Toxicogenomics of resveratrol in rat liver
Introduction
Resveratrol (trans-3,4′,5-trihydroxystilbene) is a polyphenolic compound found in plant sources, such as the skin of grapes (Jang et al., 1997, Soleas et al., 1997) and peanuts (Sanders et al., 2000). Resveratrol is attributed with varied biological properties including anti-oxidant and anti-inflammatory effects. These properties were observed from the beneficial effects of red wine in preventing cardiovascular disease, which contains resveratrol (1.5–3 mg/L of red wine); the phenomenon is referred to as “French Paradox” due to the consumption of cheese and other high fat foods in the daily diet and the relatively low incidence of heart disease in that geographical region (Goldberg, 1996). In addition, resveratrol was found to interfere with cellular events leading to tumor initiation, promotion and progression (Jang et al., 1997). It blocked the formation of preneoplastic lesions in mouse mammary glands (Jang et al., 1997) and has been shown to inhibit proliferation of a variety of cancer cells in culture including human colon cancer cells, breast epithelial cells and prostate cancer cells (Mgbonyebi et al., 1998, Mitchell et al., 1999, Schneider et al., 2000).
Several mechanisms have been proposed for resveratrol's anti-cancer and chemopreventive action. Resveratrol's ability to inhibit the enzyme cyclooxygenase (COX) has been proposed as one of the plausible mechanisms of action (Shin et al., 1998, Subbaramaiah et al., 1998). Resveratrol inhibits COX-1 non-competitively in a dose-dependent manner and modulates COX-2 by inhibiting the PKC signal transduction pathway. Polyphenols obtained from foods, which include resveratrol inhibit pancreatic cancer growth and cause apoptosis by the release of cytochrome c and activation of caspases (Mouria et al., 2002). Resveratrol's action on inhibiting the proliferation of cancer cells is accompanied by cell cycle arrest in a large number of cancer cell lines (Ahmad et al., 2001, Bernhard et al., 2000, Joe et al., 2002, Schneider et al., 2000).
Resveratrol is also known to have an effect on the cell's drug metabolizing enzyme systems, such as, Phase I and Phase II drug metabolizing enzymes (DME's). In particular, resveratrol was found to inhibit cytochrome P450 1A1 (CYP1A1), 1A2 (CYP1A2) and 2E1 (CYP2E1) drug metabolizing enzymes (Chang et al., 2001, Chun et al., 1999, Ciolino and Yeh, 1999, Mikstacka et al., 2002). Furthermore, another important effect of resveratrol is the induction of the Phase II enzyme NAD(P)H:quinone oxidoreductase in cultured mouse hepatoma (Hepa1c1c7) cells (Prochaska and Santamaria, 1988). The induction of Phase II enzymes plays an important role in detoxifying harmful substances. Indeed, the induction of these enzymes is an essential factor to consider when studying chemopreventive properties of a compound (Gerhauser et al., 1997, Hecht, 2000, Khan et al., 1992).
All the aforementioned studies were focused on resveratrol's chemopreventive effects. The present study attempts to understand the expression of Phase I and Phase II enzymes such as CYP1A1, CYP2E1, NAD(P)H:quinone oxidoreductase (NQO1), glutathione S-transferase (GST) and UDP-glucuronosyl transferase (UGT) as well as the expression of stress related genes upon administration of high doses of resveratrol in vivo. To study gene expression we made use of cDNA array technology, which can be a powerful and versatile method to investigate gene expression profiles.
Section snippets
Animals and treatments
CD rats (6–7 weeks old) were obtained from Charles River Laboratories. The male rats weighed approximately 175–225 g and females weighed 125–175 g at dosing initiation. The rats were housed singly in an AAALAC-accredited facility (University of Illinois at Chicago) in a temperature of 64 –79°F and humidity of 50 ± 20% with a 14 h light/10 h dark cycle. The rats were divided into four groups per sex (five rats per group). The groups were dosed with vehicle, 0.3 (dose 1), 1.0 (dose 2) or 3.0
Effect of resveratrol on liver and kidney function
After administration of three different doses of resveratrol for a period of 28 days, the rats were tested for liver and kidney function by measuring various parameters which included alanine transaminase (ALT), alkaline phosphatase (ALKP), total bilirubin (TBILI), blood urea nitrogen (BUN) and creatinine (CREAT) levels. From Table 1 it can be observed that ALT and ALKP levels were significantly elevated among male and female rats accompanied by increased total bilirubin levels among female
Discussion
This is the first study that utilizes cDNA array analysis to detect gene expression changes by in vivo administration of different doses of resveratrol to rats. A study reported an in vivo experiment, where male Sprague Dawley rats were treated with 20 mg/kg/day of trans-resveratrol for 28 days (Juan et al., 2002). In that study, it was observed that the dose of resveratrol administered was not toxic to the rats and they did not demonstrate any adverse effects on growth and development. The
Acknowledgements
This work was supported in part by NCI Contract N01-CN-95132. We thank Dr. Paul Thomas and Dr. Vladimir Mishin for technical assistance, Dr. Ron Hart and Donna Wilson for the use of real-time PCR and the members of the Kong laboratory for their critical reading of this manuscript.
