Thioridazine interacts with the membrane of mitochondria acquiring antioxidant activity toward apoptosis--potentially implicated mechanisms.
We evaluated the effects of the phenothiazine derivative thioridazine on mechanisms of mitochondria potentially implicated in apoptosis, such as those involving reactive oxygen species (ROS) and cytochrome c release, as well as the involvement of drug interaction with mitochondrial membrane in these effects. Within the 0 - 100 microM range thioridazine did not reduce the free radical 1,1-diphenyl-2-picryl-hydrazyl (DPPH) nor did it chelate iron. However, at 10 microM thioridazine showed important antioxidant activity on mitochondria, characterized by inhibition of accumulation of mitochondria-generated O2*-, assayed as lucigenin-derived chemiluminescence, inhibition of Fe2+/citrate-mediated lipid peroxidation of the mitochondrial membrane (LPO), assayed as malondialdehyde generation, and inhibition of Ca2+/t-butyl hydroperoxide (t-BOOH)-induced mitochondrial permeability transition (MPT)/protein-thiol oxidation, assayed as mitochondrial swelling. Thioridazine respectively increased and decreased the fluorescence responses of mitochondria labelled with 1-aniline-8-naphthalene sulfonate (ANS) and 1-(4-trimethylammonium phenyl)-6 phenyl 1,3,5-hexatriene (TMA-DPH). The inhibition of LPO and MPT onset correlated well with the inhibition of cytochrome c release from mitochondria. We conclude that thioridazine interacts with the inner membrane of mitochondria, more likely close to its surface, acquiring antioxidant activity toward processes with potential implications in apoptosis such as O2*- accumulation, as well as LPO, MPT and associated release of cytochrome c.
Rodrigues
Tiago
T
Department of Physics & Chemistry, Faculty of Pharmaceutical Sciences, University of São Paulo, 14040-903 Ribeirão Preto, São Paulo, Brazil.
Santos
Antonio C
AC
Pigoso
Acácio A
AA
Mingatto
Fábio E
FE
Uyemura
Sérgio A
SA
Curti
Carlos
C
D000817:Anilino Naphthalenesulfonates; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D014150:Antipsychotic Agents / Q000494:pharmacology*; D017209:Apoptosis; D002118:Calcium / Q000378:metabolism; D003574:Cytochrome c Group / Q000378:metabolism; D004161:Diphenylhexatriene / Q000031:analogs & derivatives*; D004560:Electricity; D005453:Fluorescence; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D010101:Oxygen Consumption / Q000187:drug effects; D010539:Permeability; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D013881:Thioridazine / Q000494:pharmacology*; D014268:Trifluoperazine / Q000494:pharmacology; D020122:tert-Butylhydroperoxide / Q000378:metabolism
D000817:Anilino Naphthalenesulfonates; D000975:Antioxidants; D014150:Antipsychotic Agents; D003574:Cytochrome c Group; D005456:Fluorescent Dyes; D017382:Reactive Oxygen Species; D004161:Diphenylhexatriene; D014268:Trifluoperazine; C027132:1-anilino-8-naphthalenesulfonate; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene; D020122:tert-Butylhydroperoxide; D013881:Thioridazine; D002118:Calcium