Response of antioxidant system in rats to dietary fat and physical activity

The influence of feeding hydrogenated fat (HF) or refined peanut oil (PO) diet and regular swimming exercise on hepatic and skeletal muscle antioxidant enzymes i.e., catalase ,and glutathione peroxidase (GPX) as well as tissue lipid peroxidation was investigated in male Wistar rats. Two groups of rats were fed diet with HF or PO as the only fat source. Both the groups were further divided into 4 subgroups each according to physical activity: Two each for sedentary (HFS3, POS3) and two for swimming, HFE3 and POE3 [30 minutes a day, 6 days a week, for 3 months or HFS6, POS6, HFE6 and POE6 for 6 months. A mild increase in lipid peroxidation was observed in both liver and muscle tissues of PO-diet fed rats of E1. Swimming augmented further the lipid peroxidation in liver. GSH level was decreased in the liver of exercising rats, in contrast, it was increased in skeletal muscle by 70% in POE6 and 26% in HFE6. Compared to POS3 swimming elevated GPX activity of about 70% in liver from POE3 as well as about 60% in skeletal muscle from POE3 and POE6. The catalase activity was enhanced in muscle of HFE3 and POE3 by 250% while it remained unaltered in rats of 6 months. These data indicate an adaptive-response of antioxidant enzymes in liver and skeletal muscle to reduce oxidative stress induced by unsaturated fat (PO) and exercise.     

Antioxidant defense system in rats simultaneously intoxicated with agrochemicals

The effect of dimethoate, zineb and glyphosate administered alone or in combination on liver, kidney, brain and plasma antioxidant defense system was investigated. Lipid peroxidation, and RNS production were increased in all tissues studied, especially in those groups that received a combination of drugs. Intoxicated rats exhibited lower antioxidant ability, higher oxidized protein and glutathione levels in plasma with a decreased concentration of α-tocopherol in brain and liver, between 30% and 60% of control. Superoxide dismutase was decreased in liver and brain. Glutathione reductase was inhibited in liver while glutathione peroxidase and transferase were unaffected. Plasma lactate dehydrogenase and γ-glutamyl transpeptidase activities were both increased. The associations of drugs produce more damage than individual administration being the effects observed strongly dependent on the kind of tissue analyzed. In conclusion, the present paper evidenced both the role of the oxidative stress as a mechanism of action of some pesticides and the potential additive effects of a simultaneous exposure to more than one compound. In addition, results suggest a potential contribution of pesticide mixtures to the aetiology of some neurodegenerative diseases.     

Modulation of postprandial glycaemia and insulinaemia by dietary fat.

The present study was designed to examine the effect of corn oil (Co) on postprandial glycaemia and insulinaemia when ingested with glucose (G), casein (Cs), cellulose (Cl) and pectin (P) in various combinations. The study was conducted on six healthy male volunteers, on each of whom six meal tolerance tests were performed. The meals were isocaloric and consisted of G; G and Co; G, Co and Cs; G, Co and P; G, Co, Cs and P; and G, Co, Cs and Cl. The meals were administered after an overnight fast. In addition to a fasting blood sample, blood was collected 0.5, 1.0, 1.5 and 2.0 h after ingestion for measurement of serum glucose and insulin levels. The glycaemic response to GCo was comparable to that to G, but the insulinaemic response was significantly lower. The glycaemic response to GCoCs was significantly lower than that to G but the insulinaemic response to both was comparable. The cellulose containing meal GCoCsCl showed a further reduction in the glycaemic response but not in the insulinaemic response. The pectin containing meals GCoP and GCoCsP gave the lowest glycaemic and insulinaemic responses, the responses to the latter being lower. Corn oil by itself has only a modest effect on the postprandial metabolic response to glucose. Addition of protein and fibre, specially pectin, leads to significant attenuation of glycaemic and insulinaemic responses.     

