-day control), n = 7. (b) Maximum ET accumulation in mice fed with 70 mg/kg/day ET for 28 days, n = 7. (c) Rate of ET accumulation was determined by calculating the fold change between maximum and basal ET concentrations. All values are expressed with standard error.
Figure 4
Figure 4
Uptake and accumulation of ET in various mouse tissues. (a–j) Mice (n = 7) were orally administered with either saline, 35 mg/kg (ET+) or 70 mg/kg (ET++) of ET per day for 1, 7 or 28 days, and 10 tissues (liver, whole blood, spleen, kidney, lung, heart, small intestine (jejunum/ileum), eye, large intestine, and brain) were harvested and ET concentration measured by LC-MS/MS. Spleen, kidney, and lung data were corrected for blood ET contribution. (k) RT-PCR of OCTN1 mRNA expression in liver of mice fed with saline (Control) or 70 mg/kg/day ET (ET++) for 1 and 28 days (n = 7); Two-tailed Mann-Whitney test **p < 0.01. All values are expressed with standard error.
Figure 5
Figure 5
Basal tissue distribution of ET related metabolites in mice fed with single dose of saline (1-day control), n = 7. (a) Hercynine and ratio of ET to hercynine concentration, (b) ET-SO3H concentrations, and (c) S-methyl ET concentrations. All values are expressed with standard error.
Figure 6
Figure 6
Accumulation of hercynine in mouse tissues. (a–j) As per ET concentration quantified in Fig. 4, hercynine levels were determined simultaneously by LC-MS/MS in liver, whole blood, spleen, kidney, lung, heart, small intestine (jejunum/ileum), eye, large intestine, and brain. Spleen, kidney, and lung hercynine data were corrected for blood hercynine contribution. All values are expressed with standard error.
Figure 7
Figure 7
Accumulation of ergothioneine sulfonate (ET-SO3H) in mouse tissues. (a–g) Similar to hercynine, ET-SO3H levels were quantified by LC-MS/MS in whole blood, spleen, kidney, lung, heart, small intestine (jejunum/ileum), and large intestine. All values are expressed with standard error.
Figure 8
Figure 8
Accumulation of S-methyl ET in mouse tissues. (a–h) Similar to hercynine, S-methyl ET levels were quantified by LC-MS/MS in whole blood, spleen, kidney, lung, heart, small intestine (jejunum/ileum), and large intestine. All values are expressed with standard error.
Figure 9
Figure 9
Correlation of ET concentration with hercynine, ET-SO3H, and S-methyl ET. Other correlations can be found in Supplementary data 3. (a–d) Correlation of whole blood, kidney, heart and lung ET to hercynine. (e–g) Correlation of liver, eye and lung ET to S-methyl ET. (h–i) Correlation of whole blood and kidney ET to ET-SO3H. All values are expressed with standard error.

