目的 基于体外耐药细胞模型，研究白屈菜-元胡药对（Chelidonii Herba-Corydalis Rhizoma drug pair，CMCR）逆转乳腺癌阿霉素（adriamycin，ADR）耐药药效物质。方法 运用超高效液相色谱-四极杆飞行时间质谱联用技术（UPLC-Q-TOF-MS）对CMCR不同配伍比例（1∶1、2∶1）95%、80%、70%、60%、50%乙醇提取部位的化学成分进行表征，确定共有成分；以人乳腺癌阿霉素耐药细胞（MCF-7/ADR）为研究对象，采用CCK-8法检测ADR联合CMCR各提取部位对MCF-7/ADR细胞增殖的影响，计算耐药倍数；利用灰色关联度、Pearson相关性分析和偏最小二乘回归分析建立表征图谱中共有成分峰面积与逆转耐药倍数的相关性，筛选出CMCR最佳逆转耐药活性部位及相关药效物质。采用荧光显微镜观察白屈菜碱联合延胡索乙素作用细胞后罗丹明123累积量的影响；Western blotting和qRT-PCR法检测P-糖蛋白（P-glycoprotein，P-gp）、乳腺癌耐药蛋白（breast cancer resistant protein，BCRP）、肺耐药蛋白（lung resistance protein，LRP）蛋白和基因表达。结果 CMCR 10个提取部位分析得到30个共有成分。CMCR（1∶1）95%乙醇提取部位对ADR的逆转耐药活性最强，其逆转耐药倍数为11.08。根据相关性分析结果初步确定白屈菜碱、延胡索乙素、小檗碱、二氢血根碱和白屈菜红碱为CMCR逆转乳腺癌ADR耐药的潜在药效物质。选择药效相关性最高的白屈菜碱和延胡索乙素联合进行实验验证，发现白屈菜碱-延胡索乙素能够增加细胞内罗丹明123累积，显著下调MCF-7/ADR细胞P-gp、BCRP和LRP蛋白和基因表达（P＜0.01、0.001）。结论 阐明了CMCR逆转乳腺癌阿霉素耐药的药效物质，为其临床应用及后续抗肿瘤辅助用药开发奠定了基础。

Objective To study the pharmacological substances of Baiqucai (Chelidonii Herba)-Yuanhu (Corydalis Rhizoma) drug pair (CMCR) in reversing breast cancer resistance to adriamycin (ADR) based on the drug-resistant cell model in vitro. Methods UPLC-Q-TOF-MS technology was used to characterize the chemical components of 95%, 80%, 70%, 60% and 50% ethanol-extracted parts of CMCR (compatibility proportion 1∶1, 2∶1) and determine the common components; CCK-8 was applied to detect the effect of ADR combined with CMCR on the proliferation of MCF-7/ADR cells to calculate the drug resistance multiples; Grey correlation analysis method, Pearson bivariate correlation analysis and partial least squares regression analysis were used to establish the correlation between the peak area of common peak and the multiple of reversal resistance in the characterization map to select the best active site of reverse resistance and related medicinal substances of CMCR. Fluorescence microscope was employed to investigate the influence of the combination of chelidonine (CHE) and tetrahydropalmatine (THP) on the intracellular accumulation of rhodamine 123 (Rh123). Western blotting and qRT-PCR were used to detect P-glycoprotein (P-gp), breast cancer resistance protein (BCRP), lung resistance protein (LRP) protein and gene expressions. Results A total of 30 common components were analyzed from 10 extracted parts of CMCR. The CMCR (1∶1) 95% ethanol extracted part exhibited the most potent reversing ADR resistance activity, with a reversal drug resistance multiple of 11.08. According to the initial findings of the correlation analysis results, chelidonine, tetrahydropalmatine, berberine, dihydrosanguinarine and chelerythrine were preliminarily determined as pharmacodynamic substances for CMCR to reverse ADR resistance in BC. Since CHE and THP had the highest correlation in pharmacodynamic efficacy, they were paired to conduct a verification experiment, which revealed that CHE-THP could increase the intracellular accumulation of Rh123 and significantly down-regulate P-gp, BCRP, LRP protein and gene expressions in MCF-7/ADR cells (P < 0.01, 0.001). Conclusion This study elucidated the effective substances of CMCR reversing ADR resistance in BC and laid the foundation for its clinical application and subsequent development of anti-tumor adjuvant drugs.

黑龙江省自然科学基金资助项目（LH2022H001）