title: Real-time concurrent monitoring of apoptosis, cytosolic calcium, and mitochondria permeability transition for hypermulticolor high-content screening of drug-induced mitochondrial dysfunction-mediated hepatotoxicity.
abstract: A quantitative high-content screening (HCS) was suggested for the real-time monitoring of drug-induced mitochondrial dysfunction-mediated hepatotoxicity. This HCS is very advantageous in that it allows simultaneous observation of drug-induced activations of hepatotoxic pathways using hypermulticolor cellular imaging. The mitochondrial permeability transition (MPT), cytosolic calcium, and caspase-3 were selected as functional markers to verify drug-induced hepatotoxicity and were concurrently monitored in HepG2 cells in a real-time manner. Nefazodone, tolcapone, and troglitazone caused mitochondrial dysfunction and subsequent apoptotic HepG2 cell death in addition to marked cytosolic calcium increase. On the other hand, extrinsic pathway-mediated apoptotic cell death was monitored when HepG2 cells were treated with piroxicam. It was found that piroxicam-treated HepG2 cells showed apoptotic cell death without the MPT formation, while a cytosolic calcium increase was clearly observed. This finding was confirmed by the caspase-8 inhibition assay. These results demonstrated the unique potential of real-time hypermulticolor HCS to screen hepatotoxic drugs at the in vitro stage rather than the later in vivo stage based on an animal model and to ultimately reduce the probability of drug failure.
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Jae-Ah
authors.initials: JA
authors.lastname: Kim
authors.forename: Eunyoung
authors.initials: E
authors.lastname: Han
authors.forename: Chae-Jung
authors.initials: CJ
authors.lastname: Eun
authors.forename: Yu Kyung
authors.initials: YK
authors.lastname: Tak
authors.forename: Joon Myong
authors.initials: JM
authors.lastname: Song
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D001577:Benzophenones / Q000633:toxicity*; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D002839:Chromans / Q000633:toxicity*; D056945:Hep G2 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007093:Imidazoles / Q000633:toxicity*; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism; D033681:Mitochondrial Membrane Transport Proteins / Q000187:drug effects / Q000378:metabolism; D000083162:Mitochondrial Permeability Transition Pore; D009596:Nitrophenols / Q000633:toxicity*; D010879:Piperazines; D045162:Thiazolidinediones / Q000633:toxicity*; D000077867:Tolcapone; D014230:Triazoles / Q000633:toxicity*; D000077288:Troglitazone
chemical_list: D001577:Benzophenones; D002839:Chromans; D007093:Imidazoles; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D009596:Nitrophenols; D010879:Piperazines; D045162:Thiazolidinediones; D014230:Triazoles; C039686:piroximone; C051752:nefazodone; D000077867:Tolcapone; D053148:Caspase 3; D000077288:Troglitazone; D002118:Calcium