- Email: [email protected]
CD14 promoter polymorphism associated with risk of NASH
Parallel Session 6: Nutrition and Metabolism weeks) were not significantly different. WHO grade 4 side effects were rare in both arms and treatment-associated mortality was not observed. In addition, there was no difference in Quality of Life in response to treatment. Conclusions: TACO compared to TAC does not improve treatment resuits in advanced HCC and both modalities are equal in terms of toxicity, treatment associated mortality, and quality of life.
established risk factors for NAFLD. Homozygosity for the CD14 T allele was significantly higher in NASH patients compared to FL patients (41% versus 19%; OR: 3.0 [1.1-8.3]), and controls (22%; OR: 2.5 [1.1-5.75]). The TLR4 polymorphism was equally distributed across all groups. This association between a high activity promoter polymorphism in the CD14 receptor and NASH provides strong evidence supporting a role for endotoxin in the pathogenesis of this disease.
[ Parallel Session 6: Nutrition and Metabolism
-~ EGF INCREASES c-Myc, WHICH BLOCKS THE BINDING OF C/EBP TO DNA TO DOWNREGULATE CYP EXPRESSION Marina Tinel I , Alain Berson I , Johny Elkahwaji I , Thierry Cresteil 2, Dominique Pessayre I . IlNSERM U481, Beaujon Hospital, Clichy 92118;
2CNRS UMR 8532, Villejuif,France Cytochromes P450 (CYPs) and other proteins that are transactivated by C/EBPs, are expressed in differentiated hepatocytes, but downregulated in growth-stimulated cells. We determined how EGF dowuregulates CYPs. Rat hepatocytes were cultured with or without various substances for 72 hours, and EGF for the last 48 hours. EGF increased c-myc mRNA and protein, and decreased CYP mRNAs and proteins. Both c-myc overexpression and CYP dowuregulation were prevented by retinoic acid or dimethyl sulfoxide (blocking c-myc transcription), or by two c-myc antisense oligonucleotides (impairing mRNA stability and translation, respectively). A monoclonal anti-c-Myc antibody immunoprecipitated c-Myc and C/EBPalpha and beta from nuclear extracts. Although EGF did not change C/EBPalpha levels and increased C/EBPbeta in nuclear extracts, the EGF-mediated overexpression of c-Myc suppressed the binding of C/EBP dimers to a C/EBP-binding DNA probe. This binding was restored when EGF-treated cells were cotreated with c-myc antisense oligomers (preventing c-Myc induction) or when their nuclear extracts were treated with a polyclonal anti-c-Myc antibody (sequestering c-Myc) prior to the mobility shift assay. Finally, in cells not treated with EGF, CYPs were decreased by an antisense C/EBPalpha oligomer. In conclusion, EGF-mediated c-Myc overexpression blocks the binding of C/EBPs to C/EBP-binding sites on DNA (probably by forming inactive c-Myc-C/EBP complexes) to decrease C/EBPmediated CYP transactivation.
-] CD14 PROMOTER POLYMORPHISM ASSOCIATED WITH RISK OF NASH C. Day. Centre For Liver Research, Newcastle University, UK Non-alcoholic fatty liver disease (NAFLD) covers a spectrum of histological appearances indistinguishable from alcoholic liver disease occurring in none/light drinkers. The first stage - steatosis - is benign, while the second stage - non-alcoholic steatohepatitis (NASH) - progresses to cirrhosis in 20-25% of patients. Steatosis appears to be primarily the consequence of insulin resistance, however, the reasons why only a minority of patients go on to develop NASH are unclear. Studies in animals/humans with NAFLD suggest that gut-derived endotoxin plays a role in NASH by activating macrophages in and outside the liver. Macrophages possess 2 endotoxin receptors CD14 and TLR-4, and functional polymorphisms have been reported in both genes. The CD14 -159 C to T mutation is associated with increased CD14 expression and the Asp299Gly TLR4 mutation with hyporesponsiveness to endotoxin. The aim of this study was to determine whether either of these polymorphisms is associated with NASH. We genotyped 76 patients with biopsy-proven non-alcoholic fatty liver (FL) or NASH (n = 34) and 111 ethnically matched healthy controls for the 2 polymorphisms. FL and NASH patients were similar with respect to the
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HIGH PREVALENCE OF NON SPECIFIC AUTOANTIBODIES (NOSA) IN NAFLD: DOES INSULIN RESISTANCE PLAY A ROLE?
