该研究通过高效液相色谱法（High Performance Liquid Chromatography，HPLC）测定荷叶中荷叶碱含量，对比分析25种荷叶中荷叶碱含量差异，并利用油酸诱导建立AML-12细胞脂质蓄积模型，研究荷叶碱对脂质代谢的影响。通过测定甘油三酯（Triglyceride，TG)含量和油红O染色情况，评估细胞脂滴蓄积情况；检测细胞内AMP依赖的蛋白激酶（Adenosine 5‘-Monophosphate-Activated Protein Kinase，AMPK）通路相关蛋白表达水平，探究荷叶碱影响脂质代谢的作用机制。经HPLC检测，发现荷叶品种间荷叶碱含量差异显著，其中佛座莲的荷叶碱质量分数最高，达0.68 mg/g，而金凤的荷叶碱质量分数最低，仅为 0.22 mg/g。在脂质代谢研究中，与对照组相比，模型组在建模后脂滴数量显著增加，TG含量提升（P＜0.01），P-AMPK和脂肪甘油三酯脂肪酶（Adipose Triglyceride Lipase，ATGL）在蛋白水平上表达显著降低（P＜0.05）；在荷叶碱的干预下，AML-12细胞的脂滴蓄积程度明显降低，TG含量显著减少（P＜0.05），P-AMPK和ATGL的蛋白表达水平显著升高（P＜0.05）。由此表明：不同荷叶品种间荷叶碱含量存在明显差异，荷叶碱可能是通过调控AMPK/ATGL途径调节AML-12细胞内脂质代谢。该结果可为荷叶的开发利用提供理论支持。

Nuciferine content in 25 varieties of lotus leaf was determined using high-performance liquid chromatography (HPLC). A lipid accumulation model using AML-12 cells was established with oleic acid induction to study the effects of nuciferine on lipid metabolism. The accumulation of lipid droplets in the cells was evaluated by measuring triglyceride (TG) content and by oil red O staining, and the expression of proteins regulated by the adenosine 5'-monophosphate-activated protein kinase (AMPK) pathway was assessed to explore the mechanism by which nuciferine affects lipid metabolism. HPLC analysis revealed significant differences in nuciferine content among different lotus leaf varieties. Among these variants, the mass fraction from “Fozuolian” was highest (0.68 mg/g), while that of “Jinfeng” was lowest (0.22 mg/g). In terms of lipid metabolism, compared with the control group, the number of lipid droplets in the model group increased significantly with increasing TG level, while P-AMPK and adipose triglyceride lipase (ATGL) protein expression decreased significantly (P<0.05). With administration of nuciferine, lipid droplet and TG content accumulation in AML-12 cells significantly decreased (P<0.05), while expression of P-AMPK and ATGL proteins significantly increased (P<0.05). This suggests that there are significant differences in nuciferine content among lotus leaves of different varieties, and nuciferine may regulate lipid metabolism in AML-12 cells by regulating the AMPK/ATGL pathway. These results provide technical support for the development and utilization of lotus leaves.

浙江省重点研发计划项目（2021C02018）；浙江中医药大学校级科研项目（2020ZG08；2020ZR07）