title: Protective role of amantadine in mitochondrial dysfunction and oxidative stress mediated by hepatitis C virus protein expression.
abstract: Amantadine is an antiviral and antiparkinsonian drug that has been evaluated in combination therapies against hepatitis C virus (HCV) infection. Controversial results have been reported concerning its efficacy, and its mechanism of action remains unclear. Data obtained in vitro suggested a role of amantadine in inhibiting HCV p7-mediated cation conductance. In keeping with the fact that mitochondria are responsible to ionic fluxes and that HCV infection impairs mitochondrial function, we investigated a potential role of amantadine in modulating mitochondrial function. Using a well-characterized inducible cell line expressing the full-length HCV polyprotein, we found that amantadine not only prevented but also rescued HCV protein-mediated mitochondrial dysfunction. Specifically, amantadine corrected (i) overload of mitochondrial Ca²⁺; (ii) inhibition of respiratory chain activity and oxidative phosphorylation; (iii) reduction of membrane potential; and (iv) overproduction of reactive oxygen species. The effects of amantadine were observed within 15 min following drug administration and confirmed in Huh-7.5 cells transfected with an infectious HCV genome. These effects were also observed in cells expressing subgenomic HCV constructs, indicating that they are not mediated or only in part mediated by p7. Single organelle analyzes carried out on isolated mouse liver mitochondria demonstrated that amantadine induces hyperpolarization of the membrane potential. Moreover, amantadine treatment increased the calcium threshold required to trigger mitochondrial permeability transition opening. In conclusion, these results support a role of amantadine in preserving cellular bioenergetics and redox homeostasis in HCV-infected cells and unveil an effect of the drug which might be exploited for a broader therapeutic utilization.
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, via L. Pinto c/o OO.RR., 71100 Foggia, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Quarato
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, via L. Pinto c/o OO.RR., 71100 Foggia, Italy.
authors.forename: Rosella
authors.initials: R
authors.lastname: Scrima
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, via L. Pinto c/o OO.RR., 71100 Foggia, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Ripoli
authors.affiliation: Laboratory of Pre-Clinical and Translational Research, IRCCS, Referral Cancer Center of Basilicata, Rionero in Vulture, PT, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Agriesti
authors.affiliation: Division of Gastroenterology and Hepatology, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland.
authors.forename: Darius
authors.initials: D
authors.lastname: Moradpour
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, via L. Pinto c/o OO.RR., 71100 Foggia, Italy. Electronic address: nazzareno.capitanio@unifg.it.
authors.forename: Nazzareno
authors.initials: N
authors.lastname: Capitanio
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, via L. Pinto c/o OO.RR., 71100 Foggia, Italy. Electronic address: claudia.piccoli@unifg.it.
authors.forename: Claudia
authors.initials: C
authors.lastname: Piccoli
mesh_terms: D000547:Amantadine / Q000494:pharmacology*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D000978:Antiparkinson Agents / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology*; D020013:Calcium Signaling / Q000187:drug effects; D002460:Cell Line; D045222:Electron Transport Chain Complex Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D016174:Hepacivirus / Q000187:drug effects* / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D006801:Humans; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism / Q000821:virology; D010085:Oxidative Phosphorylation / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects*; D020815:Polyproteins / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000819:agonists / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D000978:Antiparkinson Agents; D000998:Antiviral Agents; D045222:Electron Transport Chain Complex Proteins; D020815:Polyproteins; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D014764:Viral Proteins; D000547:Amantadine
keywords: Amantadine; Amantadine hydrochloride (PubChem CID: 64150); Calcium channels; DCF (CID: 64944); FCCP (CID: 3330); Hepatitis C virus; Inducible viral protein expression; Mitochondria; Oxidative stress; Rhod-1 (CID: 44140597); Ru 360 (CID: 16760639); TMRE (CID: 2762682); oligomycin (CID: 6450197); ruthenium red (CID: 44134938); tetracycline (CID: 54675776)