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The role of serum leptin level in progression from steatosis to steatohepatitis
Category 8: Nutrition, metabolism, alcoholic liver disease, pharmacology sinusoidal fibrosis were more marked, whereas grade of inflammation was less intense. HCV genoptype 3 was associated with higher steatosis score. Conclusions: 1. NASH lesions can be found in patients with CrHC, particularly in obese, hyperlipemic women. These patients show significantly higher scores of inflammation and fibrosis; 2. HCV genotype 3 is associated with higher steatosis scores.
~ - " ~ ROLE OF MUTATIONS OF HFE GENE IN NON-ALCOHOLIC STEATOHEPATITIS AND ALCOHOLIC LIVER DISEASE Tereza Pucelikova l , Blanka Cieslarova 1, Ivana Putova 1, Iva Hruba 2, Jan Stritesky 3, Vladimir Palicka 4, Jiri Horak I . llst Departement of
Medicine, 3rd Medical Faculty, Charles University, Prague; 2Departement of Medicine, Medical Faculty, Charles University, Hradec Kralove; 3Departement of Pathology, 1st Medical Faculty, Charles University, Prague; 4Departement of Clinical Chemistry, Medical Faculty, Charles University, Hradec Kralove, Czech Republic Serum iron indices and hepatic iron are known to be elevated in nonalcoholic steatohepatitis (NASH) and alcoholic liver disease. C282Y mutation in HFE gene was identified to cause primary hemochromatosis in homozygotes. H63D mutation in the same gene may contribute to iron overload in compound heterozygotes. Aim: To determine whether there is an association between parametres of iron metabolism and mutations in HFE gene in NASH and alcoholic liver disease. Patients and Methods: Three groups of patients were included: patients with NASH (n = 28), alcoholic stetofibrosis (n = 29) and alcoholic cirrhosis (n -- 31). All subjects were screened for two major HFE mutations C282Y a H63D by RFLE HFE genotypes were compared with indices of iron metabolism (serum iron, ferritin, transferrin saturation) and hepatic iron contents (HIC). HICs were measured by atom-absorption spectrophotometry. Results: C282Y allele frequency was 6.9% in the group of NASH, 0% in non-alcoholic steatohepatitis and 5.4% for patients with alcoholic liver disease, H63D allele frequencies were 25%, 22% and 14% for the respective groups. Alelle frequencies of C282Y and H63D in healthy Czech population are 4.6% and 22%, respectively. Conclusions: We found higher prevalence of H63D and C282Y mutations in NASH patients in comparison to to healthy population, however this difference was not statistically significant. No differences were observed for the patients with alcoholic liver disease. None of tested mutations seems to contribute to elevation of ferritin, transferrin saturation and hepatic iron content in patients with NASH and alcoholics.
~-5"I FKS06 AND RAPAMYCIN REDUCE NITRIC OXIDE PRODUCTION AND NUCLEAR FACTOR KAPPA B ACTIVATION IN CULTURED HEPATOCYTES S. Sanchez-Campos l , B. Gutierrez l , J.M. Culebras 2, J. Gonzalez-Gallego i M.J. Tunon 1. lDepartment of Physiology;
university of Ledn, leon; 2of Leon, Leon, Spain FK506 and rapamycin are potent immunosuppressive drugs that inhibit proinflammatory cytokine expression and reduce the citotoxic response in different cell types. The increase in nitric oxide (NO) production following transplantation is associated to acute allograft rejection. Aims: To compare the effect of both drugs on NO production, on inducible nitric oxide synthase (iNOS) expression and on nuclear factor kappa B (NF-kB) activation in lipopolysaccharide (LPS)-treated hepatocytes. Methods: Hepatocytes were obtained from rats receiving LPS (5 mg/kg i.p.) and were isolated by double coUagenase perfusion. Cells were incubated in 5% CO2 with LPS (10 mg/ml medium), LPS + FK506 (10 mM) o LPS + rapamycin (10 mM). Results: LPS increased the release of LDH and nitrite ions, oxidation products of NO, into the culture medium. Simultaneous addition of FK506
151
or rapamycin resulted in an inhibition of LDH and nitrite ions release. Both immunosuppressive drugs inhibited the induction of iNOS mRNA (RT-PCR) and protein (Western blot) stimulated by LPS, although to a greater extent in the case of rapamycin. LPS-induced NF-kB activation was absent in cells treated with FK-506 or rapamycin. Conclusions: Data obtained indicate that FK506 and rapamycin block NFkB activation and inhibit NO production by hepatocytes, confirming the antiinflammatory potential of both drugs and its capacity to reduce acute allograft rejection.