Modulation of antioxidant defense system by the environmental fungicide carbendazim in Leydig cells of rats

Carbendazim (methyl-2-benzimidazole carbamate, MBC) a metabolite of benomyl is one of the most widespread environmental contaminant of major concern to human and animal reproductive health. The present investigation was undertaken to study the impact of carbendazim exposure on Leydig cell functions. Adult albino male rats of the Wistar strain were administered with carbendazim (25 mg/(kg (body weight)/day)) orally for 48 days. The control animals received vehicle (corn oil) alone. Another group of rats were treated with carbendazim and the same was withdrawn for a further period of 48 days. After the treatment period, rats were euthanized and blood was collected for the assay of serum hormones such as luteinizing hormone (LH), prolactin (PRL), testosterone and estradiol. Testes were immediately removed and Leydig cells were isolated in aseptic condition. Purified Leydig cells were used for quantification of steroidogenic enzymes such as 3β-hydroxysteroid dehydrogenase (3β-HSD) and 17β-hydroxysteroid dehydrogenase (17β-HSD). Leydig cellular enzymatic antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST), γ-glutamyl transpeptidase (γ-GT), glucose-6-phosphate dehydrogenase (G6PDH) and non-enzymatic antioxidants such as reduced glutathione (GSH), -tocopherol (vitamin E), ascorbic acid (vitamin C) and β-carotene (vitamin A) were assayed. Lipid peroxidation (LPO) and reactive oxygen species (ROS) were also quantified. Carbendazim exposure had no effect on body weight, serum LH and prolactin. However, testis weight, serum testosterone and estradiol were significantly decreased. In addition to this, Leydig cellular activities of steroidogenic enzymes such as 3β-HSD, 17β-HSD, antioxidant enzymes SOD, CAT, GPx, GR, GST, γ-GT, G-6-PDH and non-enzymatic antioxidants such as GSH, vitamins E, C and A were significantly diminished, whereas LPO and ROS were markedly elevated. All these above-mentioned parameters from the animals after withdrawal of MBC treatment were similar to those of the control group. Thus, the present study suggests that chronic low dose treatment of MBC is capable of inducing reproductive toxicity through increased oxidative stress, but is transient and reversible upon withdrawal of treatment.     

Effects of combined vanadate and magnesium treatment on erythrocyte antioxidant defence system in rats

The effect of vanadate and magnesium treatment on erythrocyte defence system was studied in outbred 2-month-old, albino male Wistar rats (14 rats/each group) which daily received: Group I (Control)-deionized water to drink; Group II-water solution of sodium metavanadate (NaVO(3); SMV) at a concentration of 0.125mgV/mL; Group III-water solution of magnesium sulfate (MgSO(4); MS) at a concentration of 0.06mgMg/mL, Group IV-water solution of SMV-MS at the same concentrations over a 12-week time. The fluid intake and the concentration of reduced glutathione (GSH) as well as the activity of Cu, Zn-superoxide dismutase (Cu, Zn-SOD), catalase (CAT) and glutathione reductase (GR) were significantly decreased in the rats receiving SMV alone (Group II) or in combination with MS (Group IV) compared with Groups I and III. The cellular glutathione peroxidase (cGSH-Px) activity was unchanged in all the treated groups. The activity of glutathione S-transferase (GST) fell in the animals in Group II, compared with the rats in Groups I, III and IV; whereas in the rats in Group III its activity was higher than in the control animals. These results showed that V (as SMV) consumed by the rats with drinking water at a dose of 12mgV/kg b.w./24h for 12 weeks may attenuate defence system in rats' erythrocytes (RBCs), which is probably a consequence of vanadium pro-oxidant potential. Therefore, reactive oxygen species (ROS) are suggested to be involved in the alterations in antioxidant defence system in these cells. Mg (as MS) at the dose ingested (6mgMg/kg b.w./24h) at co-exposure to SMV was not able to counteract its deleterious effect. The results also provide evidence that V-Mg interactions may be involved in the decrease of erythrocyte GR activity and Mg concentration in the plasma under concomitant treatment with both metals at the doses of 12.6mgV and 6mgMg/kg b.w./24h.     

Excessive dietary selenium decreases the vitamin A storage and the enzymatic antioxidant defence in the liver of rats

This study investigated the influence of selenium intake, over 8 weeks, on vitamin A level and on enzymatic antioxidant defence in the liver of young rats. Deficient animals were fed a well-balanced diet but without selenite addition; the Se content of this diet which originated from natural Se content of ingredients was 0.05 mg/kg. Controls were fed the same diet with 0.40 mg/kg added Se. The two other groups received high levels of Se, 2.05 or 4.05 mg/kg. Excessive Se intake decreased the concentrations of retinol and retinyl palmitate in the liver. The linear regression analysis indicated a significant (P < 0.001) dose-dependent vitamin A decline. As expected, Se deficit lowered glutathione peroxidase activity. The highest Se excess decreased the enzymatic antioxidation: Zn,Cu Superoxide dismutase, catalase, glutathione peroxidase activities. Data showed that high dietary Se can sometimes enhance carcinogenesis and our results suggest that it is best to be cautious in administrating Se to humans with the aim of preventing diseases.     

