Study on glutathione S-transferase inhibition assay by triclabendazole. III: nematodirus parasite and sheep liver tissue

ABSTRACT

The most important and widely prevalent nematodes of sheep are the trichostrongyle group parasites, including nematodirus parasite. Accidental infection of man by nematodirus has been reported in Iran. Glutathione S-Transferase enzymes [GSTs] are detoxification enzymes in parasites such as nematodirus. Therefore, GST enzymes of these parasites could be a target for evaluation of drugs effect as triclabendazole [C[14]H[9]CL[3]N[[2]OS]. For this reason, GST enzymes were purified from nematodirus parasite and sheep liver tissue by glutathione affinity chromatography and prepared their SDS-PAGE banding pattern for GST fraction separation. GST enzymes specific activity levels are also assayed in the whole extract and purified solutions with reduced glutathione [GSH] and l-chloro-2, 4-dinitrobenzen [CDNB] secondary substrate. Finally, GST inhibition assay was investigated in the solutions by powder and bolus forms of triclabendazole. The level of GST specific activity in purified solutions was detected 9.86 micromol / min/ mg protein for nematodirus parasite and 37.84 micromol/ min/ mg protein for liver tissue. Comparison of the effect of powder and bolus of triclabendazole on solutions revealed inhibition concentration [IC[50]] 5.54 and 6.01 microg/ml for nematodirus GST and 8.65 and 9.70 microg/ml for liver tissue GST, respectively. These findings revealed the possibility of isolation and inhibition of nematodirus GST by triclabendazole, and more tolerance of liver tissue than parasite against this drug in vitro situation