Concentrations Of Ergothioneine Research Articles

Ergothioneine, an adaptive antioxidant for the protection of injured tissues? A hypothesis

Ergothioneine (ET) is a diet-derived, thiolated derivative of histidine with antioxidant properties. Although ET is produced only by certain fungi and bacteria, it can be found at high concentrations in certain human and animal tissues and is absorbed through a specific, high affinity transporter (OCTN1). In liver, heart, joint and intestinal injury, elevated ET concentrations have been observed in injured tissues. The physiological role of ET remains unclear. We thus review current literature to generate a specific hypothesis: that the accumulation of ET in vivo is an adaptive mechanism, involving the regulated uptake and concentration of an exogenous natural compound to minimize oxidative damage.

Organic cation transporter Octn1-mediated uptake of food-derived antioxidant ergothioneine into infiltrating macrophages during intestinal inflammation in mice

OCTN1/SLC22A4 is expressed on apical membranes of small intestine, and is involved in gastrointestinal absorption of its substrates, including the food-derived antioxidant ergothioneine (ERGO). ERGO concentration in circulating blood of patients with inflammatory bowel disease (Crohn's disease) is lower than that in healthy volunteers; thus, circulating ERGO is a potential diagnostic marker, although the mechanisms underlying low ERGO concentration in patients are unknown. Here, we focused on intestinal macrophages, which infiltrate sites of inflammation, and examined possible first-pass uptake of ERGO by macrophages. ERGO concentration in blood was lower in mice with dextran sodium sulfate (DSS)-induced colitis than in controls. On the other hand, expression of octn1 gene product and ERGO concentration in intestinal tissues of DSS-treated mice were higher than in controls. Interestingly, lamina propria mononuclear cells (LPMCs) isolated from DSS-treated mice contained ERGO and showed [3H]ERGO uptake and Octn1 expression, whereas ERGO was undetectable in LPMCs of control mice. Functional expression of OCTN1 was also confirmed in LPS-stimulated human macrophage-like cell line, THP-1. In conclusion, OCTN1 is functionally expressed on activated intestinal macrophages, and ERGO uptake into these immune cells could contribute at least in part to the altered disposition of ERGO in intestinal inflammation.

Endogenous ergothioneine is required for wild type levels of conidiogenesis and conidial survival but does not protect against 254 nm UV-induced mutagenesis or kill

Ergothioneine, a histidine derivative, is concentrated in conidia of ascomycetous fungi. To investigate the function of ergothioneine, we crossed the wild type Neurospora crassa (Egt+) and an ergothioneine non-producer (Egt−, Δegt-1, a knockout in NCU04343.5) and used the Egt+ and Egt− progeny strains for phenotypic analyses. Compared to the Egt+ strains, Egt− strains had a 59% reduction in the number of conidia produced on Vogel’s agar. After storage of Egt+ and Egt− conidia at 97% and 52% relative humidity (RH) for a time course to either 17 or 98days, respectively, Egt− strains had a 23% and a 18% reduction in life expectancy at 97% and 52% RH, respectively, compared to the Egt+ strains. Based on a Cu(II) reduction assay with the chelator bathocuproinedisulfonic acid disodium salt, ergothioneine accounts for 38% and 33% of water-soluble antioxidant capacity in N. crassa conidia from seven and 20day-old cultures, respectively. In contrast, ergothioneine did not account for significant (α=0.05) anti-oxidant capacity in mycelia, which have lower concentrations of ergothioneine than conidia. The data are consistent with the hypothesis that ergothioneine has an antioxidant function in vivo. In contrast, experiments on the spontaneous mutation rate in Egt+ and Egt− strains and on the effects of 254nm UV light on mutation rate and conidial viability do not support the hypothesis that ergothioneine protects DNA in vivo.

