目的 制备活性氧（reactive oxygen species，ROS）响应的共载榄香烯（elemene，Ee）和二氢卟吩e6（chlorin e6，Ce6）脂质体（Ee-Ce6-L），对其理化性质进行表征，并研究Ee-Ce6-L对膀胱癌5637细胞的抑制作用。方法 采用MTT法筛选榄香烯与Ce6（Ee+Ce6）的协同比例，通过薄膜水化法制备Ee-Ce6-