Characterization of the Hepatic Uptake of l-Anilino-8-naphthalene sulfonate(ANS) by Isolated Rat Hepatocytes-Is Serum Protein Essential for Hepatic Uptake of ANS in the Liver?-

유리간세포를 사용한 ANS의 간내 이행에 관한 연구-ANS의 간내 이행과정에 단백질 매개 기구가 존재하는가?-

The hepatic uptake of an anionic fluorescence probe, l-anilino-8-naphthalene sulfonate (ANS) was characterized using isolated rat hepatocytes. The initial uptake rate of ANS by isolated hepatocytes was determined. The uptake process of ANS was fitted well to the Michaelis-Menten equation with a saturable component. The $V_{max}$ and $K_m$ values were $2.9{\pm}0.1\;nmol/min/mg$ protein and $29.1{\pm}3.2\;{\mu}M$, respectively. The uptake clearance $(CL_{up})$ based on the ratio of $V_{max}$ to $K_m$ was 11.7 ml/min/g liver, revealing the good coincidence with that assessed from the analysis of the plasma disappearance curve in previous report. Furthermore, the effect of serum protein on the hepatic uptake of ANS into isolated hepatocytes was investigated. The permeability clearances $(PS_{inf})$ of ANS uptake were much higher than those predicted based on the unbound fractions in the presence of serum. These suggested that the hepatic uptake of extensively serum protein-bound ANS is mediated not only by the unbound form of ligand but also by the serum protein-mediated uptake mechanism.