gluconasturtiin and phenethyl-isothiocyanate

Cruciferous vegetables are rich sources of glucosinolates which are the biogenic precursor molecules of isothiocyanates (ITCs). The relationship between the consumption of cruciferous vegetables and chemoprotection has been widely documented in epidemiological studies. Phenethyl isothiocyanate (PEITC) occurs as its glucosinolate precursor gluconasturtiin in the cruciferous vegetable watercress (Nasturtium officinale). PEITC has multiple biological effects, including activation of cytoprotective pathways, such as those mediated by the transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) and the transcription factor heat shock factor 1 (HSF1), and can cause changes in the epigenome. However, at high concentrations, PEITC leads to accumulation of reactive oxygen species and cytoskeletal changes, resulting in cytotoxicity. Underlying these activities is the sulfhydryl reactivity of PEITC with cysteine residues in its protein targets. This chemical reactivity highlights the critical importance of the dose of PEITC for achieving on-target selectivity, which should be carefully considered in the design of future clinical trials.

Topics: Animals; Cell Line, Tumor; Cytoprotection; Epigenesis, Genetic; Gene Expression Regulation; Glucosinolates; Heat Shock Transcription Factors; Humans; Isothiocyanates; Nasturtium; NF-E2 Transcription Factor, p45 Subunit; NF-E2-Related Factor 2; Reactive Oxygen Species; Vegetables

Horseradish (Armoracia rusticana) is a perennial crop and its ground root tissue is used in condiments because of the pungency of the glucosinolate (GS)-hydrolysis products allyl isothiocyanate (AITC) and phenethyl isothiocyanate (PEITC) derived from sinigrin and gluconasturtiin, respectively. Horseradish roots are sold in three grades: U.S. Fancy, U.S. No. 1, and U.S. No. 2 according to the USDA standards. These grading standards are primarily based on root diameter and length. There is little information on whether root grades vary in their phytochemical content or potential health promoting properties. This study measured GS, GS-hydrolysis products, potential anticancer activity (as quinone reductase inducing activity), total phenolic content, and antioxidant activities from different grades of horseradish accessions. U.S. Fancy showed significantly higher sinigrin and AITC concentrations than U.S. No. 1 ,whereas U.S. No. 1 showed significantly higher concentrations of 1-cyano 2,3-epithiopropane, the epithionitrile hydrolysis product of sinigrin, and significantly higher total phenolic concentrations than U.S. Fancy.

Topics: Armoracia; Genotype; Glucosinolates; Isothiocyanates; Plant Proteins; Plant Roots; Quinone Reductases

In this study, the effect of myrosinase-treated glucoerucin (GER+MYR), which releases the isothiocyanate (ITC) erucin, on heme oxygenase 1 (HO-1) gene expression and Nrf2 signaling was investigated in vitro in cultured cells and in vivo in mice. Treatment of HT-29 cells with GER+MYR resulted in a significant increase in the mRNA and protein levels of nuclear Nrf2 and HO-1. GER+MYR was more potent at enhancing the nuclear Nrf2 levels than were the following myrosinase-treated glucosinolates: sinigrin, glucoraphanin and gluconasturtiin, which are the precursors of allyl-ITC, R-sulforaphane and 2-phenylethyl ITC, respectively. GER+MYR also significantly induced HO-1 gene expression in the mouse intestinal mucosae and liver but not in the brain. Mechanistic studies suggest that GER+MYR induces Nrf2 via ERK1/2-, p38- and JNK-dependent signal transduction pathways. The GER+MYR-mediated increase in HO-1 expression is primarily attributable to p38 signaling.

Topics: Animals; Brain; Diet, High-Fat; Female; Glucose; Glucosinolates; Glycoside Hydrolases; Heme Oxygenase-1; HT29 Cells; Humans; Imidoesters; Intestinal Mucosa; Intestines; Isothiocyanates; Liver; Membrane Proteins; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinase 3; Mustard Plant; NF-E2-Related Factor 2; Oximes; p38 Mitogen-Activated Protein Kinases; Plant Extracts; RNA, Messenger; Signal Transduction; Sulfoxides; Up-Regulation

Methyl jasmonate spray treatments (250 μM) were utilized to alter glucosinolate composition in the florets of the commercial broccoli F1 hybrids 'Pirate', 'Expo', 'Green Magic', 'Imperial', and 'Gypsy' grown in replicated field plantings in 2009 and 2010. MeJA treatment significantly increased glucoraphanin (11%), gluconasturtiin (59%), and neoglucobrassicin (248%) concentrations and their hydrolysis products including sulforaphane (152%), phenethyl isothiocyanate (318%), N-methoxyindole-3-carbinol (313%), and neoascorbigen (232%) extracted from florets of these genotypes over two seasons. Increased quinone reductase (QR) activity was significantly correlated with increased levels of sulforaphane, N-methoxyindole-3-carbinol, and neoascorbigen. Partitioning experiment-wide trait variances indicated that the variability in concentrations of sulforaphane (29%), neoascorbigen (48%), and QR activity (72%) was influenced by year-associated weather variables, whereas variation in neoglucobrassicin (63%) and N-methoxyindole-3-carbinol (46%) concentrations was primarily attributed to methyl jasmonate treatment. These results suggest that methyl jasmonate treatment can enhance QR inducing activity by increased hydrolysis of glucoraphanin into sulforaphane and the hydrolysis products of neoglucobrassicin.

