studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 685A
was not observed. PA also induce PINK1 cleavage, implying
the possible inflammatory and apoptosis signaling. Using specific
protease inhibitor that cleaves PINK1 in inner mitochondrial
membrane, we were able to block its cleavage which
also leads to the inhibition of apoptosis. As mitophagy does not
occur which is important mitochondrial quality control, this lead
to surplus damaged mitochondria and subsequent apoptosis
of cell further triggering inflammation. Conclusion: Fatty acids
induce aberrant mitochondrial dynamics involving mitochondrial
fission, caused apoptosis but does not lead to mitophagy.
Disclosures:
The following authors have nothing to disclose: Muhammad Sohail, Seong-Jun
Kim, Aleem Siddiqui
Figure: NFκB inhibition by AP inhibits IL-1β expression induced by
LPS. (A) AP treatment inhibited NFκB activation by LPS in HepG2
cells, evaluated through a reporter gene assay. (B) NFκB inhibition
by AP treatment inhibited IL1β induction by LPS in Palmitic
acid-treated HepG2 cells. (**p≤0.01, ***p ≤ 0.001).
969
NFkB inhibition by Andrographolide ameliorates
inflammation and fibrogenesis through inflammasome
substrate depletion in experimental Non-Alcoholic Steatohepatitis
(NASH)
Daniel Cabrera 1,2 , Alexander Wree 3 , Davide Povero 3 , Nancy
Solis 1 , Margarita Pizarro 1 , Pamela Rojas de Santiago 1 , Javiera
Torres 7 , Han Moshage 5 , Claudio Cabello-Verrugio 4 , Ariel E.
Feldstein 3 , Enrique Brandan 6 , Marco Arrese 1 ; 1 Departamento de
Gastroenterología, Facultad de Medicina Pontificia Universidad
Católica de Chile, Santiago, Chile; 2 Departamento de Ciencias
Químico-Biológicas, Universidad Bernardo O Higgins, Santiago,
Chile; 3 Department of Pediatrics, University of California San
Diego, San Diego, CA; 4 Departamento de Ciencias Biológicas,
Universidad Andres Bello, Santiago, Chile; 5 Department of Gastroenterology
and Hepatology, University of Groningen, Santiago,
Chile; 6 Departamenti de Biologia Celular y Molecular, Pontificia
Universidad Católica de Chile, Santiago, Chile; 7 Departamento
de Anatomia Patologica, Pontificia Universidad Católica de Chile,
Santiago, Chile
Background: Options to treat NASH are limited. Andrographolide
(AP), the bioactive component of Andrographis paniculata,
has potent anti-inflammatory activity related to NFκB
inhibition. NFkB is a crucial factor in Interleukin-1β (IL-1β)
expression, the main substrate for the inflammasome. Aim: To
evaluate the effects of AP in experimental NASH and its influence
in inflammasome activity. Methods: C57bl6 mice were
fed a choline-deficient-amino-acid–defined (CDAA) diet (22
weeks) with or without AP (1 mg/Kg, 3 times/week, intraperitoneally).
Serum levels of alanine aminotransferase (ALT) and
hepatic steatosis, inflammation, and fibrosis were assessed.
Hepatic triglyceride content (HTC) and hepatic mRNA levels
of selected pro-inflammatory and pro-fibrotic genes as well as
those related to inflammasome were also measured. Direct AP
effect on IL-1β expression and inflammasome activity was evaluated
in HepG2 cells loaded with a mixture of FFAs during 24h.
