Liver tissue (60 mg) was mixed with
1.0mL
of 6 N NaOH and incubated at 100°C for 20
min
with frequent vigorous shaking. After cooling, 1.0 mL of
3N
HCl was added to the solution, the pH was regulated to
6.0~6.8, and then 10 mL of water was added and mixed.
Then, 3.0 mL of the suspension was added to 20~30 mg
active carbon, and the sample was mixed and centrifuged at 3500 rpm for
10 min. The supernatant was taken to measure the hydroxyproline contents
using a colorimetric hydroxyproline test kit according to the
manufacturer’s instructions (Institute of Jiancheng Bioengineering,
Nanjing, China). The hydroxyproline content was expressed as micrograms
per gram of liver weight (μg/g liver).
Hepatic histological and
pathological