Acrylonitrile-induced reversible inhibition of uridine uptake by isolated rat intestinal epithelial cells

Isolated rat intestinal epithelial cells (IEC) were used to investigate the mechanism(s) of acrylonitrile (VCN)-induced gastro-intestinal damage. The isolated cells were 93% structurally intact for 60 min, as indicated by trypan blue exclusion. Uridine uptake by isolated IEC was linear from 6-20 min, after which a steady state was reached for up to 40 min. Exposure of isolated IEC to various concentrations of VCN reduced the ability of the cells to take up uridine in a concentration-dependent manner. A concentration of 82 mum VCN inhibited the [(3)H]uridine uptake of the cells by 50% (IU(50)). A time-course study indicated that the maximal inhibition of uridine uptake occurred at 15 min after exposure to VCN. The VCN-induced inhibition of uridine uptake was found to be reversible. IEC exposed to two sublethal doses of VCN (41 and 82 mum) for 15 min regained normal uridine uptake activity within 50 min after removal of VCN. The present study provides a sensitive approach for the detection and evaluation of cytotoxic risk of sublethal doses of the gastro-intestinal toxin VCN using IEC as target cells. The observed in vitro cytotoxicity of sublethal doses of VCN will be used to investigate further the mechanism of VCN-induced gastro-intestinal damage.