New insight on the acute CCl₄-induced hepatotoxicity model in rats

The CCl₄-induced hepatotoxicity model is a traditional preclinical assay applied to evaluate potential hepatoprotective compounds. However, several studies have used it with inappropriate dose and exposure time, generating both weak response or irreversible liver injury, as well as lack of representative liver and plasma biomarkers. Therefore, this study aims to determine the best dose and exposure time of CCl₄ in Wistar rats, permitting a proper evaluation of potential hepatoprotective effect. Thus, CCl₄-intraperitoneal doses of 0.5, 1.0, and 2.0 mL/kg were first evaluated 24 h post-exposure, and then with the best dose achieved, it was also assessed at 6 and 12 h post-exposure. The determination of the main hepatotoxicity biomarkers, including malondialdehyde (MDA), aspartate transaminase (AST), and alanine transaminase (ALT), and histopathological analyses were performed. The results suggest that 6h CCl₄ post-exposure is too short to induce ideal liver injury, and at 24 h, a suggestive rat free-radical scavenger mechanism seems to revert CCl₄-initiated damage. According to these data, the ideal acute CCl₄-induced hepatotoxicity model was established at a dose of 2.0 mL/kg and 12 h post-exposure in Wistar rats, which demonstrated a significant increase of liver MDA levels without irreversible injury, permitting a proper and reliable evaluation of potential hepatoprotective compounds.