NORMAL AND RETROGRADE PERFUSION TO PROBE THE ZONAL DISTRIBUTION OF SULFATION AND GLUCURONIDATION ACTIVITIES OF HARMOL IN THE PERFUSED-RAT-LIVER PREPARATION

Abstract

The role of zonal distributions of metabolic activities (sulfation and glucuronidation) in the liver on the kinetics of harmol conjugation was investigated. A computer simulation approach was adopted to better understand the effects of distributions of these conjugation activities in competition for a common drug substrate. Several distributions of the sulfation and glucuronidation systems were defined with respect to the flow path of blood; the conjugation activities along the flow path were in turn translated as time elapsed after entry of the substrate via blood into the organ. Realistic values of Km and Vmax for the sulfation and glucuronidation systems were assigned in the simulations. Directional flow, i.e., normal vs. retrograde delivery of substrate was used as an additional variable. When the center of distribution of the sulfation system was anterior to that for glucuronidation, the steady-state hepatic extraction ratio of harmol (E) would increase, whereas the ratio of the steady-state rates of formation of harmol sulfate to harmol glucuronide (S/G ratio) would decrease when harmol was presented in a reversed direction (via retrograde perfusion), as compared with normal directional flow to the liver. The converse was true for an anterior distribution of the glucuronidation system; E would decrease, whereas S/G would increase with retrograde perfusion. To evaluate such zonal differences experimentally, perfusion studies were conducted in livers of male Wistar rats. A constant concentration of harmol (50 .mu.M) was delivered under constant hepatic flow rate (10 ml/min per liver) by normal and retrograde perfusions to the same rat liver preparation. The steady-state hepatic E was higher (P < 0.005) during retrograde perfusion than during normal perfusion, whereas the S/G ratio was significantly decreased (P < 0.0005). An apparent intercellular difference in the distribution of the 2 conjugating systems is consistent with the view of the center of distribution for the sulfation system being anterior to the center of distribution for the glucuronidation system along the normal flow path of blood in the liver.