【摘要】 目的:比较大鼠肝细胞在单层贴壁、胶原凝胶包埋和聚球体三种培养方式下药物代谢酶的活性及β-萘黄酮(BNF)对酶的诱导作用,考察组织化培养大鼠肝细胞的药物代谢能力。方法:大鼠肝细胞经三种培养方式培养一定时间后,分别加入非那西丁和7-羟基香豆素(7-HC),高效液相色谱法测定代谢产物对乙酰氨基酚、葡萄糖化7-羟基香豆素(7-HC GLUC)和硫酸化7羟-基香豆素(7-HC SULF)的浓度,分别用于表征Ⅰ、Ⅱ相药物代谢酶的活性。以BNF作为药物代谢酶的诱导剂,分别考察它对Ⅰ、Ⅱ相药物代谢酶的诱导情况。结果:单层贴壁培养肝细胞Ⅰ相药物代谢酶活性在第5天已检测不到,Ⅱ相药物代谢酶的活性在第7天已低于检测下限。在胶原凝胶包埋和聚球体两种培养方式下,大鼠肝细胞的Ⅰ和Ⅱ相酶活性在第7天还能保持初始值的32%~50%,且肝细胞聚球体的对乙酰氨基酚生成速率到第7天仍维持第1天的96%。肝细胞培养1~3 d,肝细胞聚球体培养Ⅰ相药物代谢酶的活性比单层贴壁培养高2.7~3.9倍(P<0.01,P<0.05)。BNF处理后,三种培养模型的Ⅰ相酶的活性都有2.5倍左右的提高(P<0.05),而Ⅱ相酶的活性增加并不明显。结论:在保持药物代谢酶活性方面,肝细胞的胶原凝胶包埋和聚球体培养方式比单层贴壁培养方式更优,可能成为研究药物代谢和药物相互作用更合适的体外模型。更多还原

【Abstract】 Objective: To investigate phase Ⅰ and phase Ⅱ enzyme activities in drug metabolism with tissue-like cultures of rat hepatocytes.Methods: The gel entrapment and spheroid culture of hepatocytes were used as tissue-like cultures and the monolayer culture was used as a control.The metabolism of phenacetin and 7-hydroxycoumarin（7-HC） was evaluated as the activities of phase Ⅰand phase Ⅱenzymes after incubated in medium for a period of time.The metabolites were assayed by HPLC.The hepatocytes were exposed to β-naphthoflavone(BNF,50 μmol·L^-1) before the phaseⅠand phase Ⅱenzyme activities were analyzed.Results: In monolayer culture,phase Ⅰ parameters decreased quickly and did not detected at d 5,and the phase Ⅱ enzyme activities were not detected at d 7.In other two models of tissue-like cultures,the activities of phase Ⅰ and phase Ⅱ enzyme maintained at 32%⁵0% of the initial value at d 7.Paracetanol formation rates in spheroid culture maintained at 96% of that at d 1.The phase Ⅰ enzyme activities of the spheroid culture were maintained from d 1 to d 3 at a level of 2.7³.9-fold higher than the monolayer culture.After exposure to BNF the activities on phaseⅠenzyme increased by about 2.5-fold（P<0.05） in all three culture models,while the increase in phase Ⅱ enzyme was not significant.Conclusion: The gel entrapment culture and spheroid culture are superior to the monolayer culture in maintenance of drug metabolic enzyme activities.更多还原