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Disruption of Mitochondrial Calcium Homeostasis after Chronic α-Naphthylisothiocyanate Administration: Relevance for Cholestasis
  1. Anabela P. Rolo,
  2. Paulo J. Oliveira,
  3. Raquel Seiça,
  4. Maria S. Santos,
  5. António J. Moreno,
  6. Carlos M. Palmeira
  1. From the Department of Zoology (A.P.R., P.J.O., M.S.S., A.J.M., and C.M.P.) University of Coimbra, Portugal
  2. Faculty of Medicine (R.S.), Center for Neurosciences and Cell Biology of Coimbra, University of Coimbra, Portugal.
  1. Address correspondence to: Carlos M. Palmeira, PhD, Department of Zoology, Center for Neurosciences and Cell Biology of Coimbra, University of Coimbra, 3004-517 Coimbra, Portugal. Email: palmeira{at}
  2. A.P.R. and P.J.O. are recipients of PRAXIS Grants XXI/BD/21454/99 and XXI/21494/99, respectively, from the Fundação para a Ciência e Tecnologia (FCT), Lisbon, Portugal.


Background Hepatocyte dysfunction caused by impaired mitochondrial function has been pointed out as a probable leading cause of cholestatic liver injury. The aim of this study was to evaluate liver mitochondrial bioenergetics that followed repeated in vivo administration of α-naphthylisothiocyanate, a known cholestatic agent.

Methods Serum markers of liver injury and endogenous adenine nucleotides were measured in α-naphthylisothiocyanate-treated rats (intraperitoneally, 100 mg/Kg/wkx6 wk). Changes in membrane potential, mitochondrial respiration, as well as alterations in mitochondrial calcium homeostasis were monitored.

Results In rats injected with α-naphthylisothiocyanate, liver injury with cholestasis developed within 48 hours, as indicated by both serum enzyme activities and total bilirubin concentration. However, 1 week after the last injection, serum enzyme activity returned to control levels. In addition, after chronic α-naphthylisothiocyanate administration, no alterations in mitochondrial respiratory function and membrane potential were observed. Associated with these parameters, mitochondria from treated animals exhibited increased susceptibility to disruption of mitochondrial calcium homeostasis by calcium phosphate and by bile acids, which was probably caused by induction of permeability transition pore.

Conclusions Our data suggest that chronic cholestasis in rats leads to impaired mitochondrial function due to the disruption of mitochondrial calcium homeostasis. The initiating event is the induction of a cyclosporine A-sensitive release of calcium. This event may be an important determinant of the progression of cholestatic liver injury and associated liver cirrhosis. In addition, in the present study we observed that impairment of mitochondrial function is potentiated by chenodeoxycholate, a bile acid that is known to be toxic. Ursodeoxycholate (the β- epimer of chenodeoxycholate) is approved for the treatment of chronic cholestatic liver disease. Interestingly, we show that the susceptibility to the cyclosporine A-sensitive release of calcium was increased by the combination of both bile acids. These results indicate that the reported improvement of biochemical parameters in cholestatic patients treated with ursodeoxycholate would not prevent the associated mitochondrial dysfunction. This may explain the progression of the histological stage and the maintenance of symptoms during cholestasis.

Key Words
  • mitochondria
  • permeability transition
  • membrane potential
  • respiration
  • calcium
  • bile acids

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