THE EFFECTS OF CYCLOSPORINE-A (CSA) ON HEPATIC-MICROSOMAL DRUG-METABOLISM IN THE RAT

Abstract

The effects of cyclosporin A (CsA), a powerful immunosuppressant, on the hepatic microsomal mixed funciton oxidase (MFO) system was studied in male rats. Difference spectroscopy studies indicated that CsA binds to cytochrome P-450 producing a type I spectral change. To investigate potential interactions with the MFO system, CsA was administered orally at doses of either 25 mg/kg or 50 mg/kg once daily for 9 days. Various metabolic parameters were examined, including: levels of microsomal protein, cytochrome P-450, and cytochrome b5, NADPH-cytochrome c reductase activity, N-demethylation of ethylmorphine (ETM), and p-hydroxylation of aniline (ANL). Rats treated with 50 mg/kg showed a 25% greater decrease over controls in all parameters examined except microsomal protein and cytochrome b5 levels. Rats treated with 25 mg/kg showed a 28% or greater decrease in all parameters except microsomal protein, cytochrome b5, and cytochrome P-450. Kinetic studies of ETM-N-demethylase and ANL-hydroxylase activities were conducted either with microsomes prepared from CsA-treated animals (50 mg/kg/day for 5 days) or with pooled microsomes prepared from untreated animals to which CsA was added directly. Enzyme reaction velocities were measured and apparent KM and apparent Vmax were determined. Studies with CsA-treated animals revealed a 57% decrease in both KM and Vmax for ETM-N-demethylase, and a 46% decrease in KM and 32% decrease in Vmax for ANL-hydroxylase. Studies involving direct addition of CsA to microsomes at final concentrations of 0.01 mM and 1.10 mM revealed no significant changes in apparent KM or Vmax for either enzyme. For the majority of parameters examined, an inhibitory effect on the heaptic microsomal MFO system of CsA-treated animals is evident, but microsomal metabolism appears to be unaffected by direct in vitro addition of CsA.