Species- and congener-differences in microcystin-LR and -RR GSH conjugation in human, rat, and mouse hepatic cytosol.

The accepted pathway for MC biotransformation is GSH conjugation, occurring either spontaneously or catalyzed by GST. In the present work, the already available information on human MC metabolism have been expanded and the capacity of human GST to conjugate MC-LR has been confirmed in human liver cytosol. At physiological GSH content the spontaneous reaction predominated on the enzymatic one; the prevalence of the enzymatic reaction occurred following GSH depletion, and the shift was detectable at higher GSH levels, the lower was MC concentration. However, at low MC-LR concentrations (≤10μM), representative of repeated oral exposure, the relevance of the enzymatic reaction became predominant at GSH concentration between 1 and 2mM. MC-LR conjugate was detectable at ≥0.5mM GSH, whereas, with 10μM MC-RR detectable levels of conjugate were observed at 0.05mM GSH, a 10-fold lower concentration. Overall, our data indicate that MC-RR is more efficiently conjugated than MC-LR, especially at low concentrations. Cytosol samples from rat and mouse were used to characterize GSH conjugation of MC-LR and MC-RR, and to check for possible species differences. At physiological GSH content, in both rodent species the enzymatic reaction accounted for half of the total conjugate formation, reducing the impact of spontaneous reaction with respect to human. Rat and mouse GST showed similar MC-LR and-RR GSH conjugation, but a two-fold higher catalytic efficiency than human sample. This is mainly due to higher affinity for the substrate, with Kmapp values being an order of magnitude lower in the animal models than in human liver cytosol. More pronounced differences in the metabolism of the two variants were evidenced in rodents than in humans.