Effect of diet and β-naphthoflavone on hepatic and renal glutathione S-transferase isoenzymes in carp (Cyprinus carpio)

Abstract

Glutathione S-transferase (GST) isoenzymes were isolated from liver and kidney of carp (Cyprinus carpio) by glutathione affinity chromatography and chromatofocusing. Ten hepatic and eight renal catalytically active isoenzymes were identified. GST subunits from purified isoenzymes were further separated by high-performance liquid chromatography (HPLC) and were used as standards for the experimental sample analysis. Experimental samples came from carp that were fed fish meal (standard diet) or soybean bas ed diets for one year, injected or not with β-naphthoflavone (BNF; i.p. injection of 50 mg kg-1). HPLC did not allow us to identify precisely the GST isoenzyme pattern in experimental carp. However, GSTs could be pooled in three categories: homodimeric, heterodimeric and unidentified GST subunits. On this basis, the effect of diet and BNF on the GST isoenzyme pattern was investigated. The homodimer/heterodimer ratio was decreased in liver of carp fed a standard diet and in kidney of both dietary groups. BNF increased the total specific GST activity in liver and kidney. However, the GST isoenzyme pattern was not modified in carp fed the standard diet while tissue specific modifications occured in carp fed the soybean diet. BNF decreased the homodimer/heterodimer ratio in liver and increased it in kidney. Abbreviations: BNF - β-naphthoflavone; CDNB - 1-chloro-2,4-dinitrobenzene; GSH - reduced glutathione; GST - glutathione S-transferase; HPLC - high-performance liquid chromatography; SDS-PAGE - SDS-polyacrylamide gel electrophoresis.