studies

266A AASLD ABSTRACTS HEPATOLOGY, October, 2015
ASH. Inhibition of peroxisome proliferator-activated receptor
g (PPARg) or cyclic-AMP-responsive-element binding protein H
(CREBH) attenuated chronic-plus-binge ethanol-induced elevation
of Fsp27a and FSP27b mRNA, respectively, and subsequently
ameliorated liver injury. Overexpression of Fsp27 and
ethanol exposure synergistically induced mitochondrial reactive
oxygen species production and hepatocyte injury in vivo and
in vitro, which is likely owing to the mitochondrial location of
FSP27 leading to the decreased mitochondrial complex I activity.
Finally, hepatic expression of CIDEC mRNA was elevated
and positively correlated with hepatic steatosis, disease severity,
and mortality in AH patients. Conclusion: FSP27/CIDEC
gene product promotes ASH in chronic-plus-binge ethanol-fed
mice and in human AH. Targeting CIDEC gene is likely to be a
novel therapeutic targets for the treatment of ASH.
overgrowth was similar to WT mice, alcohol-fed Reg3b and
Reg3g deficient mice showed a significantly higher number of
mucosa-associated bacteria in the small intestine as compared
with their respective WT littermates. This was accompanied
by a significantly increased number of bacteria translocating
through intestinal epithelial cells and more positive mesenteric
lymph node cultures in alcohol-fed Reg3b -/- and Reg3g -/- mice.
Interestingly, absence of Reg3b or Reg3g did not affect the
intestinal paracellular barrier function as determined by systemic
LPS level and fecal albumin. Most importantly, Reg3b
and Reg3g deficient mice showed more alcoholic liver disease
as assessed by higher plasma ALT levels, increased hepatic
triglycerides and inflammation. To corroborate our data, we
generated transgenic mice overexpressing Reg3g under the
control of the intestine specific villin promoter. Consistent with
our results, Reg3g transgenic mice showed significantly less
mucosa-associated bacteria than WT littermates after alcohol
feeding. Overexpression of intestinal Reg3g suppressed
translocation of bacteria through intestinal epithelial cells to
mesenteric lymph nodes, while paracellular permeability and
systemic LPS levels were not changed after alcohol feeding.
Reg3g transgenic mice were protected from alcohol-induced
liver injury, steatosis and inflammation. Conclusion: Intestinal
Reg3b and Reg3g prevent bacterial colonization of epithelial
surfaces, which reduces translocation of viable bacteria and
prevents alcoholic liver disease. Reg3 proteins are novel and
promising targets in the treatment of alcohol-induced liver disease.
Disclosures:
The following authors have nothing to disclose: Lirui Wang, Peng Chen, Lora V.
Hooper, Bernd Schnabl
Disclosures:
Ramon Bataller - Advisory Committees or Review Panels: Sandhill; Consulting:
VTI, Oncozyme Pharma
The following authors have nothing to disclose: Ming-Jiang Xu, Yan Cai, Hua
Wang, José T. Altamirano, Gemma Odena, Frank J. Gonzalez, Bin Gao
111
Antimicrobial proteins Reg3b and Reg3g protect mice
from alcoholic liver disease by preventing bacterial
translocation
Lirui Wang 1 , Peng Chen 1 , Lora V. Hooper 2 , Bernd Schnabl 1 ;
1 University of California San Diego, La Jolla, CA; 2 Department of
Immunology, The University of Texas Southwestern Medical Center,
Dallas, TX
Background: Antimicrobial proteins are secreted by intestinal
epithelial cells and Paneth cells. They represent the first
line of defense against pathogens and maintain homeostasis
with commensal bacteria. We have previously shown that the
expression of the C-type lectin regenerating islet derived-3
(Reg3) is suppressed in the small intestine after chronic alcohol
use in mice and humans, yet functional consequences of
lower Reg3 expression on the intestinal microbiota, bacterial
translocation and alcoholic liver disease are unknown. The
aim of our study was to investigate the role of Reg3b and
Reg3g in chronic alcoholic liver disease. Methods and Results:
A Lieber-DeCarli model was used to induce intestinal dysbiosis,
bacterial translocation and liver disease in mice. After alcohol
feeding for 8 weeks, wild-type (WT) littermate mice showed
bacterial overgrowth of the luminal and the mucosa-associated
microbiota in the small intestine. While luminal bacterial
112
Hepatocytes from mice on intragastric feeding model of
alcoholic steatohepatitis release extracellular vesicles
with specific microRNA cargo that modulate hepatic
stellate cell and macrophage phenotype
Akiko Eguchi 1 , Jihoon Kim 1 , Lucila Ohno-Machado 1 , Hidekazu
Tsukamoto 2 , Ariel E. Feldstein 1 ; 1 UCSD, La Jolla, CA; 2 USC, Los
Angeles, CA
Liver inflammation and fibrosis are key histological features
associated with prognosis in patients with alcoholic steatohepatitis
(ASH). Extracellular vesicles (EVs) are released during cell
stress or demise, can contain a barcode of the cell of origin
including specific microRNAs and are growingly recognized
as key cell-to-cell communicators. Here we tested the hypothesis
that during ASH development, hepatocyte damage release EVs
with a microRNA signature that can fuse with hepatic stellate
cells (HSC) and Kupffer cells (KC) to regulate their phenotype.
Methods: C57/B6 mice were placed on intragastric feeding
model of continuous ethanol infusion or control diet for 4 weeks
to reproduce a physiologically relevant model of ASH. The
extent of steatosis, inflammation, and fibrosis was assessed
by histological and molecular analyses on liver specimens.
Isolated hepatocytes (HC) or KC from ASH or control mice
were incubated in medium-EV free serum and HC- or KC-derived
EVs were isolated by ultracentrifugation from culture
medium. A complete characterization of EVs was performed
by FACS, electron microscopy, dynamic light scattering. HC-EV
biological function was investigated in isolated primary HSCs
or KCs derived from C57/B6 mice by cell morphology or
qPCR. Comprehensive encapsulated miRNA in HC-EVs were
assessed via transcriptome using illumina miRNA-seq. Results:
We observed highly significant differences in the levels of HCor
KC-EVs between control group and ASH (1.3x10 6 HC-EV/