References

    1. Tanret, C. Sur une base nouvelle retiree du seigle ergote, l’ergothioneine. Compt Rend, 222–224 (1909).
    1. Carlsson J, Kierstan MP, Brocklehurst K. Reactions of L-ergothioneine and some other aminothioneswith2,2′-and 4,4′-dipyridyl disulphides and of L-ergothioneine with iodoacetamide. 2-Mercaptoimidazoles, 2- and 4-thiopyridones, thiourea and thioacetamide as highly reactive neutral sulphur nucleophils. The Biochemical journal. 1974;139:221–235. doi: 10.1042/bj1390221.
    1. Bello MH, Barrera-Perez V, Morin D, Epstein L. The Neurospora crassa mutant NcDeltaEgt-1 identifies an ergothioneine biosynthetic gene and demonstrates that ergothioneine enhances conidial survival and protects against peroxide toxicity during conidial germination. Fungal Genet Biol. 2012;49:160–172. doi: 10.1016/j.fgb.2011.12.007.
    1. Sheridan KJ, et al. Ergothioneine Biosynthesis and Functionality in the Opportunistic Fungal Pathogen, Aspergillus fumigatus. Sci Rep. 2016;6:35306. doi: 10.1038/srep35306.
    1. Pluskal T, Ueno M, Yanagida M. Genetic and metabolomic dissection of the ergothioneine and selenoneine biosynthetic pathway in the fission yeast, S. pombe, and construction of an overproduction system. PLoS One. 2014;9:e97774. doi: 10.1371/journal.pone.0097774.
    1. Genghof DS, Inamine E, Kovalenko V, Melville DB. Ergothioneine in microorganisms. The Journal of biological chemistry. 1956;223:9–17.
    1. Genghof DS, Vandamme O. Biosynthesis of ergothioneine and hercynine by mycobacteria. Journal of bacteriology. 1964;87:852–862.
    1. Narainsamy K, et al. Oxidative-stress detoxification and signalling in cyanobacteria: the crucial glutathione synthesis pathway supports the production of ergothioneine and ophthalmate. Mol Microbiol. 2016;100:15–24. doi: 10.1111/mmi.13296.
    1. Pfeiffer C, Bauer T, Surek B, Schömig E, Gründemann D. Cyanobacteria produce high levels of ergothioneine. Food Chemistry. 2011;129:1766–1769. doi: 10.1016/j.foodchem.2011.06.047.
    1. Seebeck FP. In vitro reconstitution of Mycobacterial ergothioneine biosynthesis. J Am Chem Soc. 2010;132:6632–6633. doi: 10.1021/ja101721e.
    1. Alamgir KM, Masuda S, Fujitani Y, Fukuda F, Tani A. Production of ergothioneine by Methylobacterium species. Front Microbiol. 2015;6:1185. doi: 10.3389/fmicb.2015.01185.
    1. Cheah IK, Halliwell B. Ergothioneine; antioxidant potential, physiological function and role in disease. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease. 2012;1822:784–793. doi: 10.1016/j.bbadis.2011.09.017.
    1. Cheah IK, Tang RM, Yew TS, Lim KH, Halliwell B. Administration of Pure Ergothioneine to Healthy Human Subjects: Uptake, Metabolism, and Effects on Biomarkers of Oxidative Damage and Inflammation. Antioxid Redox Signal. 2017;26:193–206. doi: 10.1089/ars.2016.6778.
    1. Grundemann D, et al. Discovery of the ergothioneine transporter. Proceedings of the National Academy of Sciences of the United States of America. 2005;102:5256–5261. doi: 10.1073/pnas.0408624102.
    1. Kato Y, et al. Gene knockout and metabolome analysis of carnitine/organic cation transporter OCTN1. Pharmaceutical research. 2010;27:832–840. doi: 10.1007/s11095-010-0076-z.
    1. Ey J, Schomig E, Taubert D. Dietary sources and antioxidant effects of ergothioneine. Journal of agricultural and food chemistry. 2007;55:6466–6474. doi: 10.1021/jf071328f.
    1. Weigand-Heller AJ, Kris-Etherton PM, Beelman RB. The bioavailability of ergothioneine from mushrooms (Agaricus bisporus) and the acute effects on antioxidant capacity and biomarkers of inflammation. Prev Med. 2012;54:Suppl, S75–78. doi: 10.1016/j.ypmed.2011.12.028.
    1. Kalaras MD, Richie JP, Calcagnotto A, Beelman RB. Mushrooms: A rich source of the antioxidants ergothioneine and glutathione. Food Chem. 2017;233:429–433. doi: 10.1016/j.foodchem.2017.04.109.
    1. Melville DB, Horner WH, Lubschez R. Tissue ergothioneine. The Journal of biological chemistry. 1954;206:221–228.
    1. Shires TK, Brummel MC, Pulido JS, Stegink LD. Ergothioneine distribution in bovine and porcine ocular tissues. Comparative biochemistry and physiology Part C, Pharmacology, toxicology & endocrinology. 1997;117:117–120. doi: 10.1016/S0742-8413(96)00223-X.
    1. Salt HB. The ergothioneine content of the blood in health and disease. The Biochemical journal. 1931;25:1712–1719. doi: 10.1042/bj0251712.
