补骨脂酚的体外肝微粒体代谢及代谢减毒作用的种属比较

补骨脂酚的体外肝微粒体代谢及代谢减毒作用的种属比较焦士勇;艾常虹;李艾芳;李桦;王旗

【摘要】目的研究补骨脂酚在人、比格犬和大鼠肝微粒体的体外代谢动力学及种属差异,评价不同种属肝微粒体对补骨脂酚人肾近曲小管上皮细胞(HK-2)的减毒作用.方法应用MTT法检测HK-2细胞的存活率来评价补骨脂酚对HK-2细胞的毒性作用以及不同种属肝微粒体对其毒性的影响.应用HPLC法分析补骨脂酚在3个种属肝微粒体孵育液中的剩余浓度,研究其在肝微粒体中的代谢稳定性和代谢动力学.结果在3个种属肝微粒体分别作用下,补骨脂酚的HK-2细胞毒性明显降低.补骨脂酚在人肝微粒体中的代谢最慢,在大鼠肝微粒体中的代谢最快,人、犬和大鼠的代谢动力学参数Km、Vmax、T12)和CLint分别为:(81.66±3.41),(89.35±4.32)和(31.89±2.60)μmol·L-1;(0.47±0.01),(0.57±0.01)和(1.88±0.09) μmol·L-1·min-1;(44.14±1.13),(53.31±0.29)和(6.79±0.39)min;(39.38±1.04),(65.16±0.35)和(365.92±20.01)ml·min-1·kg-1.结论肝微粒体降低补骨脂酚的HK-2细胞毒性,这一减毒作用与补骨脂酚经肝微粒体酶的代谢相关.补骨脂酚的体外肝代谢动力学性质存在着一定的种属差异;犬肝微粒体对高浓度补骨脂酚所致的HK-2细胞毒性的降低作用要弱于人和大鼠肝微粒体.%Aim To investigate the inter-species differences of bakuchiol metabolism in human, beagle dog and rat liver microsomes by comparing enzyme kinetic parameters, and to evaluate metabolic detoxification of bakuchiol by liver microsomes from three species.Methods The cytotoxicity of bakuchiol was investigated using human kidney-2( HK-2 ),in presence or absence of liver mirosomes. The cell viabilities were examined by MTT assay. The residual concentrations of bakuchiol in microsomal incubates were determined by a HPLC method to

investigate its metabolic stability and enzyme kinetics. Results The cytotoxicity of bakuchiol was significantly attenuated in the presence of the liver microspores of all three species. The metabolic elimination of bakuchiol by human liver microsomes was the slowest among the three species,while it was rapidest in rat liver. The Km, Vmax, T1/2 and CLint of bakuchiol obtained from human, dog and rat liver microsomes were

( 81.66 ± 3.41 ),( 89.35 ± 4. 32 ) and ( 31.89 ±2.60 ) μmol · L-1 ,( 0.47 ±0. 014 ),( 0.57±0. 011 ) and ( 1.88 ±0. 087 ) μmol · L-1 · min-1,( 44. 14 ±

1.13 ),( 53.31 ±0.29 ) and ( 6.79 ±0. 39 )min,( 39.38 ± 1.04 ), ( 65.16 ± 0.35 ) and ( 365.92 ± 20.01 ) ml · min- 1 . kg-1, respectively. Conclusions The cytotoxicity of bakuchiol on HK-2 cells was attenuated by the liver microsomes from each of the three species,respectively. The detoxification was associated with the biotransformation of bakuchiol by enzymes in liver microsomes. The inter-species differences were observed in hepatic metabolic characteristics of bakuchiol. A relative weaker effect of beagle dog liver microsomes on HK-2 cytotoxicity of bakuchiol at the high concentration level was also noted in comparison to that of human and rat.【期刊名称】《中国药理学通报》

【年(卷),期】2011(027)002

【总页数】5页(P216-220)

【关键词】补骨脂酚;HK-2细胞;细胞毒性;肝微粒体;代谢;种属差异;HPLC

【作者】焦士勇;艾常虹;李艾芳;李桦;王旗

【作者单位】北京大学公共卫生学院毒理学系,北京,100191;军事医学科学院毒物

药物研究所,北京,100850;北京大学公共卫生学院毒理学系,北京,100191;军事医学

科学院毒物药物研究所,北京,100850;北京大学公共卫生学院毒理学系,北

京,100191

【正文语种】中文

【中图分类】R-332;R282.71;R322.47;R969.1

中药补骨脂为豆科植物补骨脂Psoralea corylifolia L.的干燥成熟果实，具有温肾

助阳，纳气平喘，温脾止泻的功效［1］。补骨脂酚(bakuchiol)是补骨脂的主要成分之一，在补骨脂药材中的含量为1.12%～5.14%［2］，是一种单萜类化合物，常温下为浅黄色油状液体，化学结构如Fig 1所示。补骨脂酚具有抗炎退热［3］、抗菌［4］、抗病毒［5］、抗癌［6］、保肝［7］和雌激素样作用［8］等药理活性。

Fig 1 Chemical structure of bakuchiol

张玉顺等［9］给昆明种小鼠灌胃约9.4 g·kg-1的生药后发现补骨脂酚有一定的肾毒性作用。江芳等［10］在体外也观察到20 μmol·L-1以上浓度的补骨脂酚对人

肾近曲小管上皮细胞(human kedney-2，HK-2)有明显的肾毒性作用，加入大鼠

肝匀浆S9代谢活化后，补骨脂酚的细胞毒性作用明显降低，提示肝脏药物代谢酶对补骨脂酚有一定的减毒作用，补骨脂酚经代谢可能转化为无毒或低毒的物质。补骨脂酚的代谢研究目前仅见SD大鼠口服补骨脂酚的药代动力学［11］，补骨脂

酚的肝脏代谢产物，代谢的种属差异以及代谢与毒性的关系尚未见报道。为了更好地理解补骨脂酚的肝代谢与肾毒性之间的关系及实验动物和人体之间可能存在的差异，本文用大鼠、比格犬和人肝微粒体研究了3个种属微粒体对补骨脂酚HK-2