大鼠离体原代肝细胞中布洛芬代谢的研究

Metabolism of Ibuprofen in Primarily Cultured Hepatocytes of Rats

目的:研究布洛芬在大鼠原代培养肝细胞中的代谢情况。方法:建立大鼠肝细胞的分离及原代培养方法,考察肝细胞对布洛芬的代谢情况;采用2步灌流技术并运用胶原酶消化法,分离肝细胞并分别与2.50、5.00、20.00μg·mL-1浓度布洛芬进行悬浮共培养,在不同时间点取样,应用高效液相色谱法测定布洛芬的浓度并计算其它药动学参数。结果:240min内肝细胞活率>80%,能满足代谢要求;布洛芬低、中、高3种浓度药动学数据Ke为(0.0064±0.0011)、(0.0086±0.009)、(0.0091±0.0015)min-1,t1/2为(110.0562±17.16)、(81.3873±9.2815)、(77.9516±12.1355)min;AUC为(272.9287±35.58)、(448.9899±23.0295)、(2157.6777±318.8730)μg·mL·min-1;CLs为(0.0093±0.0011)、(0.0112±0.0005)、(0.0094±0.0014)mL·min-1。结论:利用原代肝细胞培养技术对布洛芬代谢进行研究,可为临床优化治疗、合理用药等方面提供有价值的实验数据。

OBJECTIVE： To study the metabolism of ibuprofen in primarily cultured hepatocytes of rats. METHODS： The method for the isolation and primary culture of rats＇ hepatocytes was establisehed and the metabolism of ibuprofen in isolated hepatocytes was investigated. Hepatocytes were isolated from rats using two step perfusion and collagenase digestion procedure and co-cultured with different concentrations of ibuprofen （2.50, 5.00, and 20.00 μg·mL^-1, respectively） . Samples were taken at different time point for the HPLC determination of concentration of ibuprofen in hepatocyte suspension and computation of its pharmacokinetic parameters. RESULTS： The metabolism of ibuprofen could be achieved if hepatocytes activity was more than 80% in 240 min. The pharmacokinetic parameters of ibuprofen at low, middle and high concentrations were as follows： Ke： （0.006±0.0011）, （0.0086±0.009）, and （0.0091±0.0015） min^-1, respectively; t1/2（110.0562±17.16）, （81.3873±9.2815）, and （77.9516±12.1355） min, respectively; AUC： （272.9287±35.58）, （448.9899±23.0295）, and （2 157.6777±318.8730）μg·mL·min^-1, respectively. CLs： （0.0093±0.0011）, （0.0112±0.0005）, and （0.0094±0.0014） mL·min^-1 respectively. CONCLUSION： Applying primary hepatocyte culture technology in the study of metabolism of ibuprofen cau provide valuable experiment data for clinical optimal treatment and rational use of drugs etc.