Paola Loria l , Amedeo Lonardo 3, Maria Cristina Fontana I , P. Muratori 2, E Cassani z, Francesca Leonardi l , Dorval Ganazzi 1, Anna Maria Verrone l, Alberto Bagni 4, Andrea Borsatti l, EB. Bianchi 2, Nicola Carulli i. 1Departmentof Internal Medicine, University of
Modena; 2Department of lnternal Medicine, University of Bologna; 3Division of Gastroenteroloy, City Hospital, Modena; 4Department of Pathology, University of Modena, Italy The prevalence of non-organ specific autoantibodies (NOSA) has been reported to be higher in non alcoholic steatohepatitis (NASH) as compared to the general healthy population but no studies have evaluated the prevalence and significance of NOSA in unselected non-alcoholic fatty liver disease (NAFLD). Methods: Serum samples of 84 consecutive subjects with clinical and ultrasonographic evidence of NAFLD were tested for the presence of NOSA by indirect immunoflorescence on frozen sections of rat kidney, liver and stomach and HEp2 cells. Biochemical (ALT, AST, Bilirubin, AE GGT, glucose, fasting insulin, QUICK/, a quantitative test of insulin sensitivity, uric acid, gammaglobulin, indexes of iron metabolism) and anthropometric parameters (BMI, waist/hip ratio) were also evaluated. Results: Two subjects were positive for AMA and were excluded from the analysis. The overall prevalence of NOSA (ANA+SMA) was 34.1% (28/82), 18 subjects (21.4%) being ANA+ and 4 (4.7%) SMA+. Six subjects were positive for both ANA and SMA. NOSA-positive subjects were older (54 + 2 y) than NOSA-negative ones (46 + 2 y) p < 0.01 and mostly females (63.3%). No significant difference was found in the agecorrected mean values of biochemical and morphometric parameters between NOSA-positive and -negative subjects. However, patients with ANA > 1:100 had significantly greater age-corrected mean values of ferritin and insulin than NOSA-negative (p < 0.05). No differences were detected comparing SMA+ and NOSA-patients. Conclusions: The high prevalence of NOSA and the association of higher ANA titers with increased insulin and ferritin values suggest the possibility that insulin resistance in NAFLD might favour the appearance of immunologic phenomena in genetically predisposed individuals.
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HISTOLOGICAL FINDINGS IN THE LIVER BIOPSY OF MORBIDLY OBESE PATIENTS WITH NORMAL LIVER ENZYMES
Ricardo Nathaniel Sepulveda l, Francisco J. Bosques 1, Ricardo Salinas z, Pablo Zorrilla 2, Roberto Rumbaut 3, Hector Maldonado 1, Lucio Vera 4 .
IDepartment of Gastroenterology, UniversidadAutonoma de Nuevo Leon, Monterrey; 2CentroMedico Hidalgo, Monterrey; 3Hospital San Jose, Monterrey; 4Dermatology Laboratory, UniversidadAutonoma de Nuevo Leon, Mexico
Background: Morbid Obesity (MO) is a rising problem in western world countries. Non-alcoholic steatohepatitis (NASH) and liver fibrosis may appear in a subgroup of patients without any clinical or biochemical evidence of liver disease (LD). Objective: We investigated the frequency of histologic changes in the liver biopsy of patients with MO without clinical evidence of LD. Methods: We analyzed the liver biopsies of 40 patients with BMI 47 + 9 and no history of alcoholism or LD, who underwent gastric by-pass
established risk factors for NAFLD. Homozygosity for the CD14 T allele was significantly higher in NASH patients compared to FL patients (41% versus 19%; OR: 3.0 [1.1-8.3]), and controls (22%; OR: 2.5 [1.1-5.75]). The TLR4 polymorphism was equally distributed across all groups. This association between a high activity promoter polymorphism in the CD14 receptor and NASH provides strong evidence supporting a role for endotoxin in the pathogenesis of this disease.
[ Parallel Session 6: Nutrition and Metabolism
-~ EGF INCREASES c-Myc, WHICH BLOCKS THE BINDING OF C/EBP TO DNA TO DOWNREGULATE CYP EXPRESSION Marina Tinel I , Alain Berson I , Johny Elkahwaji I , Thierry Cresteil 2, Dominique Pessayre I . IlNSERM U481, Beaujon Hospital, Clichy 92118;
2CNRS UMR 8532, Villejuif,France Cytochromes P450 (CYPs) and other proteins that are transactivated by C/EBPs, are expressed in differentiated hepatocytes, but downregulated in growth-stimulated cells. We determined how EGF dowuregulates CYPs. Rat hepatocytes were cultured with or without various substances for 72 hours, and EGF for the last 48 hours. EGF increased c-myc mRNA and protein, and decreased CYP mRNAs and proteins. Both c-myc overexpression and CYP dowuregulation were prevented by retinoic acid or dimethyl sulfoxide (blocking c-myc transcription), or by two c-myc antisense oligonucleotides (impairing mRNA stability and translation, respectively). A monoclonal anti-c-Myc antibody immunoprecipitated c-Myc and C/EBPalpha and beta from nuclear extracts. Although EGF did not change C/EBPalpha levels and increased C/EBPbeta in nuclear extracts, the EGF-mediated overexpression of c-Myc suppressed the binding of C/EBP dimers to a C/EBP-binding DNA probe. This binding was restored when EGF-treated cells were cotreated with c-myc antisense oligomers (preventing c-Myc induction) or when their nuclear extracts were treated with a polyclonal anti-c-Myc antibody (sequestering c-Myc) prior to the mobility shift assay. Finally, in cells not treated with EGF, CYPs were decreased by an antisense C/EBPalpha oligomer. In conclusion, EGF-mediated c-Myc overexpression blocks the binding of C/EBPs to C/EBP-binding sites on DNA (probably by forming inactive c-Myc-C/EBP complexes) to decrease C/EBPmediated CYP transactivation.