    1. Leone E, Mann T. Ergothioneine in the seminal vesicle secretion. Nature. 1951;168:205–206. doi: 10.1038/168205b0.
    1. Halliwell B, Cheah IK, Drum CL. Ergothioneine, an adaptive antioxidant for the protection of injured tissues? A hypothesis. Biochem Biophys Res Commun. 2016;470:245–250. doi: 10.1016/j.bbrc.2015.12.124.
    1. Franzoni F, et al. An in vitro study on the free radical scavenging capacity of ergothioneine: comparison with reduced glutathione, uric acid and trolox. Biomedecine & pharmacotherapie. 2006;60:453–457. doi: 10.1016/j.biopha.2006.07.015.
    1. Rougee M, Bensasson RV, Land EJ, Pariente R. Deactivation of singlet molecular oxygen by thiols and related compounds, possible protectors against skin photosensitivity. Photochemistry and photobiology. 1988;47:485–489. doi: 10.1111/j.1751-1097.1988.tb08835.x.
    1. Colognato R, et al. Modulation of hydrogen peroxide-induced DNA damage, MAPKs activation and cell death in PC12 by ergothioneine. Clin Nutr. 2006;25:135–145. doi: 10.1016/j.clnu.2005.10.005.
    1. Laurenza I, Colognato R, Migliore L, Del Prato S, Benzi L. Modulation of palmitic acid-induced cell death by ergothioneine: evidence of an anti-inflammatory action. Biofactors. 2008;33:237–247. doi: 10.1002/biof.5520330401.
    1. Rahman I, et al. Ergothioneine inhibits oxidative stress- and TNF-alpha-induced NF-kappa B activation and interleukin-8 release in alveolar epithelial cells. Biochem Biophys Res Commun. 2003;302:860–864. doi: 10.1016/S0006-291X(03)00224-9.
    1. Motohashi N, Mori I, Sugiura Y, Tanaka H. Radioprotective effect of ergothioneine on gamma-irradiation of metmyoglobin: comparison with cysteine on sulfmyoglobin-formation. Chem Pharm Bull (Tokyo) 1977;25:2516–2523. doi: 10.1248/cpb.25.2516.
    1. Hartman PE, Hartman Z, Citardi MJ. Ergothioneine, histidine, and two naturally occurring histidine dipeptides as radioprotectors against gamma-irradiation inactivation of bacteriophages T4 and P22. Radiat Res. 1988;114:319–330. doi: 10.2307/3577228.
    1. Hseu YC, et al. Dermato-protective properties of ergothioneine through induction of Nrf2/ARE-mediated antioxidant genes in UVA-irradiated Human keratinocytes. Free Radic Biol Med. 2015;86:102–117. doi: 10.1016/j.freeradbiomed.2015.05.026.
    1. Cheah IK, et al. Liver ergothioneine accumulation in a guinea pig model of non-alcoholic fatty liver disease. A possible mechanism of defence? Free Radic Res. 2016;50:14–25. doi: 10.3109/10715762.2015.1099642.
    1. Moncaster JA, Walsh DT, Gentleman SM, Jen LS, Aruoma OI. Ergothioneine treatment protects neurons against N-methyl-D-aspartate excitotoxicity in an in vivo rat retinal model. Neurosci Lett. 2002;328:55–59. doi: 10.1016/S0304-3940(02)00427-5.
    1. Song TY, Chen CL, Liao JW, Ou HC, Tsai MS. Ergothioneine protects against neuronal injury induced by cisplatin both in vitro and in vivo. Food Chem Toxicol. 2010;48:3492–3499. doi: 10.1016/j.fct.2010.09.030.
    1. Cheah IK, Feng L, Tang RM, Lim KH, Halliwell B. Ergothioneine levels in an elderly population decrease with age and incidence of cognitive decline; a risk factor for neurodegeneration? Biochem Biophys Res Commun. 2016;478:162–167. doi: 10.1016/j.bbrc.2016.07.074.
    1. Hatano T, Saiki S, Okuzumi A, Mohney RP, Hattori N. Identification of novel biomarkers for Parkinson’s disease by metabolomic technologies. Journal of neurology, neurosurgery, and psychiatry. 2016;87:295–301. doi: 10.1136/jnnp-2014-309676.
    1. Servillo L, D’Onofrio N, Balestrieri ML. Ergothioneine Antioxidant Function: From Chemistry to Cardiovascular Therapeutic Potential. J Cardiovasc Pharmacol. 2017;69:183–191. doi: 10.1097/FJC.0000000000000464.
    1. Marone PA, Trampota J, Weisman S. A Safety Evaluation of a Nature-Identical l-Ergothioneine in Sprague Dawley Rats. Int J Toxicol. 2016;35:568–583. doi: 10.1177/1091581816653375.
    1. Nakamichi N, et al. Food-derived hydrophilic antioxidant ergothioneine is distributed to the brain and exerts antidepressant effect in mice. Brain Behav. 2016;6:e00477. doi: 10.1002/brb3.477.
    1. Servillo L, et al. An uncommon redox behavior sheds light on the cellular antioxidant properties of ergothioneine. Free Radic Biol Med. 2015;79:228–236. doi: 10.1016/j.freeradbiomed.2014.11.017.
    1. Sotgia S, et al. Identification of the Main Intermediate Precursor of l-Ergothioneine Biosynthesis in Human Biological Specimens. Molecules. 2016;21:1298. doi: 10.3390/molecules21101298.
