studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 677A
951
Fermented soymilk prevent free fatty acid-induced lipogenesis
and production of reactive oxygen species in
hepatocellular steatosis model
Sang Bong Ahn 1 , Byoung Kwan Son 1 , Dae Won Jun 2 , Yong Kyun
Cho 3 ; 1 Gastroenterology, Eulji University, School of Medicine,
Seoul, Korea (the Republic of); 2 Internal Medicine, Hanyang
University, College of Medicine, Seoul, Korea (the Republic of);
3 Internal Medicine, Kangbuk Samsung Hospital, Sungkyunkwan
University, School of Medicine, Seoul, Korea (the Republic of)
INTRODUCTION: Ingredients of soy and fermented products
have been widely utilized as food supplement for health-enhancing
properties, such as reducing the risk of osteoporosis,
protection of cardiovascular diseases, and prevention of prostate
and breast cancer. This study was carried out to examine
the effects of fermented soymilk (FSM) on the free fatty acid-induced
lipogenesis in an in vitro model of hepatocellular steatosis
model. MATERIALS AND METHODS: HepG2 cells were
incubated with 0.2 mM of palmitic acid (PA) for 24 h to induce
lipogenesis and to accumulate the intracellular lipid accumulation,
which was observed by oil red O and Nile red staining.
The PA treated cells were co-incubated with 0.04~1.0% of
lyophilized FSM, 0.05 mM of genistein, and 50 nM of estrogen,
respectively. Western blot analysis of sterol regulatory element-binding
protein-1 (SREBP-1) and nuclear factor erythroid
2-related factor-2 (NRF-2) were performed to examine the lipogenesis
related extracellular signal-regulated kinase (ERK) pathway.
Cellular reactive oxygen species (ROS) was measured
by the DCFDA assay kit. RESULTS: Lipid accumulations in the
PA and FSM co-incubated cells were significantly decreased
by 0.5% and 1.0% of FSM without cytotoxicity. Treatments of
PA and combining with genistein and estrogen significantly
increased the expressions of SREBP-1. However, FSM co-incubation
significantly attenuated the expression of SREBP-1
in the PA treated cells. In addition, expression of NRF-2 and
phosphorylation of ERK were significantly increased in the PA
and FSM co-incubated cells. PA induced ROS production was
significantly reduced by 1.0% of FSM. Meanwhile, genistein
or estrogen alone did not lead to significant differences in ROS
production. CONCLUSION: Our results show that bioactive
components, except genistein and phytoestrogen, in fermented
soymilk protect hepatocytes against lipid accumulation and
ROS production induced by free fatty acid. These effects may
be mediated by inhibition of SREBP-1 and activation of NFR-2
via ERK pathway in hepatocytes.
Disclosures:
The following authors have nothing to disclose: Sang Bong Ahn, Byoung Kwan
Son, Dae Won Jun, Yong Kyun Cho
952
Increased 53-binding protein 1 nuclear focus expression
in patients with non-alcoholic steatohepatitis
Koji Okamoto 2 , Yuko Akazawa 4 , Hisamitsu Miyaaki 4 , Satoshi
Miuma 4 , Naota Taura 4 , Hisayoshi Kondo 3 , Masahiro Nakashima 1 ,
Kazuhiko Nakao 4 ; 1 Department of tumor and diagnostic pathology,
Atomic bomb disease institute, Nagasaki, Japan; 2 Nagasaki
University School of Medicine, Nagasaki, Japan; 3 Biostatistics Section,
Division of Scientific Data Registry, Department of Radioisotope
Medicine, Atomic Bomb Disease Institute, Nagasaki, Japan;
4 Gastroenterology and Hepatology, Nagasaki University Hospital,
Nagasaki, Japan
Background : Pathogenesis of Non-alcoholic fatty liver disease
(NAFLD) and the mechanisms of HCC development remain
largely unclear. Elevated DNA damage response can result
in genomic instability, which may lead to transformation to
cancer. The p53-binding protein 1 (53-BP1) localizes at the
sites of DNA double-strand breaks by irradiation and rapidly
forms nuclear foci. We have reported that increased number
and size of 53-BP1 nuclear foci expression represents the level
of sporadic genomic instability during cervical carcinogenesis
(Matsuda K, Histopathology. 2011,441-51). However, the
presence of genomic instability in the NAFLD liver, especially
the expression pattern of 53-BP1, is largely uninvestigated.
Our aim was to assess the double-strand breaks based on
53-BP1 expression in hepatocytes of NAFLD liver and free fatty
acid-treated hepatocytes. Methods: Isolated rat primary hepatocytes
were exposed to saturated free fatty acid palmitate (200
μM) for 16 hour. In addition, 23 human liver biopsy samples,
including 5 from normal livers, 5 from simple steatotic livers,
and 13 from livers with NASH, were examined. Cells and
biopsy samples were studied by using co-immunofluorescence
with the anti-53-BP1 and hepatocyte marker (hepatocyte). The
number of 53-BP1 nuclear foci in hepatocytes and its association
with clinical features including levels of aspartate aminotransferase
(AST), alanine aminotransferase (ALT), platelet (PLT),
serum collagen type 4, and serum hyaluronic acid were then
examined. Results: The palmitate treatment in the rat primary
hepatocytes resulted in a significant increase in the number of
53-BP1 nuclear foci (6.01% vs 24.5%, control vs palmitate,
p< 0.05). Furthermore, in the human liver biopsy tissue samples,
the number of 53-BP1 nuclear foci in the hepatocytes was
increased by approximately 6 folds in the NASH livers compared
to the simple steatotic livers, and approximately 10 folds
compared to normal controls (p < 0.01). 53-BP1 nuclear foci
were evenly distributed throughout zone 1 (portal area), zone
2, and zone 3 (hepatic vein area) in the NASH liver. The rate
of hepatocytes with more than two 53-BP1 foci/ nucleus was
positively associated with age (p