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84 Prevalence of and risk factors for non-alcoholic fatty liver (NAFL) in the general population of Northern Italy
General Session 3: Molecular Hepatitis Conclusion: CRP is a highly specific marker of alcoholic hepatitis and allows to avoid liver biopsy in some alcoholics with severe alcoholic hepatitis before corticotherapy.
82 ACETYL SALICYLIC ACID AND FASTING CAUSE REYE-LIKE SYNDROME IN MICE HETEROZYGOUS FOR A MITOCHONDRIAL TRIFUNCTIONAL PROTEIN DEFECT
Z. Yang 1 , M. Cline 2 , J.A. Ibdah 3 . 1 Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, China; 2 Division of Gastroenterology, Wake Forest University School of Medicine, Winston-Salem, USA Background: The pathogenesis underlying Reye’s syndrome and its association with salicylates use remains unknown. Mitochondrial trifunctional protein (TFP) is a hetero-octamer of 4 a and 4 b subunits that catalyze the final 3 steps of long chain fatty acid oxidation. Most children with TFP deficiency present at few months of age with predominantly Reye-like manifestations and micro-vesicular hepatic steatosis, the histological hallmark of Reye’s syndrome. Heterozygotes carrying mutations in TFP are phenotypically normal and constitute @1% of the general population. Hypothesis: Heterozygosity for TFP mutations predisposes to development of Reye’s syndrome. Methods: We used gene targeting replacement strategy to generate TFP a-subunit null allele. Acetyl salicylic acid (ASA) was administered via intraperitoneal injections at doses 400 mg/kg/day, for 2 days, to one-month old +/- or +/+ mice, with and without 24-hour fasting prior to sacrifice. Routine histology and Oil O Red stain were used to assess hepatic changes. Serum transaminases were measured and correlated to the histological changes. Data were analyzed using 6 mice of each genotype. Results: Combination of ASA and fasting caused significant microvesicular hepatic steatosis without necrosis and higher serum transaminases levels in the +/- mice compared to the +/+ mice (P<0.001). ASA without fasting, and fasting alone did not cause significant changes. Our molecular and enzymatic analyses suggest that ASA directly reduces the activities of TFP enzymes without altering gene expression or protein synthesis. Conclusions: Heterozygosity for TFP mutations predisposes to development of Reye’s syndrome under conditions of fasting and salicylates use.
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liver. IkB degradation was also markedly increased in NASH and ASH (73 and 83%, respectively; p<0.01), possibly reflecting NF-kB activation. In fact, patients with steatohepatitis showed increased portal and lobular inflammation and fibrosis, compared with controls (p<0.001). Conclusions: Liver injury in NASH and ASH is associated with increased death receptor-mediated apoptosis, inflammation, and fibrosis. Further, anti-apoptotic Bcl-2 protein is strongly upregulated, probably reflecting apoptotic stress. These results might provide additional targets for therapeutic intervention. (Funded by the Portuguese Soc. of Gastroenterology)
84 PREVALENCE OF AND RISK FACTORS FOR NON-ALCOHOLIC FATTY LIVER (NAFL) IN THE GENERAL POPULATION OF NORTHERN ITALY
G. Bedogni 1 , L. Miglioli 1 , M. Passalacqua 1 , F. Cortesi 1 , N. Batrtistini 2 , C. Tiribelli 1 , S. Bellentani 1 . 1 Liver and Nutrition Center, Fondo Studi Fegato, Modena, Italy; 2 Human Nutrition Chair, University of Modena and Reggio Emilia, Modena, Italy Aims: A strong association between overweight and fatty liver (FL) is well known. In order to better define the association between nutrition and NAFL, an intra-cohort case-control study was designed, starting from the population of the still ongoing Dionysos study. Methods: The study was performed between Jan 2002 and Aug 2003 on all the adults living in the city of Campogalliano (Modena, Italy). Every subject (“case”) fulfilling the Dionysos criteria for the presence of liver disease (n = 497) was matched with a randomly chosen subject of the same age and sex but without liver disease (“control”). Besides a clinical and laboratory evaluation, the subjects underwent a liver ultrasonography, a detailed anthropometrical evaluation and a 7-day diary of food habits. Results: Compliance was 66%. Prevalence of FL was 48% in cases and 38% in controls (p = 0.726). NAFL was responsible for 45% and 56% cases of FL in cases and controls, respectively (p = 0.012). After correction for age and sex, the presence of NAFL in the pooled sample was independently associated with a body mass index ≥ 30 kg/m2 (OR = 3.1; 95%CI 1.7 to 5.5, p = 0.0002) and a with a waist circumference ≥ 88 cm in males and ≥ 102 cm in females (OR = 2.1; 95%CI 1.1 to 3.9, p = 0.017). Conclusions: Waist circumference is a risk factor for NAFL independent from BMI; many cases of NAFL will be missed if common screening criteria are used.
