M. Carbone

Correlation Between Liver Fibrosis and Inflammation in Patients Transplanted for HCV Liver Disease

Hepatitis C virus (HCV) re‐infection after liver transplantation (LT) is characterized by an accelerated disease progression in recent years with unclear mechanisms. We evaluate the relationship between progression of liver fibrosis and histological necro‐inflammation in HCV recipients, according to age of transplant. Fifty‐five patients transplanted (1993–2002) for HCV liver disease, were included in the study. Recipients were retrospectively stratified in three different age of transplant, of 40 months each: group 1) from January 1993 to May 1996; group 2) from June 1996 to august 1999; group 3) from September 1999 to December 2002. Grading (necro‐inflammation) and staging (fibrosis) scores were evaluated in liver biopsies at 1, 2 and 3 years from LT (Ishak classification). For all age of transplant the main factor associated with fibrosis progression, was grading score (p < 0.05). However mean staging score for each point of grading increased from 0.3 ± 0.2 in older LT to 0.7 ± 0.5 in newer ones (p = 0.01). In conclusion in HCV–LT patients (1) liver fibrosis is strictly associated to histological necro‐inflammation; (2) the proportion of this relationship has been changing in recent years since newer LT patients, show an increased amount of fibrosis in comparison with the older ones, for similar grading score.

TUDCA prevents cholestasis and canalicular damage induced by ischemia‐reperfusion injury in the rat, modulating PKCα–ezrin pathway

Cholestasis, induced by liver ischemia‐reperfusion injury (IRI), is characterized by dilatation of bile canaliculi and loss of microvilli. Tauroursodeoxycholic acid (TUDCA) is an anti‐cholestatic agent, modulating protein kinase C (PKC) α pathway. PKC reduces ischemic damage in several organs, its isoform α modulates ezrin, a key protein in the maintenance of cell lamellipoidal extensions. We evaluated the effects of TUDCA on cholestasis, canalicular changes and PKCα–ezrin expression in a rat model of liver IRI. Livers flushed and stored with Belzer solution or Belzer + 10 mm TUDCA (4 °C for 6 h) were reperfused (37 °C with O2) with Krebs–Ringer bicarbonate + 2.5 μmol/min of Taurocholate or TUDCA. Bile was harvested for bile flow assessment. Liver tissue was employed for Electron Microscopy (EM) and for PKCα and ezrin immunoblot and immunofluorescence. The same experiments were conducted with the PKCα inhibitor Go‐6976. TUDCA‐treated livers showed increased bile flow (0.25 ± 0.17 vs. 0.042 ± 0.02 μl/min/g liver, P < 0.05) and better preservation of microvilli and bile canalicular area at EM. These effects were associated with increased PKCα and ezrin expression (P = 0.03 and P = 0.04 vs. control respectively), as also confirmed by immunofluorescence data. PKCα inhibition abolished these TUDCA effects. TUDCA administration during IRI reduces cholestasis and canalicular damage in the liver modulating PKCα–ezrin pathway.

Leptin attenuates ischemia–reperfusion injury in the rat liver

Leptin is an adipocytokine that reduces ischemic damage in several organs including brain and heart. STAT3 activation is a key step for the attainment of leptin effects in various tissues. We evaluated the possible effect of leptin on liver viability and STAT3 activation, in a rat model of ischemia–reperfusion injury. Rat livers, flushed and stored with Belzer solution (4° C for 24 h), were warmly reperfused (3.5 ml/min/g liver for 1 h at 37° C with O2) with Krebs–Ringer bicarbonate. Treatment group underwent an identical protocol with the adjunct of Leptin (10 ng/ml). Liver effluent was harvested to assess LDH and AST output. Liver tissue was used for pSTAT3 expression (western blot and immunostaining), optical microscopy, TUNEL, and Cell Death Detection assays. The pSTAT3 expression was enhanced by administration of leptin. In parallel, LDH and AST output were reduced (P = 0.04 and P = 0.02 for LDH and AST, respectively). Optical microscopy, TUNEL, and Cell Death Detection assay results demonstrated increased viability in livers treated with leptin in comparison with others (Optical microscopy P = 0.02; TUNEL P = 0.01; Cell death Detection assay P = 0.003). In conclusion, cold storage and reperfusion with leptin reduce liver ischemia–reperfusion injury. This effect is associated with an increased expression of pSTAT‐3.

597 SERUM/ERITROCYTE RIBAVIRIN X 100 RATIO AS AN INDICATOR OF SVR IN HCV GENOTYPE-1 PATIENTS OBTAINING EVR

ackground. Virologic predictors during treatment with eg-interferon and ribavirin of hepatitis C are useful as hey allow early discontinuation of an unnecessary and xpensive treatment in non-responders. In this setting early irologic response (EVR), defined as viral load decline 2 log10 or undetectable HCV-RNA at week 12 of treatents has emerged as a useful tool to continue treatment. nfortunately, while EVR negative predictive value (NPV) s high, as approximately 2% only of patients not matchng EVR may achieve a sustained virological response SVR), the positive predictive value (PPV), in particular in enotype-1 patients, is low (∼60%). Other indicators may be eeded to predict SVR and in particular in those obtaining VR. im. To evaluate the usefulness of serum and eritrocyte ribvirin levels in predicting SVR in HCV genotype-1 patients ndergoing peginterferon + ribavirin treatment. ethods. 30 HCV genotype-1 patients (22 M/8F mean age 5.2± 13.6 years) undergoing a standard treatment schedle (Peg-IFN 180 mcg weekly + ribavirin 1000 or 1200 mg aily, according to body weight) were included in the study. lasma and eritrocyte ribavirin levels were evaluated in all atients at week 12 and at week 24 in those obtaining EVR nly. Ribavirin concentration was evaluated by high perforance liquid chromatography after solid phase extraction and mploying 3-methyl-cytidine as internal standard. Patients iochemical data were also recorded. esults. There was no difference among EVR and nonVR patients in terms of serum and eritrocyte ribavirin oncentration at week 12. At week 24, EVR patients obtainng SVR exhibited higher levels of ribavirin in serum nd lower in eritrocytes, in comparison with non-SVR atients (serum 14.1± 10.5 M vs. 5.9± 4.1 M; p< 0.02; ritrocyte 1072± 420 M vs. 1793± 903 M; p< 0.02). hen ([serum ribavirin]/[eritrocyte ribavirin]× 100) ratio as compared among these two groups, the difference was nhanced (1.6± 1.5 vs. 0.4± 0.34; p< 0.01). Receiver operting characteristic (ROC) curve analysis identified a cut-off or ([serum ribavirin]/[eritrocyte ribavirin]× 100) ratio in redicting SVR of 0.6, with a NPV of 80% and a PPV f 85%, while those related to EVR were 100% and 63%, espectively. onclusion. ([serum ribavirin]/[eritrocyte ribavirin]× 100) atio at week 24 seems to be an 85% indicator of SVR in a I p n isease 41 (2009) A1–A45 A35