2.7. CCl4‐induced liver fibrosis

The study was performed by Pharmalegacy with approval from local ethics committee. Sprague Dawley rats were orally administered with 0.25 μL/g Carbon tetrachloride (CCl₄) in olive oil solution, starting from day 0, 3 times per week for 6 weeks. Animals were killed 48 hours after the last CCl₄ administration. PXS‐5153A was given by oral gavage after 3 weeks of CCl₄ administration and continued throughout the remainder of the study at 3 mg/kg (low dose) or 10 mg/kg (high dose) once a day or 10 mg/kg (high dose) three times a week. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were assessed in the plasma. One lobe of the liver tissue was fixed in 10% formalin, stained for Sirius red and the percentage coverage area was measured. The remainder of the liver was snap frozen and used for protein, hydroxyproline and crosslink analysis.