Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
J Transl Med. 2011 May 15;9:62. doi: 10.1186/1479-5876-9-62.
Berberine (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.
BBR 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.
In 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.
Four 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.
小檗碱（BBR）是一种对细胞能量代谢具有多种作用的药物。本研究探讨了哪个 CYP450（细胞色素 P450）同工酶执行 BBR 的 I 相转化，以及其代谢物在能量途径上的生物活性是什么。
使用 LC-MS/MS 检测 BBR 代谢物。计算机辅助对接技术以及用重组 CYP450 进行的生物测定用于鉴定负责 BBR I 相转化的 CYP450 同工酶。用实时 RT-PCR 和激酶磷酸化测定法检测 BBR 代谢物在肝细胞中的生物活性。
在大鼠实验中，使用 LC-MS/MS（液相色谱-串联质谱）在大鼠肝脏中鉴定出 BBR 的 4 种主要代谢物，小檗红碱（M1）、唐莲芬定（M2）、去亚甲二氢小檗碱（M3）和小檗碱（M4）。在无细胞转化反应中，孵育 BBR 与 rCYP450 或人肝微粒体后可检测到 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 参与 M3 的产生。肝细胞培养表明，BBR 可增强胰岛素受体（InsR）和低密度脂蛋白受体（LDLR）mRNA 的表达，并激活 AMP 激活蛋白激酶（AMPK）。BBR 的代谢物 M1-M4 仍能上调 InsR 表达，活性降低 50-70%；LDLR mRNA 仅由 M1 或 M2 增加（但不是 M3 和 M4），活性水平分别为 BBR 的 35%或 26%。同样，AMPK-α 磷酸化仅由 M1 和 M2 增强，程度小于 BBR。
在大鼠肝内 I 相转化后鉴定出 BBR 的 4 种主要代谢物（M1-M4）。无细胞反应表明，CYP2D6、CYP1A2 和 CYP3A4 似乎是将 BBR 转化为代谢物 M2 和 M3 的主要 CYP450 同工酶。BBR 的代谢物在 BBR 的靶标（InsR、LDLR 和 AMPK）上仍保持活性，但活性降低。
Zotero 插件