~ - ~ T H E ROLE OF SERUM LEPTIN LEVEL IN PROGRESSION FROM STEATOSIS TO STEATOHEPATITIS Ender Serin, Birol Ozer, Yuksel Gumurdulu, Sedat Boyacioglu.
Gastroenterology, Baskent UniversityAdana Hospital, Adana, Turkey Background: The peripheral insulin resistance is a feature of nonalcoholic steatohepatitis (NASH). In hepatocytes, leptin has complex effects on insulin response and been suggested to increase in patients with NASH. To our knowledge the comparision of serum leptin level in patients with steatosis only and in those with NASH has not been investigated. Our aim was to determine the role of leptin in progression from steatosis to steatohepatitis by comparing the serum levels of this hormon in these groups. Methods: We evaluated 31 patients with steatosis (M/F: 10/21), 32 patients with steatohepatitis (M/F: 18/14), and 8 nonobes and nonsteatotic controls (M/F: 4/4) for body mass index (BMI), insulin resistance (IR), glucose intolerance, diabetes mellitus, and serum levels of lipids and leptin. We stratified each patient groups in two subgroup according to BMI (<30 and >31): A) NASH and <30; B) steatosis and <30; C) NASH and >31; D) steatosis and >31. Results: Age and serum lipids were comparable among all groups. Serum leptin level was higher in D than C (p = 0.017). There was a trend for higher leptin value for B than A (p = 0.097). Logistic regression anaysis showed positive effects of sex and BMI (p < 0.001), and no effect of transaminase elevation and steatosis on leptin level. IR was comparable among four patient groups, but controls had lower IR compared with B. Conclusion: Our data indicate that elevated serum leptin level is not responsible for progression from steatosis to steatohepatitis, instead leptin may decrease due to chronic inflammation during steatohepatitis.
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EFFICACY OF PHLEBOTOMY THERAPHY AND RESTRICTION CALORIC INTAKE IN PATIENTS WITH INSULIN RESISTENCE-ASSOCIATED HEPATIC IRON OVERLOAD SYNDROME (IR-HIO)
Paola Trombini l, Anna Vergani 1, Alessandra Salvioni 1, Mauro Vigano 1, Raffaella Mariani 1, Chiara Corengia 1, Adele Zoppo 2, Giuseppe Boari 3, Alberto Piperno t . l Clinica Medica UniversitaMilano Bicocca Ospedale
S Gerardo Monza; 2Multimedica Sesto S Giovanni Milano; 31stitutodi Statistica Universita Cattolica S Cuore Milano, Italy Background: IR-HIO is a clinical condition characterized by moderate iron overload and metabolic disorders. It is unknown whether iron overload progresses over time and whether iron removal and weight loss have beneficial effects in these patients. Aim: To evaluate the behaviour of metabolic and iron indices in 59 IR-HIO during a no-treatment-follow-up and the effects of phlebotomy therapy and caloric diet. Subjects and Methods: 24 patients were followed-up for at least 24 months; 26 patients underwent phlebotomy therapy; 17 underwent restriction caloric intake. Diagnosis was based on classical criteria. Data were compared by t-test, variance (ANOVA) and covariance analyses. Results and Conclusions: Iron and metabolic indices showed no significant variations during the follow-up period suggesting that iron accumulation does not progress over time in IR-HIO. Serum ALT (p = 0.01), gammaGT (p < 0.01) and triglyceride (p = 0.01) significantly decreased after iron
~ - " ~ ROLE OF MUTATIONS OF HFE GENE IN NON-ALCOHOLIC STEATOHEPATITIS AND ALCOHOLIC LIVER DISEASE Tereza Pucelikova l , Blanka Cieslarova 1, Ivana Putova 1, Iva Hruba 2, Jan Stritesky 3, Vladimir Palicka 4, Jiri Horak I . llst Departement of
