studies

616A AASLD ABSTRACTS HEPATOLOGY, October, 2015
Disclosures:
The following authors have nothing to disclose: Ying Wan, Shannon S. Glaser,
Nan Wu, Fanyin Meng, Julie Venter, Romina Mancinelli, Heather L. Francis,
Antonio Franchitto, Paolo Onori, Tina Kyritsi, Shanika Avila, Guido Carpino,
Holly A. Standeford, Gianfranco Alpini, Eugenio Gaudio
818
Pre-treatment with TIMP-3 prevents the development of
biliary injury in an LPS enhanced ischaemia-reperfusion
animal model
Janske Reiling 1,2 , Kim Bridle 1,3 , Catherine M. Campbell 4 , Nishreen
Santrampurwala 1,3 , Laurence J. Britton 1,3 , Ari J. Cohen 5 , Darrell H.
Crawford 1,3 , Cornelis H. Dejong 2,6 , Jonathan Fawcett 1,7 ; 1 school
of medicine, University of Queensland, Brisbane, QLD, Australia;
2 NUTRIM School of Nutrition and Translational Research in Metabolism,
Maastricht University, Maastricht, Netherlands; 3 Gallipoli
Medical Reserach Foundation, Brisbane, QLD, Australia; 4 Envoi
Specialist Pathologists, Brisbane, QLD, Australia; 5 Laboratory of
Transplant Reseach, Institute of Translational Research, Ochsner
Clinical Foundation, New Orleans, LA; 6 Department of Surgery,
Maastricht University Medical Centre, Maastricht, Netherlands;
7 Queensland Liver Transplant Service, Princess Alexandra Hospital,
Brisbane, QLD, Australia
Introduction Transplantation of donor livers retrieved after circulatory
death is often complicated by the development of biliary
injury and subsequent stricture formation. Our recent studies
identified a role for lipopolysaccharides (LPS) in the development
of biliary injury. This study aimed to assess whether
inhibition of TNF-α cleavage into its biologically active form,
by pre-treatment with tissue inhibitor of metalloproteinases-3
(TIMP-3), would prevent the development of biliary injury.
Methods Male Sprague Dawley rats underwent either sham
surgery (sham), 1000ng/kg human recombinant TIMP-3 one
hour prior to sham surgery (TIMP-3), 70% partial liver ischaemia
for thirty minutes with 1mg/kg LPS (combi) or TIMP-3
followed by ischaemia and LPS (combi+TIMP). Following 6
hours of reperfusion, blood, bile, liver and bile duct tissue was
collected for analysis of liver function, biliary injury and expression
of genes encoding for bile acid transporters and cytokines.
Results TIMP-3 treatment prior to liver ischaemia and LPS administration
resulted in a decreased bile duct injury severity score
compared to non-pre-treated animals (sham: 0.00 (0.00-3.00);
TIMP: 0.00 (0.00–4.00); combi: 4.50 (1.00-6.00); combi+TIMP:
2.00 (0.00–5.00)). This was further supported by a
significant decrease in lactate dehydrogenase in bile, a marker
of biliary injury (combi: 15.78±9.46; combi+TIMP: 6.54±2.33
U/L/ gram wet liver weight p=0.007). Despite improvement
of biliary injury, hepatocellular injury remained unaltered as
evidenced by increased serum alanine transaminase in both
the combi and combi+TIMP groups (sham: 40.90±9.63;
combi: 109.64±40.96; combi+TIMP: 114.63±84.23, p:
0.03 and 0.024 respectively). Bile flow was normalised following
TIMP pre-treatment (combi: 64.20±14.57; combi+LPS:
85.37±10.99, p=0.012), as were gene expression levels of
CYP27a1 and CYP7b1 (2 fold and 1,5 fold reduction, p=0.01
and 0.02 respectively). This may reflect a normalisation of total
bile acid synthetic capacity. Furthermore, gene expression of
MIP-2, osteopontin and LBP were significantly down regulated
following TIMP pre-treatment (8 fold, 6 fold and 2 fold respectively,
p