Protective effects of modafinil against α-naphthylisothiocyanate-induced cholestatic liver injury in mice: insights into inflammation and apoptosis modulation

Abstract This study examined the protective effects of the central nervous system stimulant modafinil (MODA) in a mouse model of cholestatic hepatitis caused by α-naphthylisothiocyanate (ANT). Cholestatic hepatitis is a liver disease that is characterized by liver damage, discomfort, and jaundice. MODA was selected because of its known anti-inflammatory, anti-apoptotic, and protective qualities. For the study, mice received ANT (50 mg/kg) to induce liver injury, while MODA (60 mg/kg) was administered daily for five days. Liver function was assessed by measuring plasma levels of ALT, AST, and total bilirubin, alongside markers of oxidative stress. Apoptotic and inflammatory indicators, including p53, Bax, Bcl-2, caspase-3, and NF-κB, were quantified using immunohistochemistry and ELISA. Caspase-1, -3, and -9 activities were also measured. Results showed that ANT caused hepatomegaly, increased oxidative stress, inflammation, and apoptosis in the liver. However, MODA treatment significantly reduced these adverse effects. Specifically, MODA decreased liver enlargement, improved histological damage, lowered ALT and AST levels, and enhanced oxidative balance by reducing MDA and increasing GSH and SOD. Inflammatory mediators like NF-κB, IL-6, TNF-α, and caspase-1 were also suppressed. Moreover, MODA modulated apoptotic pathways, leading to decreased Bax and caspase-3/9 expression and increased Bcl-2 expression. In conclusion, MODA offers protective benefits against ANT-induced cholestatic liver injury by mitigating oxidative stress, inflammation, and mitochondria-mediated apoptosis. These findings highlight MODA's therapeutic potential for cholestatic hepatitis, suggesting a quicker path to clinical application given its existing approval for other uses. However, MODA retained only mild ability to restore normal bilirubin clearance. Due to the acute nature of the model, the anti-fibrotic potential in a chronic disease model should be evaluated in future, thereby facilitating advancement in clinical translation.