一种微量肝组织体外孵育进行cyp450酶活性检测的方法

Method for minim hepatic tissue in vitro incubation and detecting CYP450 enzymatic activity

Abstract

一种微量肝组织体外孵育进行CYP450酶活性检测的方法,其特征在于:临床用肝微量穿刺法取0.1~0.2g微量肝组织,或动物试验处死小鼠取出肝脏,建立微量肝组织体外孵育体系,再用单探针药物进行液相色谱串联质潽法检测探针底物和相应代谢产物浓度,以它们的代谢比,求出P450酶的活性。本发明的优点是:1.微量:只需要0.1~0.2g的微量肝组织就可以进行细胞色素P450酶活性的检测,不仅可用于动物实验,而且可用于临床微量肝组织肝药酶活性检查;2.省时简便:省去了常规方法中需设备昂贵的超速离心,不仅大大节约了检测的成本和时间,而且更好地模拟了肝酶代谢环境;3.确立了可以评价酶代谢活性的指标:代谢比。
Disclosed is a method for detecting CYP450 enzymatic activity by means of micro-liver tissue incubation in vitro, which is characterized in that: a liver trace puncturing method used in clinic takes micro-liver tissues of 0.1 to 0.2g, or kills a mouse to take out the liver in an animal experiment, and builds a micro-liver tissue in vitro incubation system, then uses a one-probe medicine to perform the liquid phase chromatography tandem mass spectrometry for detecting probe substrates and concentration of corresponding metabolites, and obtains the activity of P450 enzyme according to metabolite ratios thereof. The method has the advantages of: 1. micro scale: only micro-liver tissues of 0.1 to 0.2g are needed for detecting cytochrome P450enzyme activity, and can be used for animal experiment and checking clinic micro-liver tissue liver drug enzyme activity; 2. time saving and convenience: costly ultracentrifugation equipment used in the conventional method is saved, costs and time for detection are greatly saved, and liver enzyme metabolic condition simulation is better; and 3. establishing indexes for evaluating the enzyme metabolic activity: metabolite ratios.

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