studies

952A AASLD ABSTRACTS HEPATOLOGY, October, 2015
liver sections. Ascitic fluid was evaluated for bacterial growth
in thioglycolate medium. Ex-vivo liver perfusion experiments
were performed for assessing endothelium-dependent and –
independent reactivity. Results: Compared with controls, rats
with oral CCl4 gavage tended to show decreased survival and
body weight gain, both of which were further worsened by
enoxaparin 180 U/kg bw (p< 0.01). Rats with CCL4-induced
cirrhosis showed altered laboratory parameters (increased
INR, AST, ALT, bilirubin / decreased albumin, total proteins,
glucose and platelets) regardless of enoxaparin treatment. In
all experimental models, cirrhotic rats receiving saline and
those receiving enoxaparin showed similar increases in the
area of liver fibrosis compared with controls (p< 0.001). Rats
with cirrhosis induced by oral CCl4 gavage and by BDL surgery
developed increases of portal pressure and spleen-to-bw
ratios compared with control rats (p< 0.001), regardless of
enoxaparin treatment. Among rats with ascites, a similar proportion
presented positive bacterial cultures (CCl4+saline 2
of 7 vs. CCl4+enoxaparin 4 of 8, NS; BDL+saline 1 of 6 vs.
BDL+enoxaparin 2 of 7, NS). Potential effects of enoxaparin
on hepatic vascular reactivity were only observed in rats
receiving enoxaparin 180 U/kg bw from the beginning of
CCl4 administration, and consisted of increased portal venous
resistance after addition of acetylcholine or S-nitroso acetylpenicillamine
(SNAP, both p< 0.05) and increased sinusoidal
resistance after addition of SNAP (p< 0.05). Conclusion: Our
experimental data do not support a role of long-term treatment
with enoxaparin for improving liver fibrosis, portal hypertension
or endothelial dysfunction in cirrhosis.
Disclosures:
The following authors have nothing to disclose: Jose Ignacio Fortea, Alexander
Zipprich, Carolina Fernandez Mena, Christopher F. Rose, Juan Bañares, Marta
Puerto, Cristina R. Bosoi, Jorge Almagro, Marcus Hollenbach, Marc-André Clément,
Javier Vaquero, Rafael Bañares, Cristina Ripoll
1521
Neuro-inflammation in Cirrhotic Mice is Dependent on
Gut Microbial Colonization: Implications for Hepatic
Encephalopathy
Dae Joong Kang 1 , Siddhartha Ghosh 1 , Daniel Carl 1 , Arun J.
Sanyal 1 , Ryan B. Sartor 2 , Phillip B. Hylemon 1 , Huiping Zhou 1 , Runping
Liu 1 , Xiang Wang 1 , Jing Yang 1 , Chunhua Jiao 1 , Jasmohan S.
Bajaj 1 ; 1 VCU and McGuire VAMC, Richmond, VA; 2 Gnotobiotic
Rodent Research Center, University of North Carolina, Chapel
Hill, NC
Neuro-inflammation has been proposed as a mechanism for
hepatic encephalopathy (HE) but the source for this is not clear.
Specifically, brain inflammation in cirrhotic models in the germfree
mice (GF) condition has not been studied. Aim: Assess the
impact of intestinal microbiota on brain inflammation in cirrhotic
mice in a conventional (conv) and GF setting. Methods:
We used C57BL/6 mice in a conv and GF setting. Sixteen mice
were in the GF condition; of which 6 remained GF while 10
were administered CCL4 1ml/kg in sesame oil using gavage
twice/week till week 16. An age-balanced group of 10 conv
mice were included; 5 remained healthy while cirrhosis using
CCL4 was induced in 5 mice using 1ml/kg CCL4 administration
twice a week for 12 weeks. On sacrifice, brain tissue was
separated into cortex (Cx) and cerebellum (Cbl). Analysis was
performed for mRNA of inflammation (IL-6, IL-1β) and activation
of microglia and glia (IBA-1, GFAP) using qPCR (adjusted for
GADPH) in both brain regions of all mice using Kruskall-Wallis
tests. Results: All mice survived till end of the experiment. Both
CCL4-treated groups (conv & GF) showed cirrhosis on sacrifice
on visual and histological examination. Comparison within GF
group: There was no significant difference in inflammation or
microglial/glial activation in both brain regions in GF controls
vs. CCL4 GF cirrhotics (table) Comparison within Conv
group: There was a significant increase in mRNA of IL-1β,
GFAP and IBA-1 in both Cbl and Cx of CCL4 conv cirrhotic
mice compared to their conv non-cirrhotic counterparts. IL-6
was unchanged. Comparison between GF Cirr and Conv cirr:
Conv CCL4 cirrhotic mice showed a significant increase in Cbl
and Cx IL-1β, GFAP and IBA-1compared to GF cirrhotic mice.
