Ab263882

Fifteen C57BL/6 J male mice (6–8 weeks) were provided by Charles River Lab (Beijing, China). Before the start of the experiment, the animals were placed in an air-conditioned room at 25°C with a 12-hour light cycle. All animals were given proper care during the study. The experiment was approved by the Suzhou Ninth People’s Hospital Ethics Committee (Approval No. KY2021-049-01) and followed the ethical guidelines on the care and use of laboratory animals for this research.
Liver fibrosis model in mice was constructed through twice-weekly intraperitoneal injections of CCl₄ (25%, 5 ml/kg) for 8 weeks. Mice were randomly assigned into three groups: Control group, CCl₄ group and CCl₄+ TMZ group (40 mg/kg, by gavage). After euthanasia of the mice, the livers were stripped, and half in liquid nitrogen were frozen rapidly and kept at −80°C. The other half were fixed in 4% paraformaldehyde sections for 24 h and paraffin embedded. Levels of liver enzyme including alanine aminotransferase (ALT; ab282882) and aspartate aminotransferase (AST; ab263882) in plasma samples were determined using corresponding ELISA kits from Abcam (Shanghai, China) in the light of the manufacturer’s guidance. For a more visual analysis of liver fibrosis, tissue sections were analyzed using Masson’s Trichrome Stain kit (Beijing Solarbio Technology Co., Ltd. China) according to the experiment manual.

The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were examined at 6h after GalN/LPS stimulation according to the ALT and AST ELISA kit instructions (ab282882 and ab263882, Abcam, Cambridge, UK). The concentrations of MDA and GSH-Px in the mouse plasma from all the groups were determined by ELISA analysis. The experiment was conducted using a commercially available kit following the manufacturer’s instructions (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).

The ELISA kits of mouse intracellular adhesion molecule-1 (ICAM-1, ab252355, Abcam), vascular cell adhesion molecule-1 (VCAM-1, ab201278, Abcam), alanine aminotransferase (ALT, ab282882, Abcam), aspartate aminotransferase (AST, ab263882, Abcam), and creatinine (CREA, KT21312, Mskbio) were used to detect the expression levels of ICAM-1, VCAM-1, ALT, AST, and CREA, respectively, in the serum according to the instructions of the manufacturer. Each serum sample was tested twice to limit the chance of error, and the average value was recorded.