Anti-ulcerogenic effect of chitin and chitosan on mucosal antioxidant defence system in HCl-ethanol-induced ulcer in rats

The anti-ulcerogenic effect of chitin and chitosan against ulcer induced by HCl-ethanol in male Wistar rats was studied. Levels of acid output, pepsin, protein, lipid peroxides and reduced glutathione and the activity of glutathione peroxidase (GPx), glutathione-S-transferase (GST), catalase (CAT) and superoxide dismutase (SOD) were determined in the gastric mucosa of normal and experimental groups of rats. A significant increase in volume and acidity of the gastric juice was observed in the ulcer-induced group of rats. Peptic activity was significantly decreased as compared with that of normal controls. In the rats pre-treated with chitin and chitosan 2% along with feed, the volume and acid output and peptic activity of gastric mucosa were maintained at near normal levels. The level of lipid peroxidation was significantly higher in the ulcerated mucosa when compared with that of normal controls. This was paralleled by a decline in the level of reduced glutathione and in the activity of antioxidant enzymes like GPx, GST, CAT and SOD in the gastric mucosa of ulcer-induced rats. Also, the levels of mucosal proteins and glycoprotein components were significantly depleted in ulcerated mucosa. The pre-treatment with chitin and chitosan was found to exert a significant anti-ulcer effect by preventing all the HCl-ethanol-induced ulcerogenic effects in experimental rats.     

Modulation of N-nitrosomethylurea induced mammary tumorigenesis by dietary fat and voluntary exercise.

The effect of dietary fat and exercise on N-nitrosomethylurea [NMU:CAS:684-93-5]-induced mammary tumorigenesis in female F344 rats was investigated. Rats were fed the NIH-07 diet until NMU administration on day 50 of age, when they were transferred to four treatment groups. Three sedentary groups were fed either high-fat (20% wt/wt), medium fat (10%) or low fat (5%) diets (HF, MF, LF, respectively), and a fourth group was fed a HF diet but allowed free access to an activity wheel (HFEX). Tumor yields among the three sedentary groups were significantly greater in the HF and MF groups when compared to the LF group. Voluntary exercise reduced tumor yields and delayed time of tumor appearance in HFEX animals to levels similar to those found in LF sedentary animals. Animals with voluntary access to exercise wheels averaged between 1.03 and 2.85 miles/day, consumed more food (+ 18%) and exhibited greater weight gain (+ 13%) than their sedentary counterparts. No differences in weight gains were detected among the HF, MF, and LF groups, despite widely varying amounts of fat intake. Body composition studies indicated that body fat content was not influenced by the quantity of fat consumed in the diet, but was significantly reduced by voluntary exercise (-20%). Since exercise and fat intake have been associated with alterations in endocrine status, circulating bioactive and immunoactive prolactin were assessed at termination. No significant changes were found in either form of prolactin among the four experimental groups, casting doubt on mediation by this pituitary hormone.     

Modulation of melanogenesis and antioxidant defense system in melanocytes by amikacin

Amikacin is principally used to treat infections caused by microorganisms resistant to other aminoglycosides. Ototoxicity is one of the side effects of amikacin, but the causative mechanism of damage to the ear has not been fully established. Thus, the aim of this work was to examine the impact of amikacin on the melanogenesis and antioxidant defense system in cultured human normal melanocytes (HEMa-LP). Amikacin induced the concentration – dependent loss in melanocytes viability. The value of EC₅₀ was determined to be ～7.5 mM. The analyzed antibiotic inhibited melanin biosynthesis in concentration-dependent manner. Increasing the amikacin concentration also resulted in a decrease in cellular tyrosinase activity. To study the antioxidant defense system in melanocytes, the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in cells exposed to amikacin were determined. Significant changes in cellular antioxidant enzymes activities were observed. Modulation of melanogenesis and the antioxidant status of melanocytes resulting from the use of amikacin in vitro may explain a potential role of melanin and melanocytes in the mechanisms of aminoglycosides ototoxic effects in vivo.     

