吴茱萸生物碱经CAR调节糖脂代谢的机制及其PD/PK研究

Currently, lipid metabolism disorders secondary to diabetes have no good therapeutic targets. It was found that constitutive androstane receptor (CAR) has a role in regulating glucose and lipid metabolism, but the mechanism is not clear, it is likely to become a new effective therapeutic targets for such diseases, is the focus of current research. Our previous studies have found evodia rutaecarpa alkaloids significantly lowering blood pressure, lowering blood sugar and increase the role of insulin sensitivity, and CAR may play an important role in which the results of in vitro and in vivo experiments speculated. In addition, evodia rutaecarpa alkaloids can induce some metabolic enzymes, precisely in itself may be metabolized by these enzymes. Therefore, the subject is to be adopted gene knock-out mice, high-fat diabetic mice, primary hepatocytes cells in vitro and in vivo models using co-immunoprecipitation and gel mobility shift assay technology and some other technologies to explore the role of CAR in the evodia rutaecarpa alkaloids regulating glucose and lipid metabolism and elucidating the mechanisms. Moreover, ascertain the pharmacokinetics difference between the disease states and the normal state, as well as during long-term repeated administration of evodia rutaecarpa alkaloids the enzyme induction characteristics and its impact on the pharmacokinetic behavior and pharmacodynamics, clarify PD/PK correlation and the effect of CAR in this process, so as to provide a theoretical basis for the in-depth development of the evodia rutaecarpa alkaloids.

脂代谢紊乱继发糖尿病目前尚没有良好的治疗靶点。研究发现组成型雄甾烷受体（CAR）具有调节糖脂代谢的作用，但机制尚不明确，其很可能成为该类疾病新的有效治疗靶点，是目前研究的热点。我们前期的研究发现吴茱萸生物碱具有显著的降血脂、降血糖和增加胰岛素敏感性的作用，而且根据体内外实验结果推测CAR在其中可能发挥了重要的作用。此外，吴茱萸生物碱对一些药物代谢酶具有诱导作用，而其自身恰恰可能通过这些酶进行代谢而影响其药理作用。因此，本课题拟采用基因敲除小鼠，高脂糖尿病小鼠，原代肝细胞等体内外模型、应用免疫共沉淀和凝胶迁移率变动分析等技术探究CAR在吴茱萸生物碱调节糖脂代谢中的作用并深入阐明相关机制；进而弄清疾病状态和正常状态下其药动学行为的差异，以及长期重复给药过程中药酶诱导的特性及其对药动学行为和药效的影响，阐明PD/PK的相关性及CAR在其中的作用，从而为吴茱萸生物碱的深入开发提供理论基础。

脂代谢紊乱继发糖尿病还没有良好的治疗靶点。研究发现组成型雄甾烷受体（CAR）具有调节糖脂代谢的作用，但机制尚不明确，其很可能成为该类疾病新的有效治疗靶点，是目前研究的热点。我们前期的研究发现吴茱萸生物碱具有显著的降血脂、降血糖和增加胰岛素敏感性的作用，而且通过体内外实验结果推测CAR在其中可能发挥了重要的作用。因此，本课题拟采用基因敲除小鼠，高脂糖尿病小鼠，原代肝细胞等体内外模型探究CAR在吴茱萸生物碱调节糖脂代谢中的作用并深入阐明相关机制；进而弄清疾病状态和正常状态下药动学行为的差异，以及长期重复给药过程中药酶诱导的特性及其对药动学行为和药效的影响，阐明PD/PK的相关性，从而为吴茱萸生物碱的深入开发提供理论基础。我们发现吴茱萸次碱和吴茱萸碱在HepG2高脂细胞模型上都表现了明显的降糖降脂作用。这两个化合物都能显著激活CAR，而CAR的抑制剂能明显逆转这种作用。吴茱萸次碱和吴茱萸碱的抑制糖异生作用是因为这两个化合物激活CAR后，抑制了FoxO1和HNF4α与糖异生相关基因PEPCK和G6Pase基因启动子区域的结合而导致的。体内实验发现，吴茱萸次碱给药小鼠后增加了其糖耐受性，而这呈现CAR依赖性。我们的结果表明吴茱萸碱和次碱可能是治疗二型糖尿病的潜在新药。该研究成果发表在BBA Gene Regulatory Mechanisms上。