Richter P, Kauffman F C, Zaleski J
Department of Pharmacology and Toxicology, Rutgers University, Piscataway, New Jersey 08854-5930.
Toxicol Lett. 1994 Oct;74(1):79-90. doi: 10.1016/0378-4274(94)90076-0.
This study shows that 1-hydroxybenzo[a]pyrene glucuronide and 1-hydroxybenzo[a]pyrene sulfate are formed in isolated rat hepatocytes. Formation of these conjugates by hepatocytes incubated with 1-acetoxy-[G-3H]benzo[a]pyrene (100 microM) as a source of intracellular 1-hydroxy-[G-3H]benzo[a]pyrene was documented by comparison of the spectra of metabolites separated by HPLC with the spectra of 1-hydroxybenzo[a]pyrene glucuronide and 1-hydroxybenzo[a]pyrene sulfate standards. The rates of 1-hydroxybenzo[a]pyrene glucuronidation and sulfation were 7.72 +/- 1.03 and 0.68 +/- 0.02 nmol x mg dry wt.-1 x 30 min-1, respectively. The rate of 1-hydroxybenzo[a]pyrene glucuronide production by intact cells corresponded well with the total activity of UDP-glucuronosyltransferase(s) determined in permeabilized hepatocytes. Cryopreserved hepatocytes fully retained a high capacity to glucuronidate the benzo[a]pyrene phenol.
本研究表明，在分离的大鼠肝细胞中会形成1-羟基苯并[a]芘葡萄糖醛酸苷和1-羟基苯并[a]芘硫酸盐。通过将经高效液相色谱法分离的代谢物光谱与1-羟基苯并[a]芘葡萄糖醛酸苷和1-羟基苯并[a]芘硫酸盐标准品的光谱进行比较，证明了以1-乙酰氧基-[G-3H]苯并[a]芘（100微摩尔）作为细胞内1-羟基-[G-3H]苯并[a]芘来源时，肝细胞会形成这些共轭物。1-羟基苯并[a]芘葡萄糖醛酸化和硫酸化的速率分别为7.72±1.03和0.68±0.02纳摩尔×毫克干重-1×30分钟-1。完整细胞产生1-羟基苯并[a]芘葡萄糖醛酸苷的速率与在透化肝细胞中测定的尿苷二磷酸葡萄糖醛酸基转移酶的总活性相当。冷冻保存的肝细胞完全保留了对苯并[a]芘酚进行葡萄糖醛酸化的高能力。