Failure of recovery from lead induced hepatoxicity and disruption of erythrocyte antioxidant defence system in Wistar rats

Lead acetate (PbA) is one of the major environmental contaminants with grave toxicological consequences both in the developing and developed countries. The liver and erythrocyte antioxidant status and markers of oxidative were assessed. Exposure of rats to PbA led to significant decline (p < 0.05) in hepatic and erythrocyte glutathione peroxidase (GPx), glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), and reduced glutathione (GSH) content. Similarly, malondialdehyde (MDA) and H₂O₂ concentrations were significantly (p < 0.05) elevated. Histopathology and immunohistology of liver of rats exposed to PbA showed focal areas of necrosis and COX-2 expression after 6 weeks of PbA withdrawal. Taken together, hepatic and erythrocytes antioxidant defence system failed to recover after withdrawal of the exposed PbA for the period of the study. In conclusion, experimental animals exposed to PbA did not recover from hepatotoxicity and disruption of erythrocyte antioxidant defence system via free radical generation and oxidative stress.     

Brain regional responses in antioxidant system to alpha-lipoic acid in arsenic intoxicated rat

Impaired antioxidant defense mechanisms and oxidative stress are implicated in the pathogenesis of arsenic toxicity. Our study was designed to determine whether alpha-lipoic acid, which has been shown to have substantial antioxidant properties, when administered (70 mg/kg body weight) once daily for 60 days along with arsenic (100 ppm sodium arsenite mixed in drinking water) would prevent arsenic-induced changes in antioxidant defense system, superoxide dismutase (SOD-total SOD, Mn SOD, Cu/Zn SOD), catalase (CAT) and glutathione peroxidase (GSH-PX) in rat brain regions such as cortex, hypothalamus, striatum, cerebellum and hippocampus. The present study also examined the effect of alpha-lipoic acid over arsenic-induced oxidant production and lipid peroxidation level (LPO) in discrete brain regions of rats. The cortex, striatum and hippocampus showed greater decreases in GSH-Px enzyme activity than cerebellum and hypothalamus with arsenic exposure. Striatum had the greatest percentage of decreased activities of total SOD and Mn SOD, whereas cortex had the greatest percentage decrease in the activity of Cu/Zn SOD in arsenic-alone treated rats. Hypothalamus and cerebellum exhibited the lowest catalase activity among all tested regions in arsenic-only treated rats. Rate of dichlorofluorescin oxidation, an indication of reactive oxygen species and other intracellular oxidants production was increased with arsenic exposure in all brain regions studied. Cortex, hippocampus and striatum exhibited greater increase of LPO levels than cerebellum and hypothalamus. SOD, CAT, GSH-Px activities were upregulated in arsenic plus lipoic acid treated versus arsenic-only treated rats. Also, simultaneous lipoic acid treatment along with arsenic proved to be sufficient in reducing oxidant production and LPO level in all rat brain regions. Our results demonstrate that arsenic-induced deficits in antioxidant enzyme activities and increase in oxidant production and lipid peroxidation level in brain regions can be overcome through simultaneous treatment with lipoic acid.     

Effect of dietary fat upon ethanol metabolism in rats

The effect of dietary fat upon ethanol metabolism was studied in rats. Wistar strain male rats were divided into four groups according to diet, namely alcohol-high fat, alcohol-low fat, control-high fat, and control-low fat. After 4 weeks of feeding, blood ethanol levels following an intraperitoneal injection of 0.2 g ethanol/100 g of body weight were measured. The disappearance rate of blood ethanol was faster and the metabolic rate of ethanol was significantly greater in the alcohol-high fat group compared to the alcohol-low fat or non-alcoholic groups. Microsomal enzymes, such as the microsomal ethanol-oxidizing system, aniline hydroxylase. and glucose-6-phosphatase, were significantly higher in the alcohol-high fat group than in the alcohol-low fat or non-alcoholic groups. The ethanol uptake rate of the isolated perfused liver was increased significantly in the alcoholic groups. In the alcoholic rats, the high fat group showed significantly higher uptake than the low fat group. Although the ethanol uptake rate after 4-methylpyrazole treatment was not significantly different among the four groups, its fraction of the total ethanol uptake was increased significantly in the alcohol-high fat group. These results suggest that high fat diets accelerate ethanol metabolism through the microsomal ethanoloxidizing system.     

