大鼠肝细胞色素P450　2E1酶活性的气相色谱法测定

A Gas Chromatographic Method for the Measurement of the Enzyme Activity of Cytochrome P450 2E1

本文报告了用气相色谱法测定大鼠肝细胞色素Ｐ４５０２Ｅ１的酶活性。在一定的实验条件下，使丁醇在微粒体中被氧化为丁醛，用顶空气相色谱法测定产生的丁醛量，并测定肝微粒体中的蛋白质及细胞色素Ｐ４５０总量，可算出细胞色素Ｐ４５０催化丁醇氧化的速率，实验表明，乙醇诱导的细胞色素Ｐ４５０２Ｅ１明显增大了丁醇氧化的速率，而其他典型诱导剂诱导的细胞色素Ｐ４５０异构酶不增大丁醇氧化速率，所以，细胞色素Ｐ４５０催化丁醇氧化的速率可作为判断细胞色素Ｐ４５０２Ｅ１酶活性的指标。

A gas chromatographic method is presented for the measurement of the enziyme activity of cytochrome P450 2E1 in rat liver.The butanol was transformed into butyraldehyde in rat liver microsomes under the given experimental condition.Then the butyraldehyde was determined by headspace gas chromatography.The protein contents and total cytochrome P450 in liver microsomes were also determined,and then the rate of butanol oxidation catalyzed by cytochrome P450 could be calculated.The result of our study showed that cytochrome P450 2E1 induced by ethanol remarkably enhanced the rate of butanol oxidation,while the cytochrome P450 isozymes induced by other inducers did not.The rate butanol oxidation catalyzed by cytochrome P450 can be used an index in the assessment of the enzyme activity of cytochrome P450 2E1.