References (34)
- et al.
Regioselective and stereospecific glucuronidation of trans-and cis-resveratrol in human
Arch Biochem Biophys
(2001) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
Anal Biochem
(1976)- et al.
Resveratrol is a selective human cytochrome P450 1A1 inhibitor
Biochem Biophys Res Commun
(1999) DT-Diaphorase
Methods. Enzymol.
(1967)- et al.
Inhibition of human LDL oxidation by resveratrol
Lancet
(1993) - et al.
Glutathione S-transferases. The first enzymatic step in mercapturic acid formation
J Biol Chem
(1974) - et al.
The daily oral administration of high doses of trans-resveratrol to rats for 28 days is not harmful
J Nutr
(2002) - et al.
Direct measurement of NAD(P)H:quinone reductase from cells cultured in microtiter wells: a screening assay for anticarcinogenic enzyme inducers
Anal Biochem
(1988) - et al.
Direct fluorometric methods for measuring mixed function oxidase activity
Methods Enzymol
(1978) - et al.
Anti-proliferative effect of resveratrol, a natural component of grapes and wine, on human colonic cancer cells
Cancer Lett
(2000)
Resveratrol inhibits cyclooxygenase-2 transcription and activity in phorbol ester-treated human mammary epithelial cells
J Biol Chem
Resveratrol causes WAF-1/ p21-mediated G(1)-phase arrest of cell cycle and induction of apoptosis in human epidermoid carcinoma A431 cells
Clin Cancer Res
Resveratrol causes arrest in the S-phase prior to Fas-independent apoptosis in CEM-C7H2 acute leukemia cells
Cell Death Differ
Differential inhibition and inactivation of human CYP1 enzymes by trans-resveratrol: evidence for mechanism-based inactivation of CYP1A2
J Pharmacol Exp Ther
Inhibition of aryl hydrocarbon-induced cytochrome P-450 1A1 enzyme activity and CYP1A1 expression by resveratrol
Mol Pharmacol
Transcriptional control of glutathione S-transferase gene expression by the chemoprotective agent 5-(2-pyrazinyl)-4-methyl-1,2- dithiole-3-thione (oltipraz) in rat liver
Cancer Res
Cancer chemopreventive potential of sulforamate, a novel analogue of sulforaphane that induces phase 2 drug-metabolizing enzymes
Cancer Res
Cited by (109)
Plants-based medicine implication in the evolution of chronic liver diseases
2023, Biomedicine and PharmacotherapyTargeting ERK signaling pathway by polyphenols as novel therapeutic strategy for neurodegeneration
2018, Food and Chemical ToxicologyCitation Excerpt :Moreover, acute consumption of resveratrol was found safe and well tolerated up to the concentration of 5 g/day, yet mild to moderate side effects were recorded (Patel et al., 2011). In a toxicogenomic study in rats, resveratrol did not induce satisfactory changes in the gene expression but at higher concentration, resveratrol implicated significant toxicity in the rats liver genes (Hebbar et al., 2005). Most studies stated that resveratrol does not show much adverse effects and is properly well tolerated.
Pharmacokinetics and pharmacodynamics of mycophenolic acid in Nagase analbuminemic rats: Evaluation of protein binding effects using the modeling and simulation approach
2015, Drug Metabolism and PharmacokineticsCitation Excerpt :The expression level of each mRNA was quantified by measuring the fluorescence intensity using the StepOnePlus real-time PCR system (Applied Biosystems) and was expressed as a ratio of GAPDH. The primer designs were referenced from various published articles [15–21], while the primer sequences and PCR product sizes were confirmed using primer-BLAST (http://www.ncbi.nlm.nih.gov/tools/primer-blast/). Liver microsomes from control rats and NARs were prepared according to a commonly used procedure [22].
Wine, alcohol and pills: What future for the French paradox?
2015, Life SciencesEffect of phytochemical intervention on dibenzo[a,l]pyrene-induced DNA adduct formation
2015, Mutation Research - Fundamental and Molecular Mechanisms of MutagenesisCitation Excerpt :DBP (10 μM) and NADPH (2.5 mM) were then added to the reaction mixture. The plate was read immediately at 37° C for 30 min spectrofluorometrically at an excitation of 530 nm and an emission of 580 nm for formation of resorufin [21,22]. The plate and contents were disposed as carcinogen waste.