83 ENHANCED APOPTOSIS AND BCL-2 PROTEIN PRODUCTION IN THE LIVER OF PATIENTS WITH STEATOHEPATITIS
R.M. Ramalho 1 , H. Cortez-Pinto 2 , S. Solá 1 , R.E. Castro 1 , M.E. Camilo 2 , M.C. Moura 3 , C.M.P. Rodrigues 1 . 1 Centro De Patogénese Molecular, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal; 2 Centro De Nutrição E Metabolismo, IMM, Faculty of Medicine, University of Lisbon, Lisbon, Portugal; 3 Department of Gastroenterology, IMM, Faculty of Medicine, University of Lisbon, Lisbon, Portugal Background/Aims: Apoptosis appears to play a role in the pathogenesis of non-alcoholic (NASH) and alcoholic steatohepatitis (ASH), perhaps contributing to tissue inflammation and fibrogenesis. In this study, we investigated the modulation of apoptosis-related proteins in the liver of patients with steatohepatitis. Methods: Caspase-3 processing was evaluated by immunoblot analysis of liver tissue from 12 patients with NASH, 12 with ASH and 3 histologically normal controls. In addition, expression of death receptors, Bcl-2 family members, and NF-kB inhibitor (IkB) were determined by Western blot. Liver biopsies were also graded for inflammation and fibrosis. Results: Caspase-3 processing was >25% increased in steatohepatitis (p<0.05). Caspase activation coincided with upregulation of proapoptotic Bax (up to 3-fold, p<0.05), confirming the occurrence of apoptosis. Further, production of type I tumor necrosis factor-a receptor was increased 2and 3-fold in NASH and ASH, respectively (p<0.05); the Fas receptor was only slightly elevated. Notably, Bcl-2 expression was strongly enhanced in steatohepatitis (up to 150-fold, p<0.01), but almost absent in control
General Session 3: Molecular Hepatitis
85 ENDOGENOUS OPIOIDS INHIBIT CHOLANGIOCYTE PROLIFERATION IN THE BDL BUT NOT IN THE NORMAL RAT
M. Marzioni 1 , G. Alpini 2 , S. Saccomanno 1 , L. Trozzi 1 , S. Glaser 2 , G. Macarri 1 , A. Benedetti 1 . 1 Dept. of Gastroenterology, Università Delle Marche School of Medicine, Ancona, Italy; 2 The Texas A&M University and Scott&White Hospital HSC, Temple TX, USA In the course of cholestasis the opioid neuromodulation, opioid peptide plasma levels and activity are significantly increased. Proliferating bile ducts express the endogenous opioid precursor Met-enkephalin. Hormoneinduced changes in the Ca2+ signalling reduce cholangiocyte proliferation, interfering with MAP-kinase signalling. Aim: Are endogenous opioids able to regulate cholangiocyte proliferation? Methods: Expression of delta opioid receptor (DOR) in cholangiocytes was studied by immunohistochemistry and immunoblots in pure cholangiocytes from normal and 1 week BDL rats. Pure cholangiocytes from normal and 1 week BDL rats were incubated in vitro with the selective
82 ACETYL SALICYLIC ACID AND FASTING CAUSE REYE-LIKE SYNDROME IN MICE HETEROZYGOUS FOR A MITOCHONDRIAL TRIFUNCTIONAL PROTEIN DEFECT
Z. Yang 1 , M. Cline 2 , J.A. Ibdah 3 . 1 Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, China; 2 Division of Gastroenterology, Wake Forest University School of Medicine, Winston-Salem, USA Background: The pathogenesis underlying Reye’s syndrome and its association with salicylates use remains unknown. Mitochondrial trifunctional protein (TFP) is a hetero-octamer of 4 a and 4 b subunits that catalyze the final 3 steps of long chain fatty acid oxidation. Most children with TFP deficiency present at few months of age with predominantly Reye-like manifestations and micro-vesicular hepatic steatosis, the histological hallmark of Reye’s syndrome. Heterozygotes carrying mutations in TFP are phenotypically normal and constitute @1% of the general population. Hypothesis: Heterozygosity for TFP mutations predisposes to development of Reye’s syndrome. Methods: We used gene targeting replacement strategy to generate TFP a-subunit null allele. Acetyl salicylic acid (ASA) was administered via intraperitoneal injections at doses 400 mg/kg/day, for 2 days, to one-month old +/- or +/+ mice, with and without 24-hour fasting prior to sacrifice. Routine histology and Oil O Red stain were used to assess hepatic changes. Serum transaminases were measured and correlated to the histological changes. Data were analyzed using 6 mice of each genotype. Results: Combination of ASA and fasting caused significant microvesicular hepatic steatosis without necrosis and higher serum transaminases levels in the +/- mice compared to the +/+ mice (P<0.001). ASA without fasting, and fasting alone did not cause significant changes. Our molecular and enzymatic analyses suggest that ASA directly reduces the activities of TFP enzymes without altering gene expression or protein synthesis. Conclusions: Heterozygosity for TFP mutations predisposes to development of Reye’s syndrome under conditions of fasting and salicylates use.
29
liver. IkB degradation was also markedly increased in NASH and ASH (73 and 83%, respectively; p<0.01), possibly reflecting NF-kB activation. In fact, patients with steatohepatitis showed increased portal and lobular inflammation and fibrosis, compared with controls (p<0.001). Conclusions: Liver injury in NASH and ASH is associated with increased death receptor-mediated apoptosis, inflammation, and fibrosis. Further, anti-apoptotic Bcl-2 protein is strongly upregulated, probably reflecting apoptotic stress. These results might provide additional targets for therapeutic intervention. (Funded by the Portuguese Soc. of Gastroenterology)
84 PREVALENCE OF AND RISK FACTORS FOR NON-ALCOHOLIC FATTY LIVER (NAFL) IN THE GENERAL POPULATION OF NORTHERN ITALY
G. Bedogni 1 , L. Miglioli 1 , M. Passalacqua 1 , F. Cortesi 1 , N. Batrtistini 2 , C. Tiribelli 1 , S. Bellentani 1 . 1 Liver and Nutrition Center, Fondo Studi Fegato, Modena, Italy; 2 Human Nutrition Chair, University of Modena and Reggio Emilia, Modena, Italy Aims: A strong association between overweight and fatty liver (FL) is well known. In order to better define the association between nutrition and NAFL, an intra-cohort case-control study was designed, starting from the population of the still ongoing Dionysos study. Methods: The study was performed between Jan 2002 and Aug 2003 on all the adults living in the city of Campogalliano (Modena, Italy). Every subject (“case”) fulfilling the Dionysos criteria for the presence of liver disease (n = 497) was matched with a randomly chosen subject of the same age and sex but without liver disease (“control”). Besides a clinical and laboratory evaluation, the subjects underwent a liver ultrasonography, a detailed anthropometrical evaluation and a 7-day diary of food habits. Results: Compliance was 66%. Prevalence of FL was 48% in cases and 38% in controls (p = 0.726). NAFL was responsible for 45% and 56% cases of FL in cases and controls, respectively (p = 0.012). After correction for age and sex, the presence of NAFL in the pooled sample was independently associated with a body mass index ≥ 30 kg/m2 (OR = 3.1; 95%CI 1.7 to 5.5, p = 0.0002) and a with a waist circumference ≥ 88 cm in males and ≥ 102 cm in females (OR = 2.1; 95%CI 1.1 to 3.9, p = 0.017). Conclusions: Waist circumference is a risk factor for NAFL independent from BMI; many cases of NAFL will be missed if common screening criteria are used.