    1. Bachmanov AA, Reed DR, Beauchamp GK, Tordoff MG. Food intake, water intake, and drinking spout side preference of 28 mouse strains. Behav Genet. 2002;32:435–443. doi: 10.1023/A:1020884312053.
    1. Mitsuyama H, May JM. Uptake and antioxidant effects of ergothioneine in human erythrocytes. Clin Sci (Lond) 1999;97:407–411. doi: 10.1042/cs0970407.
    1. Wang LZ, et al. Quantification of L-ergothioneine in human plasma and erythrocytes by liquid chromatography-tandem mass spectrometry. J Mass Spectrom. 2013;48:406–412. doi: 10.1002/jms.3150.
    1. Arduini A, Eddy L, Hochstein P. The reduction of ferryl myoglobin by ergothioneine: a novel function for ergothioneine. Arch Biochem Biophys. 1990;281:41–43. doi: 10.1016/0003-9861(90)90410-Z.
    1. Bedirli A, et al. Ergothioneine pretreatment protects the liver from ischemia-reperfusion injury caused by increasing hepatic heat shock protein 70. J Surg Res. 2004;122:96–102. doi: 10.1016/j.jss.2004.06.016.
    1. Spicer SS, Wooley JG, Kessler V. Ergothioneine depletion in rabbit erythrocytes and its effect on methemoglobin formation and reversion. Proc Soc Exp Biol Med. 1951;77:418–420. doi: 10.3181/00379727-77-18799.
    1. Kawano H, Higuchi F, Mayumi T, Hama T. Studies on ergothioneine. VII. Some effects on ergothioneine on glycolytic metabolism in red blood cells from rats. Chem Pharm Bull (Tokyo) 1982;30:2611–2613. doi: 10.1248/cpb.30.2611.
    1. Shinozaki Y. et al. Impairment of the carnitine/organic cation transporter 1-ergothioneine axis is mediated by intestinal transporter dysfunction in chronic kidney disease. Kidney Int, In press (2017).
    1. Santosa S, Jones PJ. Oxidative stress in ocular disease: does lutein play a protective role? CMAJ. 2005;173:861–862. doi: 10.1503/cmaj.1031425.
    1. Kimura A, et al. Targeting Oxidative Stress for Treatment of Glaucoma and Optic Neuritis. Oxid Med Cell Longev. 2017;2017:2817252. doi: 10.1155/2017/2817252.
    1. Abu-Amero KK, Kondkar AA, Chalam KV. Resveratrol and Ophthalmic Diseases. Nutrients. 2016;8:200. doi: 10.3390/nu8040200.
    1. Kim EC, et al. Screening and Characterization of Drugs That Protect Corneal Endothelial Cells Against Unfolded Protein Response and Oxidative Stress. Invest Ophthalmol Vis Sci. 2017;58:892–900. doi: 10.1167/iovs.16-20147.
    1. Wu X, et al. Structural and functional characteristics and tissue distribution pattern of rat OCTN1, an organic cation transporter, cloned from placenta. Biochim Biophys Acta. 2000;1466:315–327. doi: 10.1016/S0005-2736(00)00189-9.
    1. Nakamura T, Yoshida K, Yabuuchi H, Maeda T, Tamai I. Functional characterization of ergothioneine transport by rat organic cation/carnitine transporter Octn1 (slc22a4) Biol Pharm Bull. 2008;31:1580–1584. doi: 10.1248/bpb.31.1580.
    1. Jang JH, Aruoma OI, Jen LS, Chung HY, Surh YJ. Ergothioneine rescues PC12 cells from beta-amyloid-induced apoptotic death. Free Radic Biol Med. 2004;36:288–299. doi: 10.1016/j.freeradbiomed.2003.11.005.
    1. Ishimoto T, et al. Organic cation transporter-mediated ergothioneine uptake in mouse neural progenitor cells suppresses proliferation and promotes differentiation into neurons. PLoS One. 2014;9:e89434. doi: 10.1371/journal.pone.0089434.
    1. Heath H, Rimington C, Searle CE, Lawson A. Some effects of administering ergothioneine to rats. The Biochemical journal. 1952;50:530–533. doi: 10.1042/bj0500530.
    1. Heath H. The metabolism of 35S-labelled 2-thiolhistidine and ergothioneine in the rat. The Biochemical journal. 1953;54:689–694. doi: 10.1042/bj0540689.
    1. Grigat S, et al. Probing the substrate specificity of the ergothioneine transporter with methimazole, hercynine, and organic cations. Biochem Pharmacol. 2007;74:309–316. doi: 10.1016/j.bcp.2007.04.015.
    1. Askari A, Melville DB. The reaction sequence in ergothioneine biosynthesis: hercynine as an intermediate. The Journal of biological chemistry. 1962;237:1615–1618.
    1. Redford-Ellis M, Gowenlock AH. Studies on the reaction of chloromethane with human blood. Acta Pharmacol Toxicol (Copenh) 1971;30:36–48. doi: 10.1111/j.1600-0773.1971.tb00632.x.
    1. Sinclair, P. R., Gorman, N. & Jacobs, J. M. Measurement of heme concentration. Curr Protoc Toxicol Chapter 8, Unit8 3 (2001).

Source: PubMed

Patrocinadores e Colaboradores

Condições médicas

Intervenções de drogas

3
Se inscrever