Medicine, 3rd Medical Faculty, Charles University, Prague; 2Departement of Medicine, Medical Faculty, Charles University, Hradec Kralove; 3Departement of Pathology, 1st Medical Faculty, Charles University, Prague; 4Departement of Clinical Chemistry, Medical Faculty, Charles University, Hradec Kralove, Czech Republic Serum iron indices and hepatic iron are known to be elevated in nonalcoholic steatohepatitis (NASH) and alcoholic liver disease. C282Y mutation in HFE gene was identified to cause primary hemochromatosis in homozygotes. H63D mutation in the same gene may contribute to iron overload in compound heterozygotes. Aim: To determine whether there is an association between parametres of iron metabolism and mutations in HFE gene in NASH and alcoholic liver disease. Patients and Methods: Three groups of patients were included: patients with NASH (n = 28), alcoholic stetofibrosis (n = 29) and alcoholic cirrhosis (n -- 31). All subjects were screened for two major HFE mutations C282Y a H63D by RFLE HFE genotypes were compared with indices of iron metabolism (serum iron, ferritin, transferrin saturation) and hepatic iron contents (HIC). HICs were measured by atom-absorption spectrophotometry. Results: C282Y allele frequency was 6.9% in the group of NASH, 0% in non-alcoholic steatohepatitis and 5.4% for patients with alcoholic liver disease, H63D allele frequencies were 25%, 22% and 14% for the respective groups. Alelle frequencies of C282Y and H63D in healthy Czech population are 4.6% and 22%, respectively. Conclusions: We found higher prevalence of H63D and C282Y mutations in NASH patients in comparison to to healthy population, however this difference was not statistically significant. No differences were observed for the patients with alcoholic liver disease. None of tested mutations seems to contribute to elevation of ferritin, transferrin saturation and hepatic iron content in patients with NASH and alcoholics.
~-5"I FKS06 AND RAPAMYCIN REDUCE NITRIC OXIDE PRODUCTION AND NUCLEAR FACTOR KAPPA B ACTIVATION IN CULTURED HEPATOCYTES S. Sanchez-Campos l , B. Gutierrez l , J.M. Culebras 2, J. Gonzalez-Gallego i M.J. Tunon 1. lDepartment of Physiology;
university of Ledn, leon; 2of Leon, Leon, Spain FK506 and rapamycin are potent immunosuppressive drugs that inhibit proinflammatory cytokine expression and reduce the citotoxic response in different cell types. The increase in nitric oxide (NO) production following transplantation is associated to acute allograft rejection. Aims: To compare the effect of both drugs on NO production, on inducible nitric oxide synthase (iNOS) expression and on nuclear factor kappa B (NF-kB) activation in lipopolysaccharide (LPS)-treated hepatocytes. Methods: Hepatocytes were obtained from rats receiving LPS (5 mg/kg i.p.) and were isolated by double coUagenase perfusion. Cells were incubated in 5% CO2 with LPS (10 mg/ml medium), LPS + FK506 (10 mM) o LPS + rapamycin (10 mM). Results: LPS increased the release of LDH and nitrite ions, oxidation products of NO, into the culture medium. Simultaneous addition of FK506
151
or rapamycin resulted in an inhibition of LDH and nitrite ions release. Both immunosuppressive drugs inhibited the induction of iNOS mRNA (RT-PCR) and protein (Western blot) stimulated by LPS, although to a greater extent in the case of rapamycin. LPS-induced NF-kB activation was absent in cells treated with FK-506 or rapamycin. Conclusions: Data obtained indicate that FK506 and rapamycin block NFkB activation and inhibit NO production by hepatocytes, confirming the antiinflammatory potential of both drugs and its capacity to reduce acute allograft rejection.
