基于细胞色素P4502B6酶的补骨脂素对环泊酚代谢的影响

Effects of psoralen on the metabolism of ciprofol based on cytochrome P450 2B6 enzyme

目的通过体外大鼠肝微粒体孵育实验考察环泊酚的酶促反应动力学,并进一步探究补骨脂素对其代谢的影响。方法色谱柱:Diamonsil C18色谱柱(250.0 mm×4.6 mm,5μm),柱温:30℃,流动相:乙腈-水=73∶27,检测波长:272 nm,流速:1.0 mL·min^(-1),进样量:20μL,内标为丙泊酚。大鼠肝微粒体与系列浓度的环泊酚注射液共同孵育,用Graph Pad Prism 8软件计算环泊酚的米氏常数(Km)、最大反应速度(Vmax)和固有清除率(CLint);孵育体系中加入系列浓度的补骨脂素,用HPLC法测定大鼠肝微粒体中的环泊酚减少量,并计算半数抑制浓度(IC50),考察补骨脂素对环泊酚代谢的影响。结果环泊酚在大鼠肝微粒体孵育体系中的Km为129.8μmol·L^(-1),Vmax为98.42 nmol·min^(-1)·mg protein^(-1),CLint为0.76 mL·min^(-1)·mg^(-1)。补骨脂素对环泊酚的代谢呈剂量依赖性抑制作用,IC50值为63.31μmol·L^(-1)。结论补骨脂素可降低环泊酚在大鼠肝微粒体内的代谢,延长麻醉苏醒时间。

Objective To investigate the enzymatic reaction kinetics of ciprofol by incubation experiment of rat liver microsomes in vitro and further explore the effects of psoralen on its metabolism.Methods HPLC conditions:A Diamonsil C18 column(250.0 mm x 4.6 mm,5μm)was used and the column temperature was set at 30 C.The mobile phase consisted of acetonitrile-water(73:27)at a flow rate of 1.0 mL·min^(-1).The detection wavelength was 272 nm,and the injection volume was 20μL.Propofol was used as internal standard.An in vitro incubation system of rat liver microsomes was established and incubated with serial concentrations of ciprofol injection.The michaelis constant(K.),maximum reaction rate(V_(max))and intrinsic clearance(CLm)of ciprofol were calculated with Graph Pad Prism 8 software.Next,a series of concentrations of psoralen was co-incubated with ciprofol injection,the decrease of ciprofol in rat liver microsomes was determined by HPLC,and the median inhibitory concentration(IC)was calculated to investigate the effect of psoralen on the metabolism of ciprofol.Results In the rat liver microsome incubation system,the Km,V,and CLm of eiprofol were 129.8 umol·L^(-1),98.42 nmol·min^(-1)·mg protein^(-1)and 0.76 mL·min^(-1)mg,respectively.Psoralen could inhibit the metabolism of ciprofol in rat liver microsomes in a dose-dependent manner with ICso value of 63.31μmol·L^(-1).Conclusion Psoralen can inhibit the metabolism of ciprofol in rats,prolong the recovery time of anesthesia.