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