Bioactivities of berberine metabolites after transformation through CYP450 isoenzymes.
Bioactivities of berberine metabolites after transformation through CYP450 isoenzymes.
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CYP450同工酶转化小檗碱代谢物的生物活性
DOI：
10.1186/1479-5876-9-62
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发表时间：
2011-05-15
影响因子：
7.4
通讯作者：
Jiang JD
中科院分区：
文献类型：
--
作者：
Li Y;Ren G;Wang YX;Kong WJ;Yang P;Wang YM;Li YH;Yi H;Li ZR;Song DQ;Jiang JD
研究背景小檗碱（BBR）是一种对细胞能量代谢具有多种作用的药物。本研究探讨了答案的问题，CYP450（细胞色素P450）同工酶执行的第一阶段转换BBR，以及什么是其代谢产物的生物活性的能量pathways. MethodsBBR代谢产物检测使用LC-MS/MS。计算机辅助对接技术以及生物测定与重组CYP450的，以确定CYP450同工酶负责BBR第一阶段转换。结果采用液相色谱-串联质谱法（LC-MS/MS），在大鼠肝组织中鉴定出小檗红碱（M1）、噻苯芬定（M2）、去甲小檗碱（M3）和药根碱（M4）4种主要代谢产物。在无细胞转化反应中，BBR与rCYP 450或人肝微粒体孵育后可检测到M2和M3;然而，M1和M4低于检测水平。CYP2D6和CYP1A2在将BBR转化为M2中起主要作用; CYP2D6、CYP1A2和CYP3A4用于M3的产生。肝细胞培养结果显示，BBR能增强胰岛素受体（InsR）和低密度脂蛋白受体（LDLR）mRNA的表达，并能激活AMP活化蛋白激酶（AMPK）。BBR的代谢产物M1-M4在上调InsR表达方面仍具有活性，效力降低50 - 70%;仅M1或M2（而非M3和M4）增加LDLR mRNA，活性水平分别为BBR的35%或26%。同样地，AMPK-α磷酸化仅被M1和M2所增强，其程度小于BBR。结论在大鼠肝脏中，经I相转化后，鉴定出4种主要的BBR代谢产物（M1-M4）。无细胞反应显示，CYP2D6、CYP1A2和CYP3A4似乎是将BBR转化为其代谢产物M2和M3的主要CYP450同工酶。BBR的代谢产物对BBR的靶点（InsR、LDLR和AMPK）仍有活性，但效力降低。
BackgroundBerberine (BBR) is a drug with multiple effects on cellular energy metabolism. The present study explored answers to the question of which CYP450 (Cytochrome P450) isoenzymes execute the phase-I transformation for BBR, and what are the bioactivities of its metabolites on energy pathways.MethodsBBR metabolites were detected using LC-MS/MS. Computer-assistant docking technology as well as bioassays with recombinant CYP450s were employed to identify CYP450 isoenzymes responsible for BBR phase-I transformation. Bioactivities of BBR metabolites in liver cells were examined with real time RT-PCR and kinase phosphorylation assay.ResultsIn rat experiments, 4 major metabolites of BBR, berberrubine (M1), thalifendine (M2), demethyleneberberine (M3) and jatrorrhizine (M4) were identified in rat's livers using LC-MS/MS (liquid chromatography-tandem mass spectrometry). In the cell-free transformation reactions, M2 and M3 were detectable after incubating BBR with rCYP450s or human liver microsomes; however, M1 and M4 were below detective level. CYP2D6 and CYP1A2 played a major role in transforming BBR into M2; CYP2D6, CYP1A2 and CYP3A4 were for M3 production. The hepatocyte culture showed that BBR was active in enhancing the expression of insulin receptor (InsR) and low-density-lipoprotein receptor (LDLR) mRNA, as well as in activating AMP-activated protein kinase (AMPK). BBR's metabolites, M1-M4, remained to be active in up-regulating InsR expression with a potency reduced by 50-70%; LDLR mRNA was increased only by M1 or M2 (but not M3 and M4) with an activity level 35% or 26% of that of BBR, respectively. Similarly, AMPK-α phosphorylation was enhanced by M1 and M2 only, with a degree less than that of BBR.ConclusionsFour major BBR metabolites (M1-M4) were identified after phase-I transformation in rat liver. Cell-free reactions showed that CYP2D6, CYP1A2 and CYP3A4 seemed to be the dominant CYP450 isoenzymes transforming BBR into its metabolites M2 and M3. BBR's metabolites remained to be active on BBR's targets (InsR, LDLR, and AMPK) but with reduced potency.
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影响因子：
37.8
作者：
Bellosta, S;Paoletti, R;Corsini, A
通讯作者：
Corsini, A
影响因子：
9.8
作者：
Kong, Wei-Jia;Zhang, Hao;Jiang, Jian-Dong
通讯作者：
Jiang, Jian-Dong
影响因子：
9.8
作者：
Kong, Wei-Jia;Wei, Jin;Jiang, Jian-Dong
通讯作者：
Jiang, Jian-Dong
影响因子：
4.8
作者：
Rowland, P;Blaney, FE;Bridges, AM
通讯作者：
Bridges, AM
影响因子：
3.9
作者：
Bjornsson, TD;Callaghan, JT;Wrighton, SA
通讯作者：
Wrighton, SA