POSTERS

(NGAL) is one of the most up-regulated genes. NGAL gene

expression was assessed in patients with AH and other types liver

diseases. NGAL expression was also studied in animal models of

acute and chronic liver injury (CCl4), acute-on-chronic liver injury

(CCl4 plus LPS) and alcoholic liver disease (CCl4 plus ethanol) is

a cytokine typically released by activated neutrophils. Finally, the

effects of recombinant human NGAL was assed in hepatic stellate

cells (HSC).

Results: NGAL gene expression was markedly up-regulated in

patients with AH compared to other liver disease such as HCV,

compensated alcoholic cirrhosis and NASH. Serum levels were

also found elevated in patients with AH. Importantly, the baseline

hepatic mRNA expression of NGAL correlated with disease severity

and 90-day survival. Hepatic NGAL expression was induced by an

acute and acute-on-chronic liver injury and in mice with alcoholic

liver disease. Finally recombinan NGAL induced procollagen I and

MCP-1 expression in human HSC.

Conclusions: NGAL is markedly overexpressed in AH and in animal

models of acute-on-chronic liver injury and it exerts inflammatory

effects in hepatic cells. These results suggest that NGAL is a potential

target for therapy in AH.

12. AUTOIMMUNE AND CHRONICCHOLESTATIC LIVER DISEASE

P345

PRIMARY PREVENTION OF GALLBLADDER STONES DURING

WEIGHT LOSS: SYSTEMATIC REVIEW AND META-ANALYSIS OF

RANDOMISED CONTROLLED TRIALS

C.S. Stokes1, L.L. Gluud2, M. Casper1, F. Lammert1. 1Department of

Medicine II, Saarland University Medical Center, Homburg, Germany;2Department of Internal Medicine, University of Copenhagen, Hellerup,

Denmark

E-mail: caroline.stokes@uks.eu

Background and Aims: The incidence of gallstones is rising

partly due to the obesity epidemic. Cholecystectomy for

(symptomatic) gallbladder stones poses a major burden on

healthcare resources. We systematically evaluated the efficacy of

non-surgical interventions for the primary prevention of gallbladder

stones in obese adults undergoing weight loss, which is a known

risk factor for stone formation.

Methods: A Cochrane review of randomised controlled trials

comparing ursodeoxycholic acid (UDCA) or a high-fat weight loss

diet versus control interventions was conducted. Electronic and

manual searches identified a total of 13 trials comprising 1,837

obese participants undergoing weight loss through dieting or after

bariatric surgery. Random-effects meta-analyses were performed

to assess the efficacy of these interventions with regards gallstone

prevention. Inter-trial heterogeneity and bias were evaluated with

subgroup, sensitivity, regression, and sequential analyses.

Results: UDCA reduced the risk of ultrasonically verified gallstones

compared with control interventions (RR 0.33, 95%CI 0.18 to

0.60, I2=65%, NNT=9). UDCA was more beneficial in participants

undergoing weight loss through diet alone (RR 0.17, 95%CI 0.11 to

0.25, I2=0%) than after bariatric surgery (RR 0.42, 95%CI 0.21 to

0.83, I2=64%). A high versus a low dietary fat content also reduced

gallstones (RR 0.09, 95%CI 0.01 to 0.61, I2=0%). No major adverse

events were reported, and regression analysis showed no evidence

of small study effects.

Conclusions: The results of this meta-analysis indicate that during

weight loss, UDCA and a high-fat weight loss diet prevent formation

of gallstones and show potential as cost-effective alternatives to

cholecystectomy.