Topics: Acetates; Brassica; Cyclopentanes; Glucosinolates; Hydrolysis; Imidoesters; Indoles; Isothiocyanates; NAD(P)H Dehydrogenase (Quinone); Oximes; Oxylipins; Plant Proteins; Seasons; Sulfoxides

Glucosinolates are compounds produced by commercial crops which can hydrolyse in a range of natural toxins that may exert detrimental effects on beneficial soil organisms. This study examined the effects of 2-phenylethyl isothiocyanate and 3-phenylpropionitrile on the survival and growth of the woodlouse Porcellio scaber exposed for 28 d. 2-Phenylethyl isothiocyanate dissipated from the soil with half-lives ranging from 19 to 96 h. Exposure through soil showed toxic effects only on survival. The LC50s after 28 d were significantly different at 65.3 mg kg(-1) for 2-phenylethyl isothiocyanate and 155 mg kg(-1) for 3-phenylpropionitrile. A toxicokinetic-toxicodynamic (TKTD) approach, however, revealed that both compounds in fact have very similar effect patterns. The TKTD model was better suited to interpret the survival data than descriptive dose-response analysis (LC(x)), accounting for the fast dissipation of the compounds in the soil. Found effects were within environmentally relevant concentrations. Care should therefore be taken before allowing these natural toxins to enter soil ecosystems in large quantities.

Topics: Animals; Glucosinolates; Hydrolysis; Isopoda; Isothiocyanates; Soil; Soil Pollutants

Cruciferous vegetable consumption is associated with decreased risk of several cancers, including prostate cancer. Gluconasturtiin, one of the predominant glucosinolates in cruciferous vegetables, is hydrolyzed to yield phenylethyl isothiocyanate (PEITC). PEITC absorption and metabolism in humans involves glutathione conjugation followed by conversion via the mercapturic acid pathway to an N-acetylcysteine (NAC) conjugate that is excreted in the urine. We observed an inhibitory effect of PEITC and its metabolite, NAC-PEITC, on cancer cell proliferation, cell-cycle progression, and apoptosis in LNCaP human prostate cancer cells. PEITC and NAC-PEITC suppressed LNCaP cell proliferation in a dose-dependent manner, and exposure to 5 microM PEITC or NAC-PEITC reduced cell proliferation by 25% and 30%, respectively. Cell-cycle analysis revealed that cells treated with 5 microM PEITC or NAC-PEITC arrested at the G(2)/M phase. In addition, the percentage of cells in the S phase decreased from 46% to 25% following 48 h of incubation with PEITC or NAC-PEITC. The G(2)/M-phase cell-cycle arrest of LNCaP cells grown in the presence of PEITC or NAC-PEITC is correlated with the downregulation of Cdk1 and cyclin B(1) protein expression. Apoptosis was observed at the later stages of 24-h and 48-h treatments with 5 microM PEITC and NAC-PEITC. In conclusion, PEITC and NAC-PEITC are potential chemopreventive/chemotherapeutic agents against LNCaP human prostate cancer cells.

Topics: Acetylcysteine; Antineoplastic Agents, Phytogenic; Apoptosis; Brassicaceae; CDC2 Protein Kinase; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cyclin B1; Dose-Response Relationship, Drug; Down-Regulation; Gene Expression; Glucosinolates; Humans; Isothiocyanates; Male; Phytotherapy; Plant Extracts; Prostatic Neoplasms

The effect of gluconasturtiin (GNST) and phenethyl isothiocyanate (PEITC) on some metabolic changes and antioxidative parameters in the rat was tested using different doses of PEITC and duration of GNST or PEITC ingestion. Their effect on antioxidative processes was previously observed, however, their influence on metabolic changes is still poorly characterized. In the performed experiment, the effect of GNST (0.5 mg/kg BW) and PEITC (0.1 mg/kg BW or 0.3 mg/kg BW) administered intragastrically after 4 h or 14 days to growing male rats was studied. PEITC at both doses after 4 h of its administration caused a considerable increase in liver cholesterol and triglyceride content with a concomitant drop in the amount of glycogen. Blood glucose, free fatty acids, phospholipids and total, free, esterified cholesterol as well as cholesterol in high-density lipoprotein were not altered. GNST, at its short-time ingestion, augmented significantly the concentration of triglycerides in blood serum. The compounds tested had no influence on metabolic changes after a longer period of action with the exception of glycogen values in liver, which were substantially augmented by PEITC at both doses. Our trial revealed a lack of GNST and PEITC influence on the content of liver sulphhydryl groups and on glutathione peroxidase and glutathione-S-transferase activities. The only distinct change in the content of malonodialdehyde was observed after short-time action of lower dose of PEITC. Our research showed that the short-term PEITC action constituted a significant factor interfering with liver metabolism. Although PEITC has been repeatedly advocated as very promising anticancer agent, in our experiment, the lower dose of PEITC was revealed as a pro-oxidative substance. These inconsistent properties seem to depend on its dose and time of action.