IL-6 was again not different between groups. Conclusions: Glial
and microglial activation and induction of IL-1β occurs in cirrhotic
mice only in the presence of gut microbiota. This suggests
an essential role of the gut microbiota in the development
of neuro-inflammation in cirrhosis and could have implications
for the gut-brain axis management in hepatic encephalopathy.
Neuroinflammation and Glial/Microglial Expression between
Groups
Disclosures:
Arun J. Sanyal - Advisory Committees or Review Panels: Bristol Myers, Gilead,
Genfit, Abbott, Ikaria, Exhalenz; Consulting: Salix, Immuron, Exhalenz, Nimbus,
Genentech, Echosens, Takeda, Merck, Enanta, Zafgen, JD Pharma, Islet
Sciences; Grant/Research Support: Salix, Genentech, Intercept, Ikaria, Takeda,
GalMed, Novartis, Gilead, Tobira; Independent Contractor: UpToDate, Elsevier
Jasmohan S. Bajaj - Advisory Committees or Review Panels: Salix, Merz, otsuka,
ocera, grifols, american college of gastroenterology; Grant/Research Support:
salix, otsuka, grifols
The following authors have nothing to disclose: Dae Joong Kang, Siddhartha
Ghosh, Daniel Carl, Ryan B. Sartor, Phillip B. Hylemon, Huiping Zhou, Runping
Liu, Xiang Wang, Jing Yang, Chunhua Jiao
1522
Emricasan, a pan caspase inhibitor, improves survival
and portal hypertension in a murine model of long-term
common bile-duct ligation
Akiko Eguchi 1 , Alexander Wree 1 , Yukinori Koyama 1 , Casey Johnson
1 , Ryota Nakamura 1 , Patricia C. Contreras 2 , Ariel E. Feldstein 1 ;
1 UCSD, La Jolla, CA; 2 Conatus Pharmaceuticals, San Diego, CA
Development of portal hypertension (PTH) is a central prognostic
factor in patients with cirrhosis. Current pharmacotherapy
remains limited and novel therapies are greatly needed. Circulating
microparticles (MPs) are released by hepatocytes in
a caspase dependent manner, are increased in circulation of
patients with cirrhosis and contribute to PTH via induction of
impair vasoconstrictor responses Here we tested the hypothesis
that Emircasan (IDN-6556) a pan-caspase inhibitor ameliorates
PTH via its anti-fibrotic effects and reduction in release
of MPs. Methods: Secondary biliary cirrhosis was induced by
long-term common bile-duct ligation (CBDL) in C57BL/6 mice.
Controls were sham-operated, the abdominal cavity opened,
but no ligature placed. Mice were treated with 10 mg/kg/
day of IDN-6556 or placebo for 3 wks via i.p. injection (4
groups, n= 7 -12 in each group).. After 3 wks, portal pressure
was measured in each mouse and serum and livers were collected.
Circulating MPs were isolated and characterized using
various approaches including FACS analysis. Portal pressure
was measured using catheter from ileocolic vein. Hepatocellular
damage was assessed by liver histopathology, serum ALT
levels, and TUNEL assay. Liver fibrosis was assessed by digital
image analysis of Sirius red stained sections. Results: In