Khella (Ammi visnaga (L.) Lam.) and celery (Apium graveolens L.) are two valuable medicinal plant species with a wide-ranging health and therapeutic benefits, including hepatoprotective and antiviral properties. Thus, in this work, the total ethanolic extracts of the roots of the aforementioned species were studied for their antiviral potential against Hepatitis C virus (HCV) using the in vitro luciferase assay. The obtained data demonstrated the noteworthy inhibitory activity of khella roots against HCV (IC50= 9.5 µg/ml), while no noticeable inhibition was shown by celery roots. Besides, three phytosterols (I‒III) and one furanochromone (IV), with previously reported antiviral properties, were isolated and identified from khella roots for the first time. These data could pave the way towards further phytochemical and biological investigation of such medicinal roots as potential sources of natural anti-HCV agents.