-] CD14 PROMOTER POLYMORPHISM ASSOCIATED WITH RISK OF NASH C. Day. Centre For Liver Research, Newcastle University, UK Non-alcoholic fatty liver disease (NAFLD) covers a spectrum of histological appearances indistinguishable from alcoholic liver disease occurring in none/light drinkers. The first stage - steatosis - is benign, while the second stage - non-alcoholic steatohepatitis (NASH) - progresses to cirrhosis in 20-25% of patients. Steatosis appears to be primarily the consequence of insulin resistance, however, the reasons why only a minority of patients go on to develop NASH are unclear. Studies in animals/humans with NAFLD suggest that gut-derived endotoxin plays a role in NASH by activating macrophages in and outside the liver. Macrophages possess 2 endotoxin receptors CD14 and TLR-4, and functional polymorphisms have been reported in both genes. The CD14 -159 C to T mutation is associated with increased CD14 expression and the Asp299Gly TLR4 mutation with hyporesponsiveness to endotoxin. The aim of this study was to determine whether either of these polymorphisms is associated with NASH. We genotyped 76 patients with biopsy-proven non-alcoholic fatty liver (FL) or NASH (n = 34) and 111 ethnically matched healthy controls for the 2 polymorphisms. FL and NASH patients were similar with respect to the
21
•
-
]
HIGH PREVALENCE OF NON SPECIFIC AUTOANTIBODIES (NOSA) IN NAFLD: DOES INSULIN RESISTANCE PLAY A ROLE?
Paola Loria l , Amedeo Lonardo 3, Maria Cristina Fontana I , P. Muratori 2, E Cassani z, Francesca Leonardi l , Dorval Ganazzi 1, Anna Maria Verrone l, Alberto Bagni 4, Andrea Borsatti l, EB. Bianchi 2, Nicola Carulli i. 1Departmentof Internal Medicine, University of
Modena; 2Department of lnternal Medicine, University of Bologna; 3Division of Gastroenteroloy, City Hospital, Modena; 4Department of Pathology, University of Modena, Italy The prevalence of non-organ specific autoantibodies (NOSA) has been reported to be higher in non alcoholic steatohepatitis (NASH) as compared to the general healthy population but no studies have evaluated the prevalence and significance of NOSA in unselected non-alcoholic fatty liver disease (NAFLD). Methods: Serum samples of 84 consecutive subjects with clinical and ultrasonographic evidence of NAFLD were tested for the presence of NOSA by indirect immunoflorescence on frozen sections of rat kidney, liver and stomach and HEp2 cells. Biochemical (ALT, AST, Bilirubin, AE GGT, glucose, fasting insulin, QUICK/, a quantitative test of insulin sensitivity, uric acid, gammaglobulin, indexes of iron metabolism) and anthropometric parameters (BMI, waist/hip ratio) were also evaluated. Results: Two subjects were positive for AMA and were excluded from the analysis. The overall prevalence of NOSA (ANA+SMA) was 34.1% (28/82), 18 subjects (21.4%) being ANA+ and 4 (4.7%) SMA+. Six subjects were positive for both ANA and SMA. NOSA-positive subjects were older (54 + 2 y) than NOSA-negative ones (46 + 2 y) p < 0.01 and mostly females (63.3%). No significant difference was found in the agecorrected mean values of biochemical and morphometric parameters between NOSA-positive and -negative subjects. However, patients with ANA > 1:100 had significantly greater age-corrected mean values of ferritin and insulin than NOSA-negative (p < 0.05). No differences were detected comparing SMA+ and NOSA-patients. Conclusions: The high prevalence of NOSA and the association of higher ANA titers with increased insulin and ferritin values suggest the possibility that insulin resistance in NAFLD might favour the appearance of immunologic phenomena in genetically predisposed individuals.
•
HISTOLOGICAL FINDINGS IN THE LIVER BIOPSY OF MORBIDLY OBESE PATIENTS WITH NORMAL LIVER ENZYMES
Ricardo Nathaniel Sepulveda l, Francisco J. Bosques 1, Ricardo Salinas z, Pablo Zorrilla 2, Roberto Rumbaut 3, Hector Maldonado 1, Lucio Vera 4 .
IDepartment of Gastroenterology, UniversidadAutonoma de Nuevo Leon, Monterrey; 2CentroMedico Hidalgo, Monterrey; 3Hospital San Jose, Monterrey; 4Dermatology Laboratory, UniversidadAutonoma de Nuevo Leon, Mexico
Background: Morbid Obesity (MO) is a rising problem in western world countries. Non-alcoholic steatohepatitis (NASH) and liver fibrosis may appear in a subgroup of patients without any clinical or biochemical evidence of liver disease (LD). Objective: We investigated the frequency of histologic changes in the liver biopsy of patients with MO without clinical evidence of LD. Methods: We analyzed the liver biopsies of 40 patients with BMI 47 + 9 and no history of alcoholism or LD, who underwent gastric by-pass