Topics: Animals; Anticarcinogenic Agents; Antioxidants; Blood Glucose; Cholesterol; Dose-Response Relationship, Drug; Energy Metabolism; Glucosinolates; Isothiocyanates; Liver; Liver Glycogen; Male; Malondialdehyde; Oxidation-Reduction; Rats; Rats, Wistar; Triglycerides

It is thought that induction of detoxifying phase-II drug metabolizing enzymes or inhibition of bioactivating phase-I by phytoalexins could protect against mutagens and neoplasia. In the search for potential naturally occurring molecular chemoprevention agents, particular attention has been devoted to isothiocyanates, which are breakdown products-via myrosinase-of glucosinolates such as gluconasturtiin (GNST), a natural constituent of cruciferae. Here, we first investigated the ability of GNST to modulate metabolizing enzymes in male Swiss Albino CD1 mice injected by gavage (24 mg/kg or 48 mg/kg b.w.) with GNST either in single or repeated (daily for four consecutive days) dose. Using selected probes to various cytochrome P450 (CYP) isoforms, a marked and generalized decrease of CYP content, NADPH-(CYP)-c-reductase and various CYP-linked monooxygenases (measuring CYP1A1, CYP2B1/2, CYP3A1/2, CYP1A2 and CYP2E1), was observed in hepatic, renal and pulmonary subcellular preparations (up to approximately 66% loss, liver). Similar behavior was recorded using the regio- and stereo-selective hydroxylation of testosterone as multibiomarker (CYP2A1 and CYP2B9, up to approximately 96% loss), as well as with the phase-II marker glutathione S-transferase (up to approximately 50% loss, liver). We also performed genotoxicity investigations, using the diploid D7 strain of yeast Saccharomyces cerevisiae as a biological test system. GNST was able to significantly induce point reverse mutation in growing cells without myrosinase, thus suggesting either a direct GNST or a CYP-linked metabolite role in the genotoxic response. On the contrary, in suspension test, the addition of myrosinase significantly increased mitotic gene conversion, probably due to the formation of GNST-derived phenylethyl isothiocyanate (PEITC) breakdown product. Taken together, our data suggest that GNST exerts a dual effect: while strongly inhibiting the microsomal (bioactivating) metabolism, GNST also possesses genotoxic activity. This concomitant mutagenic activity underlines the necessity of overall toxicological characterization of this (or any other molecule) prior to mass chemopreventive use.

Topics: Animals; Brassicaceae; Cytochrome P-450 Enzyme System; Enzyme Inhibitors; Glucosinolates; Isothiocyanates; Kidney; Liver; Lung; Male; Mice; Mutagens; Saccharomyces cerevisiae; Xenobiotics

Cytotoxic and cell-transforming activity of gluconasturtiin (GNST), a promising chemopreventive agent commonly found in human diet, was studied in a medium-term bioassay utilizing BALB/c 3T3 cells. We also assessed whether GNST coupled with myrosinase, thus yielding product phenylethyl isothiocyanate (as shown by gas chromatography-mass spectral analysis), can affect the transforming potential of benzo(a)pyrene (B(a)P). Neither cytotoxicity nor cell-transforming activity was recorded. On the contrary, a marked increase (up to sevenfold) of the transforming activity of B(a)P was seen. This cocarcinogenic potential could be ascribed to an imbalance among bioactivation/detoxication during cell growth. These results indicate the need for an overall toxicological characterization of a chemopreventive agent prior to large-scale use.

Topics: 3T3 Cells; Aldehyde Dehydrogenase; Animals; Benzo(a)pyrene; Brassica; Carcinogens; Cell Survival; Cell Transformation, Neoplastic; Cocarcinogenesis; Drug Synergism; Glucosinolates; Glycoside Hydrolases; In Vitro Techniques; Isothiocyanates; Mice; Mice, Inbred BALB C; Risk Factors

The isolation of gram-amounts of 2-phenylethyl glucosinolate (gluconasturtiin, GST) from Barbarea verna seeds is reported for the first time. This vegetable source was of crucial importance to isolate GST with a high purity grade and in high yield. Indeed, B. verna seeds contain GST as the only glucosinolate, unlike other sources. The availability at low cost of GST will allow further studies to explain the claimed anticancer activity of its derived phenylethyl isothiocyanate.

Topics: Aldehyde Dehydrogenase; Anticarcinogenic Agents; Brassica; Chromatography, High Pressure Liquid; Glucosinolates; Humans; Isothiocyanates; Phytotherapy; Plant Extracts; Seeds