Abstract
Abstract
Introduction: Cholestasis is a pathological condition characterized by impaired bile flow and hepatic accumulation of toxic bile acids, leading to oxidative stress, inflammation, and cellular death. Alpha-naphthylisothiocyanate (ANIT) is widely used to induce intrahepatic cholestasis in rodent models. Emerging evidence highlights the involvement of ferritinophagy-mediated ferroptosis—an iron-dependent cell death driven by lipid peroxidation—alongside apoptosis in the pathogenesis of cholestatic liver injury.
Aim: This study aims to evaluate the potential protective effects and molecular mechanisms of Fisetin against ANIT-induced cholestasis in mice, focusing on the regulation of bile acid transporters, oxidative stress, inflammation, apoptosis, and ferroptosis pathways.
Methods: Fifty-six male mice were randomized into six groups (n=8): Normal control, ANIT model control (75 mg/kg), Fisetin low-dose (20 mg/kg + ANIT), Fisetin high-dose (40 mg/kg + ANIT), Fisetin per se (40 mg/kg), Liproxstatin-1 (10 mg/kg + ANIT). Treatments were administered orally for 7 consecutive days, with a single dose of ANIT given 48 hours prior to euthanasia. Serum liver enzymes (ALT, AST, ALP) and bilirubin (TBIL, DBIL, TBA) were measured. Hepatic oxidative stress (4-HNE, GSH, NRF2), inflammatory markers (TNF-α, IL-1β, NF-κB), transporters (BSEP, NTCP, FXR), ferroptosis signaling (FTH1, NCOA4, GPX4, ACSL4), and apoptotic markers (Bax/Bcl-2, cleaved caspase-3) were evaluated alongside histopathology.
Results: Fisetin will significantly attenuate ANIT-induced liver injury, as suspected by reduced serum biochemical markers and improved hepatic histology. Fisetin treatment will decrease oxidative stress and pro-inflammatory cytokines while restoring antioxidant defenses via NRF2 activation. Furthermore, Fisetin will upregulate key bile acid transporters (BSEP, NTCP, FXR) and effectively inhibited ferroptosis and ferritinophagy by downregulating NCOA4 and ACSL4 while increasing GPX4 and FTH1 expressions. Apoptotic markers will also significantly suppressed.
Conclusion: Fisetin will exert a hepatoprotective effects against ANIT-induced cholestasis by modulating bile acid transporters and suppressing inflammation, apoptosis, and ferritinophagy-mediated ferroptosis.
| Has the manuscript been published | not published |
|---|
