目的 基于细胞药动学探究斑蝥素与黄芩苷配伍在亚细胞水平的分布规律，阐明其抗肝癌的增效机制。方法 以人肝癌HepG2细胞为模型，采用超高效液相色谱-串联质谱（ultra-high performance liquid chromatography-tandem mass spectrometry，UPLC-MS/MS）定量分析单药（斑蝥素6 μg/mL、黄芩苷30 μg/mL）及配伍组（斑蝥素6 μg/mL＋黄芩苷30 μg/mL）给药后12 h内，整体细胞及细胞核、线粒体、内质网和溶酶体各细胞器中药物浓度的动态变化，并应用Phoenix WinNonlin软件非房室模型计算药动学参数。结果 在整体细胞水平，配伍使斑蝥素的胞内药时曲线下面积（area under the curve，AUC_0～t）增加48.9%，清除率降低（P＜0.05），但未显著影响黄芩苷的药动学行为。在亚细胞层面，配伍使斑蝥素与黄芩苷在细胞核、溶酶体、线粒体、内质网内的AUC_0～t分别增加了93.5%、46.4%、38.3%、52.3%和68.4%、40.0%、41.0%、46.7%（P＜0.05、0.01）。此外，配伍后2种药物在线粒体内达峰时间（t_max）提前，且斑蝥素在内质网中的平均驻留时间（mean residence time，MRT_0～t）显著延长（P＜0.01），表明两者配伍实现了时空协同的药物递送。结论 斑蝥素/黄芩苷配伍可通过协同优化药物在细胞核、线粒体、内质网及溶酶体等关键亚细胞结构的分布，进而可能通过诱导DNA损伤、加速线粒体介导细胞凋亡及促进内质网应激等途径，增强抗肝癌效果，为基于细胞器靶向的中药配伍设计提供了理论依据。

Objective To investigate the subcellular distribution patterns of the combination of cantharidin (CTD) and baicalin (BA) based on cell pharmacokinetics and elucidate the synergistic mechanism against hepatocellular carcinoma. Methods Using human hepatoma HepG2 cells as the model, the dynamic concentrations of the drugs in whole cells and organelles (nucleus, mitochondria, endoplasmic reticulum, and lysosomes) over 12 h after administration of single drugs (CTD 6 μg/mL, BA 30 μg/mL) and their combination (CTD 6 μg/mL + BA 30 μg/mL) were quantitatively analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Pharmacokinetic parameters were calculated using a non-compartmental model with Phoenix WinNonlin software. Results At the whole-cell level, the combination increased the intracellular area under the curve (AUC_0—t) of CTD by 48.9% and decreased its clearance (P < 0.05), but did not significantly affect the pharmacokinetics of BA. At the subcellular level, the combination significantly increased the AUC_0—t of CTD and BA in nucleus, lysosome, mitochondria and endoplasmic reticulum by 93.5%, 46.4%, 38.3%, 52.3%, and 68.4%, 40.0%, 41.0%, and 46.7%, respectively (P < 0.05, 0.01). Furthermore, the time to reach peak concentration (t_max) for both drugs in mitochondria was earlier in the combination group, and the mean residence time (MRT_0—t) of CTD in the endoplasmic reticulum was significantly prolonged (P < 0.01), indicating a spatiotemporally synergistic drug delivery optimization. Conclusions The combination of CTD and BA could synergistically optimize the distribution of drugs in key subcellular structures such as nucleus, mitochondria, endoplasmic reticulum and lysosomes, and may enhance the anti-hepatocellular carcinoma effect by inducing DNA damage, accelerating mitochondria-mediated apoptosis and promoting endoplasmic reticulum stress, which provides a theoretical basis for the design of traditional Chinese medicine combinations based on organelle targeting.

R285.61

湖南省卫生健康委员会项目(202213014042);湖南省教育厅优秀青年项目(22B0375);湖南省自然科学基金项目(2022JJ80023);国家中医药管理局全国老药工传承工作室建设项目(国中医药人教函[2024]255号)