【摘要】：目的基于二相代謝酶UGT1A1酶介導(dǎo)膽紅素代謝過程,測定比較不同種屬肝微粒體體系中UGT1A1酶動力學(xué)參數(shù)。方法采用高分辨質(zhì)譜對膽紅素及其葡萄糖醛酸代謝物進(jìn)行定性鑒別,采用UPLC測定法,分別以膽紅素一級、二級和總代謝產(chǎn)物生成量對膽紅素底物濃度作圖,求得動力學(xué)參數(shù)。結(jié)果實驗結(jié)果發(fā)現(xiàn),以總代謝產(chǎn)物計,KRLMKHLM≈Kr UGT1A1,VRLM≈VHLMVr UGT1A1,CLHLMCLr UGT1A1≈CLrRLM。r UGT1A1(人重組UGT1A1酶)酶體系中UGT1A1酶Km值最小,表明其與膽紅素親和能力最強,Vmax值最小,不能及時將其代謝轉(zhuǎn)化;與此相反,大鼠肝微粒體(RLM)體系中UGT1A1酶與膽紅素的親和能力最弱,但轉(zhuǎn)化效果最佳;人肝微粒體(HLM)體系中UGT1A1酶對膽紅素親和能力較強,轉(zhuǎn)化能力適中。結(jié)論通過3個體系比較發(fā)現(xiàn),人肝微粒體與r UGT1A1體系Km值接近,確證了膽紅素在肝微粒體中的葡萄糖醛酸化反應(yīng)主要由UGT1A1介導(dǎo),人肝微粒體和大鼠肝微粒體體系差別主要由酶及其環(huán)境的種屬差異導(dǎo)致,本實驗的相關(guān)數(shù)據(jù)為研究因誘導(dǎo)或抑制UGT1A1酶引起的藥物相互作用及毒性提供了實驗依據(jù)。
[Abstract]:Aim based on the bilirubin metabolism mediated by biphasic metabolizing enzyme UGT1A1 enzyme, the kinetic parameters of UGT1A1 enzyme in liver microsomal system of different species were determined and compared. Methods bilirubin and its glucuronic acid metabolites were identified qualitatively by high resolution mass spectrometry (HRMS). UPLC was used to determine the concentration of bilirubin substrate, and the kinetic parameters were obtained by mapping the concentration of bilirubin substrate with the first, second and total metabolites of bilirubin respectively. Results the results showed that the km value of UGT1A1 enzyme was the lowest in KRLMKHLM 鈮，

本文編號：2471137