芍药苷的体外代谢性质  

Metabolic characteristics of paeoniflorin in vitro

作　　者：谭妍[1] 沈国林[1] 庄笑梅[1] 原梅[1] 李桦[1] 高月 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850 [2]放射与辐射医学研究所,北京100850

出　　处：《国际药学研究杂志》2013年第5期625-629,633,共6页Journal of International Pharmaceutical Research

基　　金：国家自然科学基金重点课题(81130067);国家“重大新药创制”科技重大专项(2012ZX09301003-001)

摘　　要：目的研究芍药苷在人和大鼠肝微粒体,以及大鼠原代肝细胞中的代谢特征,确定芍药苷的Ⅰ相代谢酶表型。方法将芍药苷与人、大鼠肝微粒体或大鼠肝细胞在37℃孵育,应用液相色谱-串联质谱(LC-MS/MS)法测定孵育液中的芍药苷含量,考察芍药苷的代谢稳定性。将芍药苷与重组人源细胞色素P450酶(CYP)中CYP1A2、2B6、2C8、2C9、2C19、2D6和3A4孵育,并在人肝微粒体中将芍药苷与各同工酶的特异性抑制剂共孵育,确定其代谢酶表型。结果在人和大鼠肝微粒体中,芍药苷均发生依赖于NADPH的Ⅰ相代谢消除,60 min的代谢率分别为10.74%和22.76%,半衰期T1/2分别为(88.87±5.32)和(48.47±1.18)min,经外推得到的肝脏清除率分别为(13.13±1.11)和(10.06±1.34)ml/(min·kg)。在加入葡萄糖醛酸转移酶辅酶(UDPGA)的人肝微粒体中,芍药苷不发生代谢转化。在大鼠原代肝细胞中,芍药苷的代谢转化率与肝微粒体相当,T1/2为(77.88±3.93)min。芍药苷是由多个CYP同工酶介导代谢的,CYP2C9、CYP3A4和CYP2C8是主要的代谢酶,用整体归一化法评价其代谢贡献率分别为22.06%、52.47%和25.47%。结论芍药苷在肝微粒体和肝细胞中主要发生多个CYP酶介导的Ⅰ相代谢,主要的代谢表型是CYP2C9、CYP3A4和CYP2C8。Objective To investigate the metabolic characteristics of paeoniflorin in liver microsomes and primary cultured hepatocytes in vitro, and to identify the metabolic phenotyping of phase I metabolism. Methods Paeoniflorin was incubated at 37℃ with human and rat liver microsomes in the presence or absence of NADPH or UDGPA, and also incubated with rat primary cultured hepatocytes. The concentrations of paeoniflorin in the above incubation systems were determined by a LC-MS/MS method to evaluate the metabolic stability of paeoniflorin. The eytoehrome P450 （CYP） phenotyping of paeoniflorin was identified using a panel of rCYP isoforms （ CYP1 A2,2B6,2C8,2C9,2C19,2I）6 and 3A4 ） and specific inhibitors of CYP isoforms in human liver microsomal incubation system. Results In human and rat liver microsomes, paeoniflorin could be metabolically eliminated in the presence of NADPH. The metabolic rates in 60 rain were 10. 74% and 22. 76% and the T1/2 of paeoniflorin was （88. 87 ±5.32） rain and （48.47 ± 1.18） rain for human and rat, respectively. The extrapolated hepatic clearance was （ 13.13 ± 1.11 ） and （ 10. 06 ± 1.34） ml/（ min . kg）. In the presence of UDPGA, the cosubstrate of UDP glucuronosyhransferase, paeoniflorin was not metabolized in the human liver microsomes. In the rat primary cultured hepatocytes, paeoniflorin showed a comparable metabolic profile as in the rat liver microsomes with the T1/2 of （77.88 ±3.93） rain. The results of CYP phenotyping indicated that CYP2C8, 2C9 and 3A4 were involved in the metabolism of paeoniflorin. Their individual contributions were assessed using the method of the total normalized rate to be 22.06% , 52. 47% 25.47%, respectively. Conclusion Paeoniflorin is mainly metabolized by a number of CYP isoenzymes in liver microsomes and hepatocytes. CYP2C9, 3A4 and 2C8 are the major metabolic enzymes responsible for the metabolism of paeoniflorin.

关 键 词：芍药苷 细胞色素P450酶 肝微粒体 肝细胞 代谢 

分 类 号：R285[医药卫生—中药学]