[771] INDUCTION LIVER DISEASE IN COMBINED ADMINISTRATION ACETAMINOPHEN AND ETHANOL

OK. ALCOHOLIC LIVER DISEASE, NAFLD AND DRUC-INDUCED LIVER DISEASE were to define the relationship between ferritin and iron stores in patients with NAFLD, the effect of iron depletion on insulin resistance, and whether basal ferritin levels influence treatment outcome. Methods: Subjects were included if ferritin and/or ALT were persistently elevated after 4 months of standard therapy. Sixty-four phlebotomized subjects were matched 1:l for age, sex, ferritin, obesity, and ALT levels with patients who underwent lifestyle modifications only. Insulin resistance was evaluated by insulin levels, determined by RIA, and the HOMA-R index, at baseline and after 8 months. Results: Baseline ferritin levels were associated with body iron stores (p=0.006). Iron depletion resulted in a significantly larger decrease in insulin resistance (p = 0.0002 for insulin, p = 0.0007 for HOMA-R) compared with nutritional counseling alone, independently of changes in BMI, baseline HOMA-R, and the presence of the metabolic syndrome. Iron depletion was more effective in reducing HOMA-R in patients in the top two tertiles offerritin concentrations (p 0.05 vs. controls), but not in the first tertile, and in carriers of mutations in the HFE gene of hereditary hemochromatosis vs. non-carriers (p < 0.05). Conclusions: Given that phlebotomy reduces insulin resistance, which correlates with tissue damage, future studies should evaluate the effect of iron depletion on liver histology and cardiovascular endpoints. I

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(p i 0.05), and with its inhibitor, the transcription factor FoxO 1 (p=. I , ns). At the same time, Akt2 relative activity and lipogenic factors were higher in NASH vs. controls (p 0.001). The TRB3 84Arg allele, determining increased activity, was associated with lower steatosis percentage ( I 7*20 vs. 36&22; p=.01) and Fox01 expression (p=.01), and with increased insulin sensitivity (p = .05) in patients with NAFLD.

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Conclusions: Decreased TRB3 expression is involved in lipogenesis and insulin resistance in NAFLD, and it could be related to overexpression and increased activity of FoxO I .

17711 ADMINISTRATION INDUCTION LIVER DISEASE IN COMBINED ACETAMINOPHEN AND ETHANOL A.K. Voronina, A.M. Shayakhmetova, VN. Kovalenko. Department qf General Toxicology, Institute of Pharmacology and Toxicology, Acadenzy of Medical Sciencex of Ukraine, Kviu, Ubaine E-mail: [email protected]

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Effect of phlebotomy on insulin resistance (as detected by the HOMA-IR index) according to ferritin tertiles. White bars: phlebotomized subjects; grey bars: controls.

17701 ROLE OF TRB3 IN THE REGULATION OF HEPATIC LIPOGENESIS IN NONALCOHOLIC FATTY LIVER DISEASE L. Valenti, R. Rametta, P. Dongiovanni, A.L. Fracanzani, S. Fargion. Department of Internal Medicine, Uniuersita ' di Milano, O~yedale Policlinico IRCCS, Milano, Italy E-mail: 1uca.valentil~junimi.it Background and Aims: The Drosophila Tribbles homolog TRB3 is a psuedokinase which inhibits lipogenesis by inactivating the kinase Akt, which mediates the insulin drive on lipid metabolism, and by promoting ubiquitination and degradation of acetyl-coenzyme A carboxylase, rate limiting enzyme in lipid synthesis. Aim was to evaluate TRB3 expression in patients with NAFLD. Methods: Liver biopsies of 20 patients with NAFLD (10 NASH according do Kleiner), and 15 without metabolic disease were considered; gene expression measured by qRT-PCR and Western blotting; the TRB3 Glu84Arg polymorphism evaluated by restriction analysis. Results were compared by ANOVA and post-hoc analysis, and correlated by linear regression. Results: Patients with NASH had higher steatosis percentage, the triglycerides/HDL ratio (TgVHDL), and Homa insulin resistance index (Homa0.05). TRB3 expression was lower IR) compared to other groups (p i in NASH compared to simple steatosis and controls ( p = .05), and was negatively correlated with steatosis percentage, TgllHDL, and HOMA-IR

Background and Aims: It is known that P-4502E1 metabolizes acetaminophen, and that it can be induced after by inducers such as ethanol. The purpose of this study was to clarity the effects of ethanol ingestion and chronic ethanol consumption on the hepatotoxicity of acetaminophen. Methods: Effect of combined administration of both acetaminophen and ethanol on aniline-N-hydroxylase activity and malonodialdehyde (MDA) formation, contents of glutathione and protein SH-groups, liver glutathione reductase and catalase activity, alanine aminotransferase (ALT) activity was studied. After blood collection for serum enzymes, the liver tissue was collected, washed with cold saline, blotted dry and was homogenized. Preparation hepatic microsomes: the livers were removed, quickly cooled on ice and homogenized. Results: Preliminary studies in mice support the concept that acetaminophen hepatotoxicity may be enhanced by ethanol ingestion. A significant reduction in the LD50 was seen in the alcohol-treated mice. In treated ethanol rats (3.2mglkg b. w.), the activities of aniline-Nhydroxylase (a marker of cytochrome P-450 2E1 activity) in liver microsomes significant increased after acetaminophen (dose 400 mgikg bw) administration compared with control animals. Among these glutathione and protein SH-groups contents as well as liver glutathione reductase and catalase activity were decreased. The rate of induced MDA formation, hydroperoxides and super oxide anion contents were increased in comparison to the control. ALT activity elevated in blood serum of 1.7 fold the upper limit of normal. After chronic ethanol consumption, anilineN-hydroxylase, glutathione reductase, ALT activities and MDA formation was increased significantly, whereas content of glutathione were decreased compared with control. After parallel administration of acetaminophen (dose 500 mg/kg) the activities of aniline-N-hydroxylase followed up 65% and ALT activities increased of 5.6 fold. In acetaminophen-treated animals, ethanol-fed rats showed a significant decreased hepatic glutathione levels. Induction of CYP2EI by chronic alcohol ingestion markedly increases the susceptibility to the toxicity of acetaminophen. Conclusions: Our results suggest that acetaminophen and alcohol intensify the induction of CYP2EI in a synergistic manner may help to understand and to prevent an additional risk factor for developing acetaminophen hepatotoxicity. ~

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