Optimization of process variables for supercritical fluid extraction of ergothioneine and polyphenols from Pleurotus ostreatus and correlation to free-radical scavenging activity

Supercritical carbon dioxide extraction was employed to extract antioxidants from Pleurotus ostreatus. The response surface methodology was employed to determine the optimal conditions for extraction of ergothioneine and polyphenols. The ergothioneine concentration in the mushroom extract was quantified and characterized using high pressure liquid chromatography (HPLC) followed by tandem mass spectrometry (MS/MS). The optimized values of responses were obtained at a pressure of 21MPa, a temperature of 48°C and a co-solvent amount of 133ml, yielding an ergothioneine content of 1.35mg/g dw, total phenol content of 5.48mgGAE/g dw, and IC50 for DPPH radical scavenging capacity of 0.008mg/ml. A higher desirability value of 0.98 for model demonstrated that response surface methodology can be successfully applied for optimizing supercritical carbon dioxide extraction of antioxidants from P. ostreatus. A good correlation was found between DPPH radical scavenging capacity and ergothioneine (R2=0.94) as well as with polyphenols (R2=0.95).

Effect of mushroom diet on pharmacokinetics of gabapentin in healthy Chinese subjects

This study evaluated the pharmacokinetics of gabapentin in Chinese subjects who received a diet rich in shiitake mushrooms. Shiitake mushrooms have been shown to contain high amount of ergothioneine. In vitro studies have shown that OCTN1-mediated secretion of gabapentin is trans-stimulated by ergothioneine. This study also investigated the concentrations of ergothioneine in plasma at baseline and following mushroom consumption. Ten healthy male subjects were recruited and received a diet containing no mushrooms (treatment A) or a high mushroom diet (treatment B; after at least a 7 day washout period) 1 day prior to administration of a single oral dose of gabapentin 600 mg. Ingestion of shiitake mushrooms produced significant increases in plasma ergothioneine concentrations that were sustained for more than 48 h. A statistically significant but modest increase in the renal clearance (CLR ) of gabapentin occurred after intake of the mushroom diet (91.1 ± 25.1 vs. 76.9 ± 20.6 ml min(-1) , P = 0.031). No significant changes in AUC(0,tlast ) of gabapentin were observed (P = 0.726). Creatinine clearance did not correlate with CLR of gabapentin at baseline (treatment A). After ingestion of the mushroom diet, creatinine clearance accounted for 65.3% of the variance in CLR of gabapentin. These data suggest that diet-drug pharmacokinetic interactions may occur during co-exposure to gabapentin and mushroom constituents. However, as it does not affect the AUC(0,tlast ) of gabapentin, it may not have clinically important consequences. Shiitake mushrooms can also be used as a source of ergothioneine for future clinical studies.

Effects of UV-B Radiation Levels on Concentrations of Phytosterols, Ergothioneine, and Polyphenolic Compounds in Mushroom Powders Used As Dietary Supplements

Compositional changes of powder dietary supplements made from mushrooms exposed to different levels of UV-B irradiation were evaluated for the bioactive naturally occurring mushroom antioxidant, ergothioneine; other natural polyphenolic compounds, e.g., flavonoids, lignans, etc.; and selected phytosterols. Four types of mushroom powder consisting of white, brown (Agaricus bisporus), oyster (Pleurotus ostreatus), and shiitake (Lentinula edodes) mushrooms from three different treatment groups (control, low and high UV-B exposures) were evaluated. Ergothioneine concentrations found in mushroom powders were 0.4-10.4 mg/g dry weight (dw) and were not appreciably affected by UV-B radiation. No individual polyphenols were detected above 0.1 μg/g. Phytosterols ergosterol (2.4-6.2 mg/g dw) and campesterol (14-43 μg/g dw) were measured in mushroom powder samples. Ergosterol concentrations decreased significantly with the increased level of UV-B treatment for all mushroom powder types, except for white. These results provide some new information on effects of UV-B radiation on these important natural bioactive compounds in mushrooms.

재배조건과 수확시기에 따른 천마의 기능성 물질함량 변화

Gastrodia elata Blume is a achlorophyllous orchid plant that has been used in traditional medicine to treat many human illnesses, such as vertigo, hemiplegia, and epilepsy. It contains three major pharmacological substances; gastrodin, ergothioneine, and vanillyl alcohol. Here we investigated the quantitative changes in those functional components by cultivation conditions and harvest times. The content of ergothioneine was different with cultivation areas, while gastrodin accumulated the higher levels than vanillyl alcohol regardless of cultivation areas. The smaller tubers contained the higher concentrations of gastrodin and the lower concentrations of ergothioneine than did the larger tubers. The cultivation under sun shade screen increased the concentrations of both gastrodin and vanillyl alcohol but decreased the levels of ergothioneine. We also determined the contents of functional substances depending on the harvest times. The concentration of gastrodin was higher in October than in April, while the level of ergothioneine was higher in April than in October. Interestingly, the use of sexually propagated seed tubers produced 1.9 times higher levels of gastrodin compared to vegetatively propagated seed tubers. In this study, we found that three major functional substances in G. elata tubers altered depending on various cultivation practices, which might be important to increase the economic values of G. elata tubers.

