Oxidative stress mediates drug-induced hepatotoxicity in rats: a possible role of DNA fragmentation
Introduction
Hepatic DNA fragmentation has been reported in many hepatotoxicity models following treatments with drugs such as acetaminophen (Ray et al., 1990), dimethylnitrosamine (Ray et al., 1992), carbon tetrachloride (Shi et al., 1998) and diquat (Gupta et al., 2000).
DDC is one of the dithiocarbamate compounds that are commonly used in industry, agriculture, and medicine (Cheng and Trombetta, 2004). In addition, DDC is considered as a radio protective agent (Fuentes et al., 1998) as well as immunotherapy for breast cancer (Dufour et al., 1993). DDC is also receiving more attention as a useful medication in treating acquired immunodeficiency syndrome (AIDS) due to its immune modulating action and antiviral activity against AIDS virus (Reisinger et al., 1990). Recent studies show that thiocarbamates-induced apoptosis is mediated via activation of caspase-3, decrease mitochondrial membrane potential, increase free radical production, and deplete GSH (Chen et al., 2001). Due to its ability to form a lipophilic complex, DDC can easily cross the cell membrane dramatically increasing the intracellular copper (Delmaestro and Trombetta, 1995), thereby disrupting intracellular copper homeostasis. This increase could cause oxidative stress by hydroxyl free radical production and could also interfere with copper-dependent enzyme reactions, such as Cu, Zn superoxide dismutase and many other enzymes (Simonian et al., 1992). Both hepatotoxicity and oxidative stress have been reported in rats following a single dose treatment with DDC (Ishiyama et al., 1990). In vitro, DDC-induced DNA fragmentation has also been documented (Burkitt et al., 1998).
Diclofenac (DIC) (Voltaren) is a non-steroidal anti-inflammatory agent, which has antipyretic, analgesic and anti-inflammatory effects (Aydin et al., 2003). The toxic and apoptotic effects of DIC might be involved in drug-induced renal toxicity and hepatotoxicity both in humans (Ouellette et al., 1991, Hackstein et al., 1998) and experimental animals (Aydin et al., 2003). These characteristics of DIC may also apply to a subset of anticancer agents. DIC has also been shown to induce DNA fragmentation in kidney of mice (Hickey et al., 2001) and in human and rat hepatocytes (Gomez-Lechon et al., 2003).
Ketoconazol (KET) is a new antifungal drug related to imidazol. KET is characterized by its wide spectrum of activity and oral availability which dramatically broaden its clinical applications (Ma et al., 2003). KET-induced hepatotoxicity has been documented in human (Chien et al., 2003, Kim et al., 2003), rats (Rodriguez and Buckholz, 2003) and rabbits (Ma et al., 2003).
The main objective of this report is to highlight the biochemical alterations, namely increased lipid peroxidation, decreased GSH contents and SOD activity, increased DNA fragmentation and increased intracellular calcium content, as a common feature of the DDC-, DIC- and KET-induced hepatotoxicity.
Section snippets
Chemicals
DIC was obtained from Novartis Pharma AG, Basel, Switzerland. DDC, and KET were obtained from Sigma. Reagent kits for ALT and AST enzymes were purchased from BioMerieux (Marcy–I’Etoile, France). Thiobarbituric acid, reduced glutathione, 5,5-dithiobis(2-nitrobenzoic acid) (DTNB), Folin's reagent, epinephrine, SOD enzyme, H2O2 and bovine albumin were obtained from Sigma Chemical Co. (St. Louis, MO). All other chemicals were obtained from common commercial suppliers.
Animals
Male Wistar rats (150–200 g)
Effects of DDC, DIC and KET intoxication on liver functions
After treatment with DDC, levels of serum ALT and AST has been significantly increased compared to the control by about 117 and 105%, respectively (Fig. 1). Liver injury manifested by elevation of serum ALT (283% of control) and AST (266% of control) activities was observed 24 h after DIC treatment (Fig. 1). Rats treated with acute dose of KET showed a significant increase in ALT (480% of control) and AST (236% of control) activities.
Effect of DDC, DIC and KET on oxidative stress
The effects of hepatotoxic agents on oxidative stress were
Discussion
In the present work, the severe hepatic damage induced with DDC, DIC or KET was evidenced by the elevation in liver enzymes (serum ALT and AST) and confirmed by histological alterations in the liver. Similar results have shown an increase of serum aminotransferase activities associated with damage of hepatic tissue following the treatment with DDC (Ishiyama et al., 1990), DIC (Aydin et al., 2003) or KET (Rodriguez and Buckholz, 2003) in albino rats. Many studies have attempted to clarify the
Acknowledgments
The authors are grateful to Ms. Aysha Alkaabi and Ms Dina Amer (Biology Department, UAE University) for their valuable assistance throughout this study. We are also thankful to Mr. Sayel Daoud (Twam hospital, UAE) for his kind help with histological preparations. Authors are indebted to Robert Monk, MSc (University of Pennsylvania, USA) for proofreading the manuscript.
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