氮烯乙茶在成年大鼠肝细胞中的生物转化

目的:建立成年大鼠肝细胞分离培养方法,探讨抗癌新药氮烯乙茶在肝细胞内生物转化。方法:用肝细胞活率、乳酸脱氢酶、能荷值变化观察肝细胞悬浮培养方法的稳定性,以高效液相色谱法测定肝细胞悬液中氮烯乙茶及其代谢产物的浓度。结果:氮烯乙茶在大鼠肝细胞内生成代谢产物7-乙基-8-氨基茶碱(7-ethyl-8-aminotheophylline,EAT) ,其最大反应速度Vm 为12-80 nmol·min^-1/106 cells,米氏常数Km 为640-9 μmol·L^-1 ,氮烯乙茶的固有清除率CLint为20-0 μL·min^-1/106 cells。结论:肝细胞培养的方法是考察药物生物转化的较好体外模型,同时氮烯乙茶具有降低肝细胞能荷值的作用。

BIOTRANSFORMATION OF ETHEOFAZINE IN ADULT RAT HEPATOCYTE SUSPENSIONS

AIM: To establish a method for adult rat hepatocyte preparation and incubation and study the biotransformation of etheofazine, a new anticancer drug, in adult rat hepatocyte suspensions. METHODS: The concentration of etheofazine and its metabolite in hepatocyte suspensions was determined by HPLC method. In addition, the viability of adult rat hepatocyte was judged by the trypan blue exclusion test. Lactate dehydrogenase release and adenine nucleotide level were assessed. RESULTS: The metabolite in hepatocytes is 7 ethyl 8 aminotheophylline(EAT) and the result is in consistence with result obtained in isolated perfused rat liver.V_max=12 80 nmol·min^-1/10⁶ cells,Km=640.9 μmol·L^-1 , liver intrinsic clearance CL_int=20.0μL·min^-1/10⁶cells. CONCLUSION: Hepatocyte culture is shown to be a good in vitro model for drug biotransformation study and etheofazine can decrease the cellular energy charge in hepatocyte suspensions.