LF).Resources AND Solutions
Animal care and use statement
The animal protocol was developed to minimize discomfort
or discomfort on the animals. The animals had been
acclimatized to laboratory circumstances (23 ��, 12 h/12 h light/dark, 50% humidity, ad libitum entry to food
and water) for two weeks before experimentation.
Intragastric gavage administration was carried out
with conscious animals, Camptothecin UNC1999 Paclitaxel utilizing straight gavage needles
ideal for the animal size (15-17 g body weight:
22 gauge, 1 inch length, 1.25 mm ball diameter).
All animals had been euthanized by barbiturate overdose
(intravenous injection, 150 mg/kg pentobarbital
sodium) for tissue assortment.
Animals and chemical compounds
This analysis was implemented following the
suggestions from the Guidebook for the Care and Use
of Laboratory Animals of the Ministry of Health and fitness in the
People��s Republic of China.
The protocol was accepted
from the Committee around the Ethics of Animal Experiments
of Guangdong Health care University (Permit Variety: SYXK
2008-0007). Male C57BL/6 mice which were 8 wk old
(purchased from Shanghai Slac Laboratory Animal
Corporation and stored in SPF environment) had been utilised
in this study.
Major materials employed in this investigation incorporated
DHM and CCl4 (Sigma-aldrich, St Louis, United
States), assay kits for detection of serum ALT, AST, albumin, superoxide dismutase (SOD) and tissue SOD
(Jiancheng, Nanjing, China), assay kits for detection
of serum IL-1��, IL-6 and TNF-�� (RD corporation,
Minneapolis, United states of america), mouse monoclonal
antibody to proliferating cell nuclear antigen (PCNA)
and SABC staining kit (Boster, Wuhan, China), key
antibodies to JNK, phosphorylation-JNK, TNF-��,
Cytochrome C, Bax and ��-actin (Cell signaling, Beverly,
MA), colorimetric assay kits of Caspases-3, 6, 8, and 9
activities (Calbiochem, La Jolla, CA), In Suit Cell Death
Detection kit-POD (Roche, Basal, Switzerland), and
SP600125 (Selleckchem, Houston, U.s.).
Preparation of animal model and DHM administration
The preparation in the animal model was done as
. Briefly, acute liver injury
was induced in mice by intraperitoneal injection of CCl4
(1 mL/kg) to test the hepatoprotective function of DHM.
Meanwhile, ALF mice had been ready by intraperitoneal
injection of a lethal dose of CCl4 (2.
6 mL/kg) to test
the impact of DHM about the survival rate of mice
was dissolved in 0.5% sodium carboxymethylcellulose (CMC-Na) diluted in ultrapure water, to a ultimate concen-
tration of 37.5 mg/mL. Then, mice had been handled orally
with DHM (150 mg/kg per Camptothecin UNC1999 Paclitaxel mouse; once each day, for 4
d) 2 h soon after CCl4 treatment method. JNK inhibitor SP600125 (50
mg/kg) was administered by intraperitoneal injection
1 h before DHM treatment
. Mice were dedicated on
days 1, 2, 3, 5 and 7 after CCl4 treatment to examine
a series of indicators of liver damage and regeneration.
Serum AST, ALT, albumin, SOD, IL-1��, IL-6 and TNF-��
Serum AST, ALT, albumin and SOD levels were established with the commercial assay kits. Serum
IL-6, IL-1�� and TNF-�� levels were