Abstract
Our study focuses mainly on identifying the hepatoprotective activity of Naregamia alata ethyl acetate (NAEA) extract on Wistar rats. In addition, tangeretin was isolated, characterized and studied for in vivo and in silico approaches. D-GalN-induced hepatotoxicity rats were treated with 200 and 400 mg/kg of NAEA and its antioxidant and hepatoprotective activity was determined. The active constituent in the extract was isolated and spectral characterization was carried out. Cytoprotective, antioxidant and hepatoprotective activity of tangeretin was analyzed using MTT assay, DCFA-ROS assay and D-GalN induced toxicity in HepG2 cells. The anti-inflammatory activity of tangeretin in D-GalN treated cells was assessed by measuring the level of IL-6 and TNF-α via qRT-PCR. Molecular docking of tangeretin with the TACE enzyme was performed using AutoDock tools. Acute toxicity study in rats shows that NAEA exhibits no toxicity up to 4000 mg/kg. Hepatoprotective activity of the extract was confirmed by histopathological analysis and liver enzymes in d-galactosamine-induced hepatotoxicity rats treated with 200 and 400 mg/kg of NAEA. Spectral characterization reveals that the active constituent is tangeretin and MTT assay reveals IC(50) of 44.14 µM. 21.25 and 42.5 µM of tangeretin show antioxidant activity in DCFH-DA staining. The level of IL-6 and TNF-α were downregulated by tangeretin in HepG2 cells pretreated with D-Galactosamine. Molecular docking studies show that tangeretin binds to TACE with the binding energy of -9.13 kcal/mol and exhibits a low inhibition constant of 204.12 nM. Our findings show that the antioxidant, anti-inflammatory and hepatoprotective activity of Naregemia alata is due to the presence of tangeretin. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10616-026-00902-2.