3－硝基丙酸染毒大鼠的神经病理变化

用２０，４０和８０ｍｇ／ｋｇ三种剂量的３－硝基丙酸灌胃染毒２４只大鼠，观察中毒后神经病理改变。结果表明：３－硝基丙酸的主要损伤部位是尾－壳核。电镜检查可见神经元胞浆疏松，，核染色质积聚，最后胞浆和核均发生固缩。髓鞘变薄或断裂，轴索膜与髓鞘间水肿。星形胶质细胞水肿，血管周围有肿胀的突起，此与缺血性脑病理改变无明显差别。     

Alpha-tocopherol and alpha-lipoic acid enhance the erythrocyte antioxidant defence in cyclosporine A-treated rats

The aim of this study was to determine the effects of dietary antioxidant supplementation with alpha-tocopherol and alpha-lipoic acid on cyclosporine A (cyclosporine)-induced alterations to erythrocyte and plasma redox balance. Rats were randomly assigned to either control, antioxidant (alpha-tocopherol 1000 IU/kg diet and alpha-lipoic acid 1.6 g/kg diet), cyclosporine (25 mg/kg/day), or cyclosporine + antioxidant treatments. Cyclosporine was administered for 7 days after an 8 week feeding period. Plasma was analysed for alpha-tocopherol, total antioxidant capacity, malondialdehyde, and creatinine. Erythrocytes were analysed for glutathione, methaemoglobin, superoxide dismutase, catalase, glutathione peroxidase, glucose-6-phosphate dehydrogenase, alpha-tocopherol and malondialdehye. Cyclosporine administration caused a significant decrease in superoxide dismutase activity (P<0.05 control versus cyclosporine) and this was improved by antioxidant supplementation (P<0.05 cyclosporine versus cyclosporine + antioxidant; P<0.05 control versus cyclosporine + antioxidant). Animals receiving cyclosporine and antioxidants showed significantly increased (P<0.05) catalase activity compared to both groups not receiving cyclosporine. Cyclosporine administration induced significant increases in plasma malondialdehyde and creatinine concentration (P<0.05 control versus cyclosporine). Antioxidant supplementation prevented the cyclosporine induced increase in plasma creatinine (P<0.05 cyclosporine versus cyclosporine + antioxidant; P>0.05 control versus cyclosporine + antioxidant), however, supplementation did not alter the cyclosporine induced increase in plasma malondialdehyde concentration (P>0.05 cyclosporine versus cyclosporine + antioxidant). Antioxidant supplementation resulted in significant increases (P<0.05) in plasma and erythrocyte alpha-tocopherol in both of the supplemented groups compared to non-supplemented groups. In conclusion, dietary supplementation with alpha-tocopherol and alpha-lipoic acid enhanced the erythrocyte antioxidant defence and reduced nephrotoxicity in cyclosporine treated animals.     

Effect of Silybum marianum oil and legalon on lipid peroxidation and liver antioxidant systems in rats intoxicated with carbon tetrachloride

An oil obtained from the seeds of Saint-Mary thistle (Silybum marianum) and the drug legalon (silybinin) prepared from this plant produce an antioxidant effect on liver tissues of rats poisoned with carbon tetrachloride. Legalon (25 mg/kg) and the oil (2000 mg) reduced the level of lipid peroxidation, increased the catalase activity, but did not reduce the concentration of selenium in liver (which decreased as a result of CCl4 intoxication). Legalon significantly increased the activity of superoxide dismutase in liver tissues, while the Saint-Mary thistle oil did not produce such effect.     

