紫堇药效成分木兰花碱调控AMPKα改善高脂血症的分子机制研究

本项目前期以“痰浊证”和“祛湿化浊”中医理论为指导对紫堇进行药效评价，证实紫堇总生物碱能显著改善高脂血脂和动脉粥样硬化。在此基础上，通过对紫堇系统的化学成分、体外活性筛选、体内暴露量评价和体内活性验证，发现木兰花碱是其改善高脂血症和动脉粥样硬化的主要药效成分；初步机制研究发现，木兰花碱能显著激活肝细胞AMPKα磷酸化并抑制肝脏分泌极低密度脂蛋白（VLDL），推测AMPKα可能是调控VLDL分泌的全新靶标。本项目以木兰花碱为工具，利用靶酶、细胞、基因敲除和过表达细胞与动物模型，深入揭示AMPKα/CLOCK/MTP通路调控VLDL分泌的分子机制，阐明以木兰花碱为代表的紫堇生物碱类成分通过“祛湿化浊”治疗高脂血症的作用靶点和调控通路，从脂蛋白代谢的角度探讨中医“痰浊证”和“祛湿化浊”疗法的科学内涵，对于中医药理论阐释和新型抗高脂血症药物研发都具有重要意义。

Guided by the traditional Chinese medical theories in terms of “turbid phlegm symptom” and “eliminating dampness and turbidity,” the therapeutic effects of the total corydalis alkaloid extract of Corydalis edulis Maxim. on hyperlipidemia have been disclosed in our previous study. We found that the corydalis alkaloids were able to significantly decrease the plasma lipids levels, indicating promising anti-atherosclerotic potential. Subsequently, extensive efforts, such as the chemical composition characterization, in vitro activity investigation, in vivo exposure evaluation, and in vivo activity verification were made for those corydalis alkaloids. We demonstrated that magnoflorine from corydalis is the main effective ingredient being responsible for anti-hyperlipidemia and anti-atherosclerosis. The preliminary findings have demonstrated that magnoflorine could significantly activate AMPKα phosphorylation in liver cells and inhibit the secretion of the very low-density lipoprotein (VLDL) from the liver, indicating that AMPKα should be involved in a novel pathway for regulating VLDL. In this project, we will use magnoflorine as a tool to explore the molecular mechanism and regulation of the VLDL secretion by the AMPKα/CLOCK/MTP pathway using target enzymes, gene knockout, and overexpression from cells and animal models. We also aim to clarify the target and molecular mechanism of the magnoflorine being responsible for the anti-hyperlipidemia benefits according to the “eliminating dampness and turbidity” theory. The molecular mechanism clarification of lipoprotein metabolism towards “eliminating dampness and turbidity” is envisioned to be of great importance towards the interpretation of Chinese medicine theory and the development of new anti-hyperlipidemic drugs as well.