Ergothioneine attenuates the DNA damage of post-thawed Merino ram sperm

The objective of the current study was to evaluate the effects of antioxidant ergothioneine added to cryopreservation extender on the DNA integrity of Merino ram sperm. Semen samples from 5 mature Merino rams (1 and 2years of age) were used in the study. Semen samples, which were diluted with a Tris-based extender containing ergothioneine at different concentrations and no antioxidant (control), were cooled to 5°C and frozen in 0.25ml French straws. Frozen straws were then thawed at 37°C for 20s in a water bath for evaluation of sperm DNA damage using the Comet test. The addition of ergothioneine at concentrations of 1, 2 and 4mM resulted in lower sperm with damaged DNA (5.4, 4.7 and 3.2%, respectively) than that of control (7.9%), thus reducing the DNA damage (P<0.01). Findings of this study showed that the increasing concentrations of ergothioneine in semen extenders, were of greater benefit to DNA integrity of frozen-thawed ram sperm.

The Effect of Ergothioneine on Clastogenic Potential and Mutagenic Activity

L-(+)-ergothioneine has antioxidant and anti-inflammatory properties in vitro and in vivo and has uses as a dietary supplement and as an ingredient in foods, cosmetics, and as a pharmaceutical additive. The clastogenic potential and mutagenic of ergothioneine were assessed in vitro and in vivo. Ergothioneine concentrations up to 5000 μg/mL, with and without metabolic activation, was tested in the chromosome aberration assay with CHL cells and found not to induce structural chromosome aberrations. In the in vivo mammalian erythrocyte micronucleus test, ergothioneine was administered orally to male mice at doses up to 1500 mg/kg for potential genotoxic activity. No increase in the frequency of micronucleated polychromatic erythrocytes was observed. Overall, ergothioneine was not genotoxic in these studies and provides additional experimental evidence supporting the safety of its use as a potential dietary supplement.

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

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. 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. 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. These results indicate that OCTN1 plays a pivotal role for maintenance of systemic and intestinal exposure of ergothioneine, which could be important for protective effects against intestinal tissue injuries, providing a possible diagnostic tool to distinguish the inflammatory bowel diseases.

Imidazole-Based Erythrocyte Markers of Oxidative Stress in Preeclampsia—An NMR Investigation

Using (1)H-nuclear magnetic resonance (NMR) spectroscopy and statistical models, we sought to identify ''biomarkers'' present in erythrocytes that would distinguish between women with normal pregnancy and those suffering from preeclampsia, and investigate possible links with previously identified plasma ''markers.'' Erythrocytes from 22 normotensive pregnant women and 15 preeclamptics were analyzed by (1)H Carr-Purcell-Meiboom-Gill (CPMG) NMR. Multivariate analysis and logistic regression were applied to differentiate between the 2 groups of patients, and used to develop a diagnostic model based on the concentrations of the constituents identified as being influential. Significantly higher concentrations of alanine (P < .001), glycine (P = .025), and ergothioneine (P = .049) were found in erythrocytes from preeclamptic patients. Discriminant analysis and regression of NMR data permitted 100% accurate diagnosis of the health status of new patients. Chemically related imidazole-based molecules, histidine and ergothioneine, are important in the classification process and the etiology of preeclampsia (PE).

