补骨脂酚在大鼠和人肝微粒体中细胞色素P450酶和尿苷二磷酸葡萄糖醛酸转移酶代谢稳定性及性别差异

Gender differences in cytochrome P450 enzyme and UDP-glucuronosyltransferase mediated metabolic stabilities of bakuchiol in rat and human liver microsomes

目的探究补骨脂酚在大鼠和人肝微粒体中细胞色素P450酶(CYP酶)和尿苷二磷酸葡萄糖醛酸转移酶(UGT酶)的代谢稳定性及性别差异。方法补骨脂酚分别与雄、雌性SD大鼠和男、女性人肝微粒体在37℃与不同辅酶因子孵育,应用高效液相色谱(HPLC)法测定补骨脂酚的剩余浓度,采用底物消除法观察补骨脂酚的代谢稳定性。结果补骨脂酚在雄、雌性SD大鼠肝微粒体中,CYP酶介导的Ⅰ相代谢固有清除率(Clint)分别是326.6±15.4和(77.2±4.8)mL·min-1·kg-1,雄性代谢显著快于雌性(P<0.01);UGT酶介导的Ⅱ相代谢Clint分别是164.5±8.4和(419.1±24.1)mL·min-1·kg-1,雌性代谢显著快于雄性(P<0.01);CYP酶和UGT酶共同代谢的Clint分别是1063.1±27.2和(781.2±16.5)mL·min-1·kg-1,雄性代谢显著快于雌性(P<0.01)。在男、女性人肝微粒体中,CYP酶介导的Ⅰ相代谢Clint分别是24.8±2.1和(17.6±1.0)mL·min-1·kg-1,男性代谢显著快于女性(P<0.01);UGT酶介导的Ⅱ相代谢Clint分别是176.4±26.5和(165.9±8.6)mL·min-1·kg-1,代谢无显著性别差异;CYP酶和UGT酶共同代谢的Clint分别是262.5±20.9和(236.2±10.5)mL·min-1·kg-1,代谢无显著性别差异。结论补骨脂酚在SD大鼠和人肝微粒体中,均发生CYP酶介导的Ⅰ相代谢和UGT酶介导的Ⅱ相代谢反应,且代谢稳定性具有一定的种属和性别差异。

OBJECTIVE To investigate the metabolic characteristics of bakuchiol mediated by cytochrome P450 enzyme(CYP)and UDP-glucuronosyltransferase(UGT)in rat liver microsomes(RLMs)or human liver microsomes(HLMs),and to compare the metabolic gender differences.METHODS Bakuchiol was incubated at 37℃with male and female RLMs or HLMs in the presence of nicotinamide adenine dinucleotide phosphate(NADPH)or uridine 5′-diphosphoglucuronic acid(UDPGA).The residual concentrations of bakuchiol were measured in each incubation system using high performance liquid chromatography(HPLC).The metabolic stability and metabolic gender differences of bakuchiol were evaluated by the remaining percentage of bakuchiol after incubation.RESULTS When bakuchiol was metabolized by CYP in RLMs,the intrinsic clearance(Clint)value in male RLMs〔(326.6±15.4)mL·min-1·kg-1〕was significantly higher than that of female RLMs〔(77.2±4.8)mL·min-1·kg-1〕(P<0.01).When bakuchiol was metabolized by UGT in RLMs,female RLMs had a significantly higher Clintvalue〔(419.1±24.1)mL·min-1·kg-1〕than male RLMs〔(164.5±8.4)mL·min-1·kg-1〕(P<0.01).When bakuchiol was metabolized by both CYP and UGT in RLMs,male RLMs had a significantly higher Clintvalue〔(1063.1±27.2)mL·min-1·kg-1〕than female RLMs〔(781.2±16.5)mL·min-1·kg-1〕(P<0.01).When bakuchiol was metabolized by CYP in HLMs,male HLMs had a significantly higher Clintvalue〔(24.8±2.1)mL·min-1·kg-1〕than female HLMs〔(17.6±1.0)m L·min-1·kg-1〕(P<0.01).There were no significant gender differences in the metabolism of bakuchiol when it was metabolized by UGT in HLMs.The Clintvalues were 176.4±26.5 and(165.9±8.6)mL·min-1·kg-1,respectively.The metabolic parameters of bakuchiol mediated by CYP and UGT in HLMs had no significant gender differences.The Clintvalues were 262.5±20.9 and(236.2±10.5)mL·min-1·kg-1,respectively.CONCLUSION Bakuchiol can be metabolized by CYP and UGT in RLMs or HLMs,and the metabolic parameters exhibit species differences and gender differences.