~ - ~ T H E ROLE OF SERUM LEPTIN LEVEL IN PROGRESSION FROM STEATOSIS TO STEATOHEPATITIS Ender Serin, Birol Ozer, Yuksel Gumurdulu, Sedat Boyacioglu.
Gastroenterology, Baskent UniversityAdana Hospital, Adana, Turkey Background: The peripheral insulin resistance is a feature of nonalcoholic steatohepatitis (NASH). In hepatocytes, leptin has complex effects on insulin response and been suggested to increase in patients with NASH. To our knowledge the comparision of serum leptin level in patients with steatosis only and in those with NASH has not been investigated. Our aim was to determine the role of leptin in progression from steatosis to steatohepatitis by comparing the serum levels of this hormon in these groups. Methods: We evaluated 31 patients with steatosis (M/F: 10/21), 32 patients with steatohepatitis (M/F: 18/14), and 8 nonobes and nonsteatotic controls (M/F: 4/4) for body mass index (BMI), insulin resistance (IR), glucose intolerance, diabetes mellitus, and serum levels of lipids and leptin. We stratified each patient groups in two subgroup according to BMI (<30 and >31): A) NASH and <30; B) steatosis and <30; C) NASH and >31; D) steatosis and >31. Results: Age and serum lipids were comparable among all groups. Serum leptin level was higher in D than C (p = 0.017). There was a trend for higher leptin value for B than A (p = 0.097). Logistic regression anaysis showed positive effects of sex and BMI (p < 0.001), and no effect of transaminase elevation and steatosis on leptin level. IR was comparable among four patient groups, but controls had lower IR compared with B. Conclusion: Our data indicate that elevated serum leptin level is not responsible for progression from steatosis to steatohepatitis, instead leptin may decrease due to chronic inflammation during steatohepatitis.
~-~
EFFICACY OF PHLEBOTOMY THERAPHY AND RESTRICTION CALORIC INTAKE IN PATIENTS WITH INSULIN RESISTENCE-ASSOCIATED HEPATIC IRON OVERLOAD SYNDROME (IR-HIO)
Paola Trombini l, Anna Vergani 1, Alessandra Salvioni 1, Mauro Vigano 1, Raffaella Mariani 1, Chiara Corengia 1, Adele Zoppo 2, Giuseppe Boari 3, Alberto Piperno t . l Clinica Medica UniversitaMilano Bicocca Ospedale
S Gerardo Monza; 2Multimedica Sesto S Giovanni Milano; 31stitutodi Statistica Universita Cattolica S Cuore Milano, Italy Background: IR-HIO is a clinical condition characterized by moderate iron overload and metabolic disorders. It is unknown whether iron overload progresses over time and whether iron removal and weight loss have beneficial effects in these patients. Aim: To evaluate the behaviour of metabolic and iron indices in 59 IR-HIO during a no-treatment-follow-up and the effects of phlebotomy therapy and caloric diet. Subjects and Methods: 24 patients were followed-up for at least 24 months; 26 patients underwent phlebotomy therapy; 17 underwent restriction caloric intake. Diagnosis was based on classical criteria. Data were compared by t-test, variance (ANOVA) and covariance analyses. Results and Conclusions: Iron and metabolic indices showed no significant variations during the follow-up period suggesting that iron accumulation does not progress over time in IR-HIO. Serum ALT (p = 0.01), gammaGT (p < 0.01) and triglyceride (p = 0.01) significantly decreased after iron