目的 研究包载三苯基膦-阿霉素（TPP-DOX，TD）和槲皮素（Que）的纳米混合胶束的体外抗肿瘤耐药与线粒体靶向作用。方法 以还原敏感性聚合物材料聚乙二醇-脱氧胆酸-二硫键-聚天冬氨酸苄酯（mPEG-DCA-SS-PBLA，PDSP）和非还原敏感性聚合物材料聚乙二醇-脱氧胆酸-碳碳键-聚天冬氨酸苄酯（PDCP）为载体，通过溶剂挥发法分别包载TD和Que，制备还原敏感性纳米胶束PDSP@TD、PDSP@Que和非还原敏感性纳米胶束PDCP@TD、PDCP@Que。通过MTT法考察游离TD、DOX对人乳腺癌DOX耐药细胞株MCF-7/ADR的耐药性，考察TD与DOX、PDSP@TD与PDSP@Que作用于MCF-7/ADR细胞联合使用的最佳协同比，考察TD、Que、PDCP@TD、PDCP@Que、PDSP@TD、PDSP@Que、PDCP@TD＋ PDCP@Que、PDSP@TD＋ PDSP@Que对正常肝细胞HL-7702和MCF-7/ADR的毒性；采用HPLC法对MCF-7/ADR细胞摄取DOX、TD、Que、TD＋ Que、PDSP@TD、PDSP@Que、PDCP@TD、PDCP@Que、PDSP@TD＋PDSP@Que、PDCP@TD＋PDCP@Que结果进行定量测定；通过对MCF-7/ADR细胞核及线粒体染色、细胞内ROS及线粒体跨膜电位检测来验证胶束的线粒体靶向性。结果 TD、Que、DOX对MCF-7/ADR细胞的耐药指数（RI）分别为126.76、2.54、1.65；游离药物TPP-DOX和Que以及纳米胶束PDSP@TD和PDSP@Que在质量比分别为1∶ 2和1∶ 1时均具有较强的协同作用； PDSP@TD＋ PDSP@Que相比于游离TD和Que对HL-7702细胞的毒性显著降低（P＜ 0.05、0.01、0.001），对MCF-7/ADR细胞毒性较TD及PDCP@TD＋ PDCP@Que组显著增强（P＜ 0.05、0.01）； MCF-7/ADR细胞对PDSP@TD＋PDSP@Que的摄取率最高； DOX给药后主要聚集在细胞核中，PDSP@TD＋PDSP@Que给药后DOX大多数都聚集在线粒体内，细胞内ROS含量明显增加，较DOX、TD、PDSP@TD对线粒体膜电位的破坏作用更强（P＜0.05、0.01、0.001）。结论 PDSP@TD＋PDSP@Que相较于游离药物具有较好的生物安全性及抗肿瘤活性，能较好的逆转MCF-7/ADR细胞对DOX的耐药，且具有较好的线粒体靶向性。

Objective To study the in vitro anti-tumor resistance and mitochondrial targeting effects of reduction sensitive nano mixed micelles loaded with triphenylphosphine-doxorubicin (TPP-DOX, TD) and quercetin (Que). Methods The redox-sensitive polymer material poly (ethyleneglycol)-deoxycholic acid-disulfide-poly (aspartic acid benzyl ester), PDSP, and the non-redox-sensitive polymer material poly (ethylene glycol)-deoxycholic acid-carbon-carbon bond-poly(aspartic acid benzyl ester), PDCP, were used as the carrier. TD and Que were loaded onto the polymers by solvent evaporation, respectively, to prepare redox-sensitive nanovesicles PDSP@TD and PDSP@Que, and non-redox-sensitive nanovesicles PDCP@TD and PDCP@Que. The resistance of human breast cancer DOXresistant cell line MCF-7/ADR to free TD and DOX was investigated by MTT assay. The optimal synergistic ratio of TD and DOX, PDSP@TD and PDSP@Que in combination with MCF-7/ADR cells was examined. The toxicity of TD, Que, PDCP@TD, PDCP@Que, PDSP@TD, PDSP@Que, PDCP@TD + PDCP@Que, and PDSP@TD + PDSP@Que on normal liver cells HL-7702 and MCF-7/ADR was evaluated. The uptake of DOX, TD, Que, TD+Que, PDSP@TD, PDSP@Que, PDCP@TD, PDCP@Que, PDSP@TD + PDSP@Que, and PDCP@TD + PDCP@Que by MCF-7/ADR cells was quantitatively determined by HPLC. The mitochondrial targeting of the micelles was verified by nuclear and mitochondrial staining of MCF-7/ADR cells, intracellular ROS detection, and mitochondrial membrane potential measurement. Results The resistance indices (RI) of TD, Que, and DOX to MCF- 7/ADR cells were 126.76, 2.54, and 1.65, respectively. Free drugs TPP-DOX and Que, as well as nanomicelles PDSP@TD and PDSP@Que, showed strong synergistic effects at mass ratios of 1:2 and 1:1, respectively. The toxicity of PDSP@TD + PDSP@Que to HL-7702 cells was significantly lower than that of free TD and Que (P < 0.05, 0.01, 0.001), and the toxicity to MCF-7/ADR cells was significantly higher than that of TD and PDCP@TD + PDCP@Que (P < 0.05, 0.01). The uptake rate of PDSP@TD + PDSP@Que by MCF-7/ADR cells was the highest. After DOX administration, it mainly accumulated in the cell nucleus, while after PDSP@TD + PDSP@Que administration, most of the DOX accumulated in the mitochondria, and the intracellular ROS content significantly increased. The damage to the mitochondrial membrane potential was stronger than that of DOX, TD, and PDSP@TD (P < 0.05, 0.01, 0.001). Conclusion PDSP@TD + PDSP@Que has better biocompatibility and antitumor activity than free drugs, can better reverse the resistance of MCF-7/ADR cells to DOX, and has better mitochondrial targeting.

R285.5

河南省科技攻关项目（232102311178）;河南省高等学校重点科研项目（25B350003）;河南应用技术职业学院名师工作室项目（2022-03）;河南应用技术职业学院教师创新团队项目（2022-02）