体外探针法评价五味子和南五味子提取物对人肝微粒体CYP3A4酶活性的影响

Evaluation of the Effects of Wuweizi and Nan Wuweizi Extract on Human Liver Microsomal CYP3A4 Enzyme Activity by in Vitro Probe Method

目的 通过体外药物探针实验探讨五味子和南五味子提取物和主要木脂素成分对细胞色素P450 3A4(cytochrome P450 3A4,CYP3A4)酶活性的影响。方法 选取睾酮为药物探针,以对乙酰氨基酚为内标,建立高效液相色谱-串联质谱法检测人肝微粒体中代谢物6β-羟基睾酮含量方法。结果 6β-羟基睾酮的线性范围为15.0~960μg/L,相关系数为0.999 9;相对回收率为102.1%~105.7%,其相对标准偏差均小于11%。五味子和南五味子醇提物和水提物对CYP3A4酶活性均有抑制作用,五味子醇提物和水提物的抑制能力强于南五味子,此外二者的醇提物的抑制程度强于其水提物。木脂素类成分对CYP3A4酶活性的抑制作用强弱程度依次为:五味子酯甲、五味子乙素、五味子甲素、五味子醇甲。结论 五味子和南五味子提取物对CYP3A4酶活性的抑制作用差异较显著(P<0.05)。临床上二者混用现象偶见发生,如果与CYP3A4酶代谢特异性高的药物联合使用时,可能会因为代谢性的相互作用引起一些不良后果。

Objective The aim was to evaluate the effects of Wuweizi and Nan Wuweizi extract on the enzyme activities of human hepatic cytochrome P450 3A4(CYP3A4) by in vitro probe.Methods Microsomal CYP3A4 activity was determined by measuring the amount of 6β-hydroxytestosterone by liquid chromatography-tandem mass spectrometry combined with a drug probe of testosterone.Results The linear range of 6β-hydroxytestosterone was 15.0~960 μg/L and the correlation coefficient was 0.999 9.The relative recovery was 102.1%~105.7% and the value of relative standard deviation was less than 11%.The activity of the CYP3A4 enzyme was inhibited by the alcohol and aqueous extracts of Wuweizi and Nan Wuweizi.The inhibitory abilities of the alcohol and aqueous extracts of Wuweizi were stronger than those of Nan Wuweizi.In addition,the inhibitory degree of alcohol extracts was stronger than that of water extracts.The inhibitory effect of lignans on the activity of CYP3A4 enzyme was in the order:schisantherin A,schizandrin B,schisandrin A,schizandra A.Conclusion The inhibitory effects of Wuweizi and Nan Wuweizi extracts on CYP3A4 enzyme activity were significantly different(P< 0.05).Clinically,the phenomenon of their mixed-use occurred occasionally.If they were in combination with the drugs whose metabolism involves CYP3A4 specificity,some adverse consequences would be caused by metabolic interaction.