The Protective Effects of Crocin and Pentoxifylline, Alone and in Combination, in Alleviating Acrylamide‐Induced Hepatic Injury in NMRI Mice

Acrylamide (ACR) is formed as a byproduct during high-temperature processing of carbohydrate-rich, low-protein foods and represents a widespread dietary contaminant. Given the extensive daily human exposure, its hepatotoxic potential is a growing concern. Therefore, this study aimed to evaluate the protective effects of crocin and pentoxifylline (PTX), administered individually or in combination, against ACR-induced hepatotoxicity in an animal model. Thirty male NMRI mice were randomly assigned to five groups (n = 6): normal, ACR, ACR + crocin, ACR + PTX, and ACR + crocin + PTX. Crocin (50 mg/kg) and PTX (50 mg/kg) were administered orally for 2 weeks. Hepatotoxicity was induced by a single intraperitoneal injection of ACR (10 μg/kg), and animals were euthanized 24 h later. Serum samples were collected for the assessment of liver function, oxidative stress, antioxidant defense, and inflammatory markers, while liver tissues were subjected to histopathological and molecular analyses. Crocin significantly reduced IL-6 (~19%) and increased CAT activity (~14%), whereas PTX decreased TNF-α levels and elevated GSH (~31.7%). In contrast, the combined crocin + PTX treatment exerted more pronounced protective effects, significantly reducing ALT and AST levels, lowering TNF-α (~31.25%) and IL-6 (~20.7%), and attenuating oxidative stress through reductions in MDA (~19.75%) and TOS (~17.5%), accompanied by a marked increase in TAC (~63.2%). This combination also enhanced CAT activity (~25%) and GSH levels (~44.4%), although SOD activity remained unchanged. At the gene expression level, none of the interventions fully restored Nrf2 or FOXO3a expression; however, PTX significantly upregulated FOXO3a expression, resulting in a ~26-fold increase compared to the ACR group, although levels remained markedly lower than those of the normal group. Histopathological evaluation revealed reduced hepatic damage in the treatment groups. Overall, these findings suggest that while crocin and PTX individually confer limited protection, their combined administration exerts a significant synergistic hepatoprotective effect against ACR-induced toxicity.