[Release of cytochrome C with the interaction of bilirubin, amyloid beta-peptide and glycochenodeoxycholate from isolated mitochondria].
The pathogenesis of cholestasis, bilirubin encephalopathy, and Alzheimer's disease appears to result from accumulation of diverse cytotoxic agents, which in turn may cause apoptotic cell death. In addition, mitochondria has lately been considered as a central executioner of programmed cell death, through the release of caspase activating factors. The aims of this study were to: (a) investigate mitochondrial perturbation during incubation of isolated mitochondria with unconjugated bilirubin (Bb), amyloid beta-peptide (A beta), and glycochenodeoxycholate (GCDC); (b) characterize membrane perturbation in isolated mitochondria induced by each toxic agent, and determine whether the mitochondrial permeabilization is required for cytochrome c redistribution. Mitochondria were isolated from rat liver and brain. Swelling and cytochrome c release were evaluated by spectrophotometry and western blot, respectively. The results showed that Bb as well as A beta and GCDC act directly at the mitochondrial level causing increased organelle volume, permeabilization, as well as cytochrome c release from the intermembrane space in a dose-dependent manner (P < 0.01). Moreover, cyclosporine A inhibited mitochondrial permeability, particularly after Bb- and GCDC-induced swelling (P < 0.01). Cytochrome c efflux was invariably prevented by cyclosporine A (P < 0.05). In conclusion, the results indicate that Bb-, A beta-, and GCDC-induced toxicity, culminating in apoptosis, may result from enhanced mitochondrial permeability, followed by cytochrome c efflux, which can be explained at least in part by the megapore opening.
Solá
Susana
S
Centro de Patogénese Molecular, Faculdade de Farmácia da Universidade de Lisboa.
Diógenes
Maria José
MJ
Brites
Dora
D
Rodrigues
Cecília M P
CM
D016229:Amyloid beta-Peptides / Q000502:physiology*; D000818:Animals; D001663:Bilirubin / Q000502:physiology*; D003574:Cytochrome c Group / Q000378:metabolism*; D005999:Glycochenodeoxycholic Acid / Q000502:physiology*; D008928:Mitochondria / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D016229:Amyloid beta-Peptides; D003574:Cytochrome c Group; D005999:Glycochenodeoxycholic Acid; D001663:Bilirubin