studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 675A
947
Adipose tissue-derived stromal cells preventatively suppressed
pathological progression of murine non-alcoholic
steatohepatitis model.
Masatoshi Yamato 1,2 , Yoshio Sakai 1 , Hatsune Mochida 2 , Akihiro
Seki 2 , Kosuke Ishida 2 , Masao Honda 2,1 , Shuichi Kaneko 2,1 ; 1 Gastoroenterology,
Kanazawa University, Kanazawa, Japan; 2 Disease
Control and Homeostasis, Kanazawa University, Kanazawa,
Japan
Non-alcoholic steatohepatitis (NASH) is characterized by steatosis
in hepatocytes accompanied with persisting inflammation,
developing fibrosis to ultimate cirrhosis. The pathogenesis
of NASH is not fully elucidated, and the fundamental treatment
of NASH is yet to be established. We previously reported that
ADSCs are therapeutically beneficial as being functionally and
histologically repairable of the end-stage cirrhotic mice caused
by NASH. In this study, we analyzed pathological progression
of NASH mice model and succeedingly assessed how
ADSC administration affected on early stage NASH disease
progression.[Materials and Methods] C57Bl/6J mice (female,
10 weeks old) were initiated to be fed with high fat atherogenic
(HF-AT) diet to develop steatohepatitis. On week 1, 2,
3, 4, 8, and 12, we obtained liver tissue samples. ADSCs
were isolated from adipose tissue of C57BL/6J mice by collagenase
I digestion, cultured and expanded. On week 4 and 8,
ADSCs (1×10^5 ) were injected twice every 4 weeks into the
splenic subcapsule, and on week 12, the liver tissue samples
were obtained. These liver tissues were analyzed for gene
expressions by real-time detection PCR (RTD-PCR) and immunohistochemistry.[Results]Hepatocyte
steatosis was observed
from 1 week after initiation of HF-AT feeding, not accompanied
with infiltration of inflammatory cells until 4 week. On week 4,
Gr-1+ inflammatory cells were initially found to be infiltrated
into the portal area and had continuously increased with time
until week 12. TNF-α gene expression of the liver tissue was
also increased consistently with increase of hepatic infiltration
of Gr-1+ cells. Obvious fibrosis had been observed since
week 8. After 8 week, liver fibrosis was extended, and gene
expressions of albumin and type IVa collagen was decreased
and increased, respectively. When ADSCs were injected into
NASH mice, the fibrosis area was less extended on week 12
than that of PBS injection mice group. RTD-PCR analysis of
hepatic tissue showed the substantial maintained albumin gene
expression (p=0.04) and the significant decrease level of Collagena
(p=0.04) and TNFα(p=0.04) gene expressions, which
suggested that ADSC improved protein production, suppressing
fibrosis progression, and hepatic inflammation.[Conclusion]
ADSC treatment prevented pathological progression of liver
fibrosis with persisting hepatic inflammation, and prevented the
decrease of albumin production of the liver during early phase
of NASH development. ADSC treatment is considered to be
preventively and therapeutically efficacious in part to NASH.
Disclosures:
Shuichi Kaneko - Grant/Research Support: MDS, Co., Inc, Chugai Pharma., Co.,
Inc, Toray Co., Inc, Daiichi Sankyo., Co., Inc, Dainippon Sumitomo, Co., Inc,
Ajinomoto Co., Inc, Bristol Myers Squibb., Inc, Pfizer., Co., Inc, Astellas., Inc,
Takeda., Co., Inc, Otsuka„ÄÄPharmaceutical, Co., Inc, Eizai Co., Inc, Bayer
Japan, Eli lilly Japan
The following authors have nothing to disclose: Masatoshi Yamato, Yoshio Sakai,
Hatsune Mochida, Akihiro Seki, Kosuke Ishida, Masao Honda
948
High and Low Capacity Runner (HCR and LCR) rat lines
do not display significant differences in NASH phenotype
when fed a fast food diet
Howard C. Masuoka 1,3 , Oscar Cummings 2 , Naga P. Chalasani 1,3 ,
Lawrence Lumeng 1 ; 1 Medicine, Division of Digestive and Liver DIsorders,
Indiana University, Indianapolis, IN; 2 Pathology and Laboratory
Medicine, Indiana University, Indianapolis, IN; 3 Cellular
and Integrative Physiology, Indiana University, Indianapolis, IN
Exercise and dietary changes are cornerstones of treatment
of non-alcoholic fatty liver disease (NAFLD) including non-alcoholic
steatohepatitis (NASH). Sedentary lifestyle is felt to
predispose individuals to NAFLD/NASH, and improved cardiorespiratory
fitness is associated with lower hepatic steatosis.
High and Low Capacity Runner (HCR and LCR) are established,
extensively phenotyped rat lines generated by selective breeding
for contrasting extremes in maximum aerobic-exercise
capacity (MAC). Relative to the HCR, LCR rats exhibit greater
visceral adiposity, insulin resistance, and levels of circulating
pro-inflammatory hormones but lower home cage activity, and
non-exercise activity thermogenesis. We hypothesized that LCR
rats will display increased features of NASH relatively to HCR
rats when fed a “fast food” (FF) diet high in saturated fat,
cholesterol and fructose. In mice this diet better recapitulates
features of human NASH compared to a high fat diet. We had
custom formulated a FF diet containing a caloric density of 4.61
kcal/gram with a caloric composition of 44.9% fat, 15.1%
protein, and 39.9% carbohydrates. The diet was nutritionally
complete, and contained 30% fructose, 14% lard, 8% coconut
oil, and 2% cholesterol. For 16 weeks 10 LCR and 10 HCR
male rats were fed the FF diet ad libitum. Fasting weights were
obtained weekly. Fasting serum ALT, glucose and insulin were
measured every 4 weeks. MRI images were obtained monthly
to determine adipose, lean, and fluid mass. As expected, LCR
rats gained significantly more weight than HCR rats (P