Results: AP decreased HTC and attenuated hepatic inflammation
and fibrosis in CDAA-fed mice reducing serum ALT activity
and hepatic collagen deposition. AP treatment was associated
with a strong reduction in hepatic macrophage infiltration
and of hepatic mRNA levels of both pro-inflammatory and
pro-fibrotic genes. In addition, mice treated with AP showed
reduced expression of inflammasome genes (-60% reduction in
inflammasome adaptor ASC hepatic mRNA levels, and -40%
reduction in NLRP3 hepatic mRNA levels). Finally, AP inhibited
IL-1β expression induced by LPS through NFκB inhibition in fat
laden HepG2 cells (figure). Conclusion: AP administration prevented
liver damage in NASH. Inflammasome inactivation by
NFκB-dependent mechanism involving IL-1β expression inhibition
is likely involved in the therapeutic effects of AP. (Fondecyt
1150327 to M.A. and FONDECYT PD3140396 to D.C.)
Disclosures:
The following authors have nothing to disclose: Daniel Cabrera, Alexander
Wree, Davide Povero, Nancy Solis, Margarita Pizarro, Pamela Rojas de Santiago,
Javiera Torres, Han Moshage, Claudio Cabello-Verrugio, Ariel E. Feldstein,
Enrique Brandan, Marco Arrese
970
Reduction of Hepatic 27-Hydroxycholesterol in Steatohepatitis
Model Mice with Insulin Resistance
Sho-ichiro Yara 1 , Tadashi Ikegami 1 , Akira Honda 1,2 , Teruo
Miyazaki 2 , Masashi Murakami 1 , Junichi Iwamoto 1 , Yasushi Matsuzaki
1 ; 1 Division of Gastroenterology and Hepatology, Tokyo
Medical University Ibaraki Medical Center, Ami, Inashiki, Japan;
2 Collaborative Research Center, Tokyo Medical University Ibaraki
Medical Center, Ami, Inashiki, Japan
BACKGROUND: 27-hydroxycholesterol (HC), one of the major
oxysterol found in the human circulation, is produced by the
mitochondrial enzyme CYP27A1. Previous report demonstrated
that hepatic inflammation was increased by genetic deletion of
CYP27A1 and decreased by 27HC treatment in high fat diet
(HFD) fed steatotic mice, however, direct cause of dysregulation
of CYP27A1 and 27HC in steatohepatitis is unknown.
AIM: The current study was undertaken to examine the change
of hepatic OS levels in the Stelic Animal Model (STAM) mice,
a validated and widely used as NASH-derived HCC model.
METHODS: Extracted sterols in the liver tissue homogenates
were determined by LC-MS/MS. RESULTS: In STAM mice, IR
was induced by injection of streptozotocin at the birth period.
Four weeks after injection, feeding of high-fat diet was started,
and given over time. At week 20, STAM mice developed hepatocellular
carcinoma (HCC) whereas HCCs were not found in
control (CTL) as well as mice only fed with HFD. (1) Concentration
of hepatic cholesterol (CHOL): Hepatic CHOL concentrations
were kept in the narrow range (2.5 to 2.95 mg/mg wet
weight liver (ww)) by 12 weeks old either in CTL, HFD fed, or
STAM mice, but significantly elevated at 20 weeks in HFD fed
mice (4.7 mg/mg ww) and STAM mice (4.2 mg/mg ww) compared
to CTL (2.2 mg/mg ww). (2) Concentration of hepatic
bile acids (BAs): In HFD fed mice, hepatic BA concentrations
(111.1 to 120.8 nmol/g ww) were significantly lower than CTL
(177.4 to 205.6 nmol/g ww) over time, suggesting enhanced
BA excretion from liver under the condition with higher hepatic
CHOL level in the absence with IR. In STAM mice, hepatic BA
concentrations were maintained in a similar level of HFD at
earlier phase (4 to 8 wk old), but those were significantly elevated
at later phase (12 to 20wk old)(215.0 nmol/g ww at 12
wk, 289.0 nmol/g ww at 20wk). (3) Concentration of hepatic
OS: Among major OS tested (24S, 25epo-HC, 22RHC, 24HC,
25HC, 27HC, 4βHC), only 27HC was significantly lower in
STAM mice (0.388 to 0.579 ng/mg ww) compared to either