Hend Ashour; Hind Zafrah; Muataz Elsiddig Dafaalla Mohammed; Laila Ahmed Rashed; Abbas Mohamed Abbas; Samaa Samir Kamar & Asmaa Mohammed ShamsEldeen

Cholestasis is a leading cause of hepatic fibrosis. The herbal plant thymoquinone (TQ) has a well-known hepatoprotective effect against liver fibrosis. In this study, we investigated the role of ferroptosis in TQ-mediated antifibrotic action in a rat model of extrahepatic cholestasis developed secondary to bile duct ligation (BDL). Four groups (n=10) of adult male Wistar albino rats were included in the study; 1) control, 2) BDL model, 3) treated group with 50 mg.kg-1.day-1 TQ, and group 4) rats in this group were treated with TQ and the ferroptosis inhibitor, ferrostatin-1 (Fer-1) at a dose of 1 mg.kg-1.day-1. TQ treatment effectively attenuated the inflammation and fibrosis detected in the BDL group by modulating (P<0.01), NF-κB, TNF-α, interleukins; IL-6, 10, TGFβ-1, collagen type I, III. TQ improved liver function which was confirmed by histological H&E and Sirius red stain. TQ augmented (p<0.001) the heme oxygenase (HO-1) protein levels about 4 fold compared to the BDL group suggesting its role in hepatic protection. TQ stimulated the ferroptosis process and increased glutathione peroxidase (GPX4), Iron content, and the transferrin receptor (TfR-1). Fer- 1 addition to TQ did not affect the anti-inflammatory actions of TQ. However, it significantly (p<0.001) attenuated its antifibrotic property. In conclusion, The antifibrotic effect of Thymoquinone was partially inhibited secondary to Fer-1. Thus, documenting the mediating role of ferroptosis in the TQ protection against hepatic fibrosis.

KEY WORDS: Liver fibrosis; Cholestasis; Ferroptosis; Ferrostatin; Thymoquinone.a

ASHOUR, H.; ZAFRAH, H.; MOHAMMED, M. E. D.; RASHED, L. A.; ABBAS, A. M.; KAMAR, S. S. & SHAMSELDEEN, A. M. Ferrostatin-1 partially suppressed the anti-fibrotic actions of thymoquinone in a rat model of cholestasis-induced liver injury. Int. J. Morphol., 43(3):1001-1010, 2025.