Aspirin disposition in rats acutely intoxicated with CCl4

The profile of urinary salicylate metabolites was determined after an oral administration of acetylsalicylic acid (ASA) to: 1, control rats; 2, rats treated with CCl4 and 3, rats intoxicated with CCl4 and also pretreated with colchicine for 7 days. The following enzymatic activities were determined: liver and plasma ASA-esterase, liver UDP-glucuronyltransferase and liver aniline hydroxylase. The time course of plasma concentration of salicylates in similar groups were followed after the intraperitoneal administration of acetylsalicylic acid (ASA), salicylic acid (SA) or gentisic acid (GA). The animals acutely intoxicated with CCl4 showed a reduction in urinary excretion of glucuronates and an increased urinary excretion of gentisic and salicylic acids. The activities of plasma and liver ASA-esterases were significantly increased in CCl4-treated rats while the aniline hydroxylase was reduced and the UDP-glucuronyltransferase remained unchanged. The plasma half lives of salicylates were reduced in CCl4-treated rats regardless of the administered parent compound. Colchicine pre-treatment completely prevented the alterations produced by acute intoxication with CCl4. The heterogeneity of liver metabolic dysfunctions present in acute liver damage was evidenced. It is emphasized that the pharmacokinetic alterations produced by acute liver injury can be the result of complex factors that may involve changes in circulation, hepatic binding protein and other routes of elimination.     

Preference for drinking water containing dimercaptosuccinic acid by rats intoxicated with methylmercury

Rats made anorexic by methylmercury treatment preferred aqueous, 2,3-dimercaptosuccinic acid (2.5 mg DMSA/ml) to tap water. Food and fluid consumption or the clearance of mercury increased approximately to the same extent when both aqueous DMSA and tap water or only aqueous DMSA was available. Methylmercury-intoxicated rats did not show the same preference for -cysteine and untreated rats had no preference for aqueous DMSA.     

Neurofilament degradation in the nervous system of rats intoxicated with acrylamide,related compounds or 2,5-hexanedione

Degradation of neurofilament (NF) proteins by Ca²⁺-activated neutral protease (CANP) was studied in the nervous system of rats treated with neurotoxic or non-neurotoxic compounds. In the tibial nerve, the degradation of NF 68K was depressed by five neurotoxic compounds: acrylamide, N-hydroxymetylacrylamide, N-isopropylacrylamide, methacrylamide and 2,5-hexanedione. A non-neurotoxic compound, diacetone acrylamide, did not show any effect on the degradation. An immunoblot analysis confirmed the reduction in the degradation and revealed a difference in the degradation pattern between the control and acrylamide-treated rats. In the spinal cord, the degradation of the three subunits of NF was depressed in animals treated with acrylamide. Although the exact mechanism of the reduction in the degradation of NF is not yet known, the present results suggest that an inhibitory effect on CANP activity might be relevant to the mechanism of neurotoxic action of acrylamide derivatives.Abbreviations CANP calcium-activated neutral protease - CBB Coomassie Brilliant Blue R-250 - NF neurofilament - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis     

Biliary secretory function in rats chronically intoxicated with aluminum.

The effects of a chronic aluminum (Al) exposure on biliary secretory function, with special emphasis on hepatic handling of non-bile salt organic anions, was investigated. Male Wistar rats received, intraperitoneally, either 27 mg/kg body weight of Al, as Al hydroxide [Al (+) rats], or the vehicle saline [Al (-) rats] three times a week for 3 months. Serum and hepatic Al levels were increased by the treatment (approximately 9- and 4-fold, respectively). This was associated with enhanced malondialdehyde formation (+110%) and a reduction in GSH content (-17%) and in the activity of the antioxidant enzymes catalase (-84%) and GSH peroxidase (-46%). Bile flow (-23%) and the biliary output of bile salts (-39%), cholesterol (-43%), and proteins (-38%) also decreased. Compartmental analysis of the plasma decay of the model organic anion bromosulphophthalein revealed that sinusoidal uptake and canalicular excretion of the dye were significantly decreased in Al (+) rats (-53 and -43%, respectively). Expression of multidrug resistance-associated protein 2 (Mrp2), the main, multispecific transporter involved in the canalicular excretion of organic anions, was also decreased (-40%), which was associated with a significant decrease in the cumulative biliary excretion of the Mrp2 substrate, dinitrophenyl-S-glutathione (-50%). These results show that chronic Al exposure leads to oxidative stress, cholestasis, and impairment of the hepatic handling of organic anions by decreasing both sinusoidal uptake and canalicular excretion. The alteration of the latter process seems to be causally related to impairment of Mrp2 expression. We have addressed some possible mechanisms involved in these deleterious effects.