metallothionein and epigallocatechin-gallate

Catechins and their polymers procyanidins are health-promoting flavonoids found in edible vegetables and fruits. They act as antioxidants by scavenging reactive oxygen species and by chelating the redox-active metals iron and copper. They also behave as signaling molecules, modulating multiple cell signalling pathways and gene expression, including that of antioxidant enzymes. This study aimed at determining whether catechins and procyanidins interact with the redox-inactive metal zinc and at assessing their effect on cellular zinc homeostasis. We found that a grape-seed procyanidin extract (GSPE) and the green tea flavonoid (-)-epigallocatechin-3-gallate (EGCG) bind zinc cations in solution with higher affinity than the zinc-specific chelator Zinquin, and dose-dependently prevent zinc-induced toxicity in the human hepatocarcinoma cell line HepG2, evaluated by the lactate dehydrogenase test. GSPE and EGCG hinder intracellular accumulation of total zinc, measured by atomic flame absorption spectrometry, concomitantly increasing the level of cytoplasmic labile zinc detectable by Zinquin fluorescence. Concurrently, GSPE and EGCG inhibit the expression, evaluated at the mRNA level by quantitative reverse transcriptase-polymerase chain reaction, of zinc-binding metallothioneins and of plasma membrane zinc exporter ZnT1 (SLC30A1), while enhancing the expression of cellular zinc importers ZIP1 (SLC39A1) and ZIP4 (SLC39A4). GSPE and EGCG also produce all these effects when HepG2 cells are stimulated to import zinc by treatment with supplemental zinc or the proinflammatory cytokine interleukin-6. We suggest that extracellular complexation of zinc cations and the elevation of cytoplasmic labile zinc may be relevant mechanisms underlying the modulation of diverse cell signaling and metabolic pathways by catechins and procyanidins.

Topics: Biflavonoids; Carcinoma, Hepatocellular; Catechin; Cation Transport Proteins; Diet; Gene Expression Regulation; Grape Seed Extract; Hep G2 Cells; Homeostasis; Humans; Interleukin-6; Liver Neoplasms; Metallothionein; Proanthocyanidins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Tea; Zinc

The tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) has been studied for chronic disease preventive effects, and is marketed as part of many dietary supplements. However, case-reports have associated the use of green tea-based supplements with liver toxicity. We studied the hepatotoxic effects of high dose EGCG in male CF-1 mice. A single dose of EGCG (1500 mg/kg, i.g.) increased plasma alanine aminotransferase (ALT) by 138-fold and reduced survival by 85%. Once-daily dosing with EGCG increased hepatotoxic response. Plasma ALT levels were increased 184-fold following two once-daily doses of 750 mg/kg, i.g. EGCG. Moderate to severe hepatic necrosis was observed following treatment with EGCG. EGCG hepatotoxicity was associated with oxidative stress including increased hepatic lipid peroxidation (5-fold increase), plasma 8-isoprostane (9.5-fold increase) and increased hepatic metallothionein and gamma-histone 2AX protein expression. EGCG also increased plasma interleukin-6 and monocyte chemoattractant protein-1. Our results indicate that higher bolus doses of EGCG are hepatotoxic to mice. Further studies on the dose-dependent hepatotoxic effects of EGCG and the underlying mechanisms are important given the increasing use of green tea dietary supplements, which may deliver much higher plasma and tissue concentrations of EGCG than tea beverages.

Topics: Aldehydes; Animals; Antioxidants; Biomarkers; Catechin; Chemical and Drug Induced Liver Injury; Chromatography, High Pressure Liquid; Cysteine; Cytokines; Dose-Response Relationship, Drug; Immunohistochemistry; Lipid Peroxidation; Liver Function Tests; Male; Metallothionein; Mice; Oxidants; Oxidative Stress; Spectrometry, Mass, Electrospray Ionization; Survival Analysis

This study was conducted to explore the effects of epigallocatechin-3-gallate (EGCG) on cognitive performances in psychological stress rats. An animal model of psychological stress was developed by restraint stress for three weeks. Male Wistar rats were randomly assigned to four groups as follows: normal control group, stress control group and two stress groups with green tea polyphenols (GTPs) and EGCG modulation, respectively. The changes of behavioral performances of rats were examined by the open-field test and step-through test. Results showed that behavioral performances of stress control group were changed abnormally, and they were improved in GTPs and EGCG modulation groups. In addition, plasma levels of cortisol, dopamine, norepinephrine, 5-hydroxytryptamine, interleukin-6 and interleukin-2 were detected. Stress control group had increased contents of cortisol, interleukin-6 and interleukin-2, and meanwhile had declined levels of 5-hydroxytryptamine and catecholamines. These changes in GTPs and EGCG modulation groups were similar to that of the normal control group. The expressions of metallothioneins in the hippocampus were detected by reverse transcription polymerase chain reaction. In contrast with the normal control group, their expressions in all the three stress groups were enhanced clearly. The results suggested that GTPs and EGCG modulation could improve the cognitive impairments induced by psychological stress. The related mechanisms may be involved with the changes of catecholamines, 5-hydroxytryptamine, cytokines and expressions of metallothioneins.

Topics: Animals; Behavior, Animal; Catechin; Catecholamines; Gene Expression Regulation; Hippocampus; Hydrocortisone; Interleukin-2; Interleukin-6; Male; Metallothionein; Phytotherapy; Rats; Rats, Wistar; Serotonin; Stress, Psychological