P346

TAUROURSODESOXICOLATE (TUDC) PREVENTS ACTIVATION

OF THE PRO-CHOLESTATIC SIGNALLING PATHWAYS, cPKC AND

PI3K/Akt, IN ESTRADIOL 17b-D-GLUCURONIDE (E217G)-INDUCED

CHOLESTASIS

A.C. Boaglio, G.S. Miszczuk, I.R. Barroso, F.D. Toledo, F.A. Crocenzi,

M.G. Roma. National University of Rosario, Institute of Experimental

Physiology (IFISE) – CONICET, Rosario, Argentina

E-mail: mroma@fbioyf.unr.edu.ar

Background and Aims: E217G induces cholestasis by triggering

endocytosis and further intracellular retention of Bsep and Mrp2,

in a cPKC- and PI3K-dependent manner, respectively. Pregnancy

induced cholestasis, associated with high E217G levels, is routinely

treated with ursodeoxycholic acid (UDCA). Since protective UDCA

action mechanisms in E217G-induced cholestasis have not been

elucidated, we ascertained here whether TUDC, the main, active

UDCA metabolite, prevents activation of cPKC and PI3K/Akt.

Methods: Activation of cPKC and PI3K/Akt was evaluated in isolated

rat hepatocytes by immunoblotting (assessment of membrane-

bound and phosphorylated form, respectively). Bsep/Mrp2 function

was quantified in isolated rat hepatocyte couplets (IRHCs)

by assessing canalicular vacuolar accumulation (CVA) of their

fluorescent substrates, CGamF and GS-MF, respectively. We also

studied the preventive mechanisms of TUDC in E217G-induced

cholestasis in isolated, perfused rat liver (IPRL), and Bsep/Mrp2

localization by immunofluorescence.

Results: E217G (200mM) activated both cPKC and PI3K/Akt.

Pretreatment with TUDC (100mM) prevented activation of cPKC

(−34±4%) and PI3K/Akt (−37±2%); p < 0.05 vs. E217G alone. The

E217G-mediated decrease in CVA of cGamF (−59±2%) and GS-

MF (−55±3%) was partially prevented by TUDC (+72±6%) and

(+78±3%); p < 0.05 vs. E217G alone. In IPRL, E217G (2mmol/liver)

induced an acute decrease in bile flow (−59±14%; p < 0.001),

which was completely prevented by TUDC (p < 0.001). A

similar behaviour was observed for the biliary excretion of

the Bsep and Mrp2 substrates, TC and DNP-SG, respectively.

Immunofluorescence revealed that TUDC prevented E217G-induced

Bsep/Mrp2 endocytosis.

Conclusions: TUDC restores function and localization of Bsep/Mrp2

as impaired by E217G, by preventing both cPKC and PI3K/Akt

activation.

P347

INCIDENCE AND RISK FACTORS FOR EXTRA-HEPATIC

MALIGNANCIES (EM) IN PRIMARY BILIARY CIRRHOSIS:

A COMPARATIVE STUDY FROM TWO EUROPEAN REFERRAL

CENTERS

N. Cazzagon1, A. Pares2, A. Spinazze1, L. Caballeria2, A. Reig2,

I. Franceschet1, V. Baldo3, A. Buja3, P. Furlan3, A. Floreani1.1Department of Surgery, Oncology and Gastroenterology, University of

Padua, Padua, Italy; 2Liver Unit, Digestive Diseases Institute, Centro

de Investigacion Biomedica en Red de Enfermedades Hepaticas y

Digestivas, Hospital Clınic, IDIBAPS, Barcelona, Spain; 3Department

of Molecular Medicine, Public Health Section, University of Padua,

Padua, Italy

E-mail: nora.cazzagon@gmail.com

Background and Aims: There is limited information on the

prevalence/incidence, survival, and risk factors for developing EM

in PBC.

Aim: To analyze the incidence/prevalence, risk factors and survival

for EM in patients with PBC in two European centers.

Methods: The study was carried out in two series of PBC patients

(361 of Padova, Italy and 397 of Barcelona, Spain) followed-up

for a mean period of 7.7±7 and 12.2±7 years respectively. The

incidence of EM was compared to the estimated incidence data

from IARC. Demographic features and factors associated with tumor

S182 Journal of Hepatology 2014 vol. 60 | S67–S214