Enhancement of the antioxidants ergothioneine and selenium in Pleurotus eryngii var. eryngii basidiomata through cultural practices

Antioxidants are molecules that may reverse, prevent or slow cellular damage caused by free radicals. Increasing dietary intake of antioxidants is thought to reduce oxidative stress that may contribute to the development of several diseases. Mushrooms are known to contain antioxidants such as selenium, ergothioneine and phenolics that may serve this role. Here we sought to enhance selenium and ergothioneine concentration in Pleurotus eryngii var. eryngii basidiomata by modifying the techniques used for their commercial cultivation. To enhance selenium content in mushrooms, substrates were supplemented with sodium selenite (Na2SeO3) to reach selenium concentrations of 5 and 10 μg/g. Basidiomata of one commercial isolate (WC888) accumulated selenium up to 4.6 and 9.3 μg/g (d.w.), respectively. Therefore, a serving size (85 g) of fresh P. eryngii mushrooms produced on substrates supplemented with 5 and 10 μg/g of Na2SeO3 would supply 70.4 and 116.3% of the daily value of selenium (DV = 70 μg), respectively. Since selenium-enriched mushrooms would supply more than 20% of the DV, they could be considered an excellent source of selenium. Ergothioneine concentration was enhanced in mushrooms produced on low-moisture (55%) substrate compared to the commonly used 60% (high-moisture) in commercial cultivation. Mushrooms produced on low-moisture substrate had ergothioneine concentrations of 3.0 mg/g, while mushrooms produced on high-moisture substrate contained 2.2 mg/g or less. Use of a casing overlay for mushroom production resulted in significant yield increases on low-moisture substrate but not on high-moisture substrate.

Uptake and antioxidant effects of ergothioneine in human erythrocytes

Ergothioneine is a fungal metabolite that may have antioxidant functions in mammalian cells. Although it accumulates to low millimolar concentrations in liver and other tissues, it is not thought to be taken up by mature erythrocytes. During a study of the function of ergothioneine as an antioxidant in human erythrocytes, we found that these cells do take up ergothioneine from the surrounding medium. Ergothioneine concentrations in freshly prepared erythrocytes were 2-9-fold higher than in plasma from the same donor. Slow but progressive accumulation of ergothioneine to about 125% of basal levels was observed in erythrocytes over a 4 h incubation. After a 2 h incubation, intracellular ergothioneine concentrations rose on addition of increasing amounts of ergothioneine to the incubation medium, although saturation was not evident in cells from all donors. Both initial levels and rates of ergothioneine uptake varied in erythrocytes from different donors. Intracellular ergothioneine was stable to depletion of GSH by N-ethylmaleimide and to a more severe oxidant stress induced by hydrogen peroxide in the presence of catalase. These results show that human erythrocytes do take up ergothioneine; however, the GSH results do not support an antioxidant role for ergothioneine in erythrocytes.

Influence of age and season on certain biochemical constituents of seminal plasma of Arabian horses

The study used 36 mature (4–25 years of age) and healthy Arabian horses reared at a government stud near Cairo. A total of 75 ejaculates was collected from these animals during a complete annual cycle and included 53 ejaculates from 30 stallions to estimate the influence of consecutive seasons and ages on seminal plasma concentrations of glycerylphosphorylcholine (GPC), ergothioneine (EGT), fructose, citric acid, total protein (PRT) and total carbohydrate. The respective overall mean (±SEM) values of these constituents were 75.91 ± 3.44 mg per 100 ml, 13.86 ± 0.79 mg per 100 ml, 7.85 ± 0.56 mg per 100 ml, 44.12 ± 1.83 mg per 100 ml, 644.92 ± 31.72 mg per 100 ml and 39.74 ± 1.57 mg per 100 ml seminal plasma. There were statistically significant age-related effects on seminal plasma concentrations of GPC, EGT and PRT, with seasonal effects on all six measured constituents. Modulation of the selected seminal plasma constituents by age and season must be considered when selecting Arabian stallions for fertility.