83 ENHANCED APOPTOSIS AND BCL-2 PROTEIN PRODUCTION IN THE LIVER OF PATIENTS WITH STEATOHEPATITIS
R.M. Ramalho 1 , H. Cortez-Pinto 2 , S. Solá 1 , R.E. Castro 1 , M.E. Camilo 2 , M.C. Moura 3 , C.M.P. Rodrigues 1 . 1 Centro De Patogénese Molecular, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal; 2 Centro De Nutrição E Metabolismo, IMM, Faculty of Medicine, University of Lisbon, Lisbon, Portugal; 3 Department of Gastroenterology, IMM, Faculty of Medicine, University of Lisbon, Lisbon, Portugal Background/Aims: Apoptosis appears to play a role in the pathogenesis of non-alcoholic (NASH) and alcoholic steatohepatitis (ASH), perhaps contributing to tissue inflammation and fibrogenesis. In this study, we investigated the modulation of apoptosis-related proteins in the liver of patients with steatohepatitis. Methods: Caspase-3 processing was evaluated by immunoblot analysis of liver tissue from 12 patients with NASH, 12 with ASH and 3 histologically normal controls. In addition, expression of death receptors, Bcl-2 family members, and NF-kB inhibitor (IkB) were determined by Western blot. Liver biopsies were also graded for inflammation and fibrosis. Results: Caspase-3 processing was >25% increased in steatohepatitis (p<0.05). Caspase activation coincided with upregulation of proapoptotic Bax (up to 3-fold, p<0.05), confirming the occurrence of apoptosis. Further, production of type I tumor necrosis factor-a receptor was increased 2and 3-fold in NASH and ASH, respectively (p<0.05); the Fas receptor was only slightly elevated. Notably, Bcl-2 expression was strongly enhanced in steatohepatitis (up to 150-fold, p<0.01), but almost absent in control
General Session 3: Molecular Hepatitis
85 ENDOGENOUS OPIOIDS INHIBIT CHOLANGIOCYTE PROLIFERATION IN THE BDL BUT NOT IN THE NORMAL RAT
M. Marzioni 1 , G. Alpini 2 , S. Saccomanno 1 , L. Trozzi 1 , S. Glaser 2 , G. Macarri 1 , A. Benedetti 1 . 1 Dept. of Gastroenterology, Università Delle Marche School of Medicine, Ancona, Italy; 2 The Texas A&M University and Scott&White Hospital HSC, Temple TX, USA In the course of cholestasis the opioid neuromodulation, opioid peptide plasma levels and activity are significantly increased. Proliferating bile ducts express the endogenous opioid precursor Met-enkephalin. Hormoneinduced changes in the Ca2+ signalling reduce cholangiocyte proliferation, interfering with MAP-kinase signalling. Aim: Are endogenous opioids able to regulate cholangiocyte proliferation? Methods: Expression of delta opioid receptor (DOR) in cholangiocytes was studied by immunohistochemistry and immunoblots in pure cholangiocytes from normal and 1 week BDL rats. Pure cholangiocytes from normal and 1 week BDL rats were incubated in vitro with the selective