Paraquat resistance associated with reduced NADPH reductase in an energy-dependent paraquat-accumulating cell line

Abstract

Despite intensive investigation into paraquat toxicity, neither the final cytotoxic mechanism nor a clinically useful antidote has been discovered. In vitro screening of potential antidotes that act by blocking paraquat uptake requires a cell line that accumulates paraquat by an energy-dependent mechanism. We screened various lymphoblastoid cell lines until we found a line accumulating paraquat by an energy-dependent mechanism. During study of this cell line, a marked resistance to paraquat developed in a clone. The resistance was associated with a reduction in NADPH reductase activity, confirming the original report (using microsomal preparations) that intracellular reduction of paraquat occurs primarily by this enzyme. One-half of the NADPH-P450 reductase activity, as well as one-half of the NADPH-dependent paraquat-inducible superoxide production, was decreased. This suggests that the decrease is secondary to a genetic alteration in one of the genes encoding for the enzyme. Other antioxidant enzymes and proteins were not affected. Despite the loss of only 50% of the activity, the relative resistance to paraquat exceeded previous reports involving marked increases in antioxidant enzymes. Most exogenous enhancers or inhibitors alter the activity of more than one enzyme, thereby making selective changes in any one enzyme difficult. Thus, this cell line will be useful for studying other toxins where the involvement of NADPH reductase is suspected, but not proven.