Comparison of the 1H and 31P NMR Spectra of Erythrocytes and Plasma from some Australian Native Animals: Bandicoot, Echidna, Koala, Little Penguin, Tammar Wallaby, Tasmanian Devil, Tree Kangaroo and Wombat

1H and 31P NMR spectra were acquired from the whole plasma and ultrafiltrates of erythrocytes (RBC) from eight Australian native animals. Various metabolites contained in the samples were identified from their NMR spectra and their concentrations were determined where possible. The key observations were as follows. (1) High lysine concentrations in the RBC from Tammar wallaby (∼8 mmol/(L RBC)) were confirmed. (2) A resonance at δ 3.17 in the 1H spin-echo NMR spectrum of koala RBC was reassigned to the N+-(CH3)3 of betaine and not to ergothioneine as previously claimed. (3) A high concentration (∼10 mmol/ (L RBC)) of taurine was observed in the RBC from ‘little’ penguin. (4) The lack of ATP in the RBC of echidna, reported previously on the basis of standard biochemical assays, was confirmed directly by NMR spectroscopy. (5) The highest ergothioneine concentrations in the RBC of the eight species of animal were in the wombat (1.76 mmol/(L RBC)).

Study of the ergothioneine concentration in the blood of individuals with diabetes mellitus.

Ergothioneine concentrations in the blood of 113 patients with diabetes mellitus and 22 non-diabetic individuals were measured by high performance liquid chromatography. There is no statistically significant difference between the mean ergothioneine concentrations of the diabetic and non-diabetic populations but there are several significant differences within the diabetic population. Male diabetics have a higher ergothioneine concentration than female diabetics. Type II diabetics have a higher ergothioneine concentration than type I diabetics. Diabetics receiving a higher insulin dose have higher ergothioneine concentrations. There was no correlation of ergothioneine concentration with the concentration of haemoglobin A1 or with the number of years diabetic. It is suggested that zinc therapy may be beneficial for diabetics having a high concentration of blood ergothioneine.

Interception of some direct-acting mutagens by ergothioneine.

Ergothioneine, a novel imidazole sulfhydryl/thione compound formed in millimolar amounts by fungi, is a potentially important defense against electrophiles and free radicals. Protection may well occur both in organisms that synthesize ergothioneine and in animals including man that ingest and store ergothioneine in red blood cells, the liver, seminal fluid, and central nervous system. Ergothioneine blocks the mutagenicity for Salmonella strain TA1950 (hisG46 uvrB) of the nitrosation products of spermidine to an extent that is approximately proportional to the ergothioneine concentration. Ergothioneine also alleviates mutagenicity of cumene and t-butyl hydroperoxides but does not react with N-methyl-N'nitro-N-nitrosoguanidine as does the cysteinyl sulfhydryl compound, glutathione.

The role of ergothioneine in the oxidation of reduced nicotinamide adenine dinucleotide by metmyoglobin or methemoglobin.

In phosphate buffer (pH 6.8), reduced nicotinamide adenine dinucleotide (NADH) was oxidized by metmyoglobin or methemoglobin in the presence of Mn (II) ion and ergothioneine. The NADH oxidation was dependent on the concentrations of Mn (II) ion, ergothioneine and metmyoglobin (or methemoglobin). Various phosphorus compounds also influenced the oxidation. Orthophosphate was the most effective at concentrations or more than 50 mM. In contrast, nucleotides such as adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP) and adenosine-5'-monophosphate (AMP) were less effective. Superoxide dismutase or catalase inhibited the present NADH oxidation, suggesting the participation of superoxide anion (O-2) and H2O2 in this metmyoglobin system as well as the peroxidase system. Metmyoglobin pretreated with H2O2, i.e., ferrylmyoglobin, promoted the NADH oxidation reaction, but oxymyoglobin inhibited it. A mechanism is proposed for the oxidation of NADH by metmyoglobin or methemoglobin on the basis of the redox potential of ergothioneine, the formation of Mn-phosphate complex and the heme state of metmyoglobin or methemoglobin.

Biochemical components of stallion seminal plasma before and after the breeding season

Seasonal and breed differences were determined for glyceryl-phosphoryl-choline (GPC), ergothioneine (EGT), fructose and total protein in the seminal plasma of 24 stallions from three breed groups. Significant differences were found between the GPC, EGT, fructose and total protein levels before and after the breeding season. During the post-breeding period, the concentrations of EGT, fructose and total protein increased more than 100% while that for GPC decreased by about 50%. This phenomenon was found in each of the breed groups investigated. Breed differences in the GPC level during the pre- and post-seasonal periods were not significant. Differences were found only during the pre-seasonal period in EGT, fructose and total protein levels.