目的 研究包載三苯基膦-阿霉素（TPP-DOX，TD）和槲皮素（Que）的納米混合膠束的體外抗腫瘤耐藥與線粒體靶向作用。方法 以還原敏感性聚合物材料聚乙二醇-脫氧膽酸-二硫鍵-聚天冬氨酸芐酯（mPEG-DCA-SS-PBLA，PDSP）和非還原敏感性聚合物材料聚乙二醇-脫氧膽酸-碳碳鍵-聚天冬氨酸芐酯（PDCP）為載體，通過溶劑揮發(fā)法分別包載TD和Que，制備還原敏感性納米膠束PDSP@TD、PDSP@Que和非還原敏感性納米膠束PDCP@TD、PDCP@Que。通過MTT法考察游離TD、DOX對(duì)人乳腺癌DOX耐藥細(xì)胞株MCF-7/ADR的耐藥性，考察TD與DOX、PDSP@TD與PDSP@Que作用于MCF-7/ADR細(xì)胞聯(lián)合使用的最佳協(xié)同比，考察TD、Que、PDCP@TD、PDCP@Que、PDSP@TD、PDSP@Que、PDCP@TD＋ PDCP@Que、PDSP@TD＋ PDSP@Que對(duì)正常肝細(xì)胞HL-7702和MCF-7/ADR的毒性；采用HPLC法對(duì)MCF-7/ADR細(xì)胞攝取DOX、TD、Que、TD＋ Que、PDSP@TD、PDSP@Que、PDCP@TD、PDCP@Que、PDSP@TD＋PDSP@Que、PDCP@TD＋PDCP@Que結(jié)果進(jìn)行定量測(cè)定；通過對(duì)MCF-7/ADR細(xì)胞核及線粒體染色、細(xì)胞內(nèi)ROS及線粒體跨膜電位檢測(cè)來驗(yàn)證膠束的線粒體靶向性。結(jié)果 TD、Que、DOX對(duì)MCF-7/ADR細(xì)胞的耐藥指數(shù)（RI）分別為126.76、2.54、1.65；游離藥物TPP-DOX和Que以及納米膠束PDSP@TD和PDSP@Que在質(zhì)量比分別為1∶ 2和1∶ 1時(shí)均具有較強(qiáng)的協(xié)同作用； PDSP@TD＋ PDSP@Que相比于游離TD和Que對(duì)HL-7702細(xì)胞的毒性顯著降低（P＜ 0.05、0.01、0.001），對(duì)MCF-7/ADR細(xì)胞毒性較TD及PDCP@TD＋ PDCP@Que組顯著增強(qiáng)（P＜ 0.05、0.01）； MCF-7/ADR細(xì)胞對(duì)PDSP@TD＋PDSP@Que的攝取率最高； DOX給藥后主要聚集在細(xì)胞核中，PDSP@TD＋PDSP@Que給藥后DOX大多數(shù)都聚集在線粒體內(nèi)，細(xì)胞內(nèi)ROS含量明顯增加，較DOX、TD、PDSP@TD對(duì)線粒體膜電位的破壞作用更強(qiáng)（P＜0.05、0.01、0.001）。結(jié)論 PDSP@TD＋PDSP@Que相較于游離藥物具有較好的生物安全性及抗腫瘤活性，能較好的逆轉(zhuǎn)MCF-7/ADR細(xì)胞對(duì)DOX的耐藥，且具有較好的線粒體靶向性。

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

河南省科技攻關(guān)項(xiàng)目（232102311178）;河南省高等學(xué)校重點(diǎn)科研項(xiàng)目（25B350003）;河南應(yīng)用技術(shù)職業(yè)學(xué)院名師工作室項(xiàng)目（2022-03）;河南應(yīng)用技術(shù)職業(yè)學(xué)院教師創(chuàng)新團(tuán)隊(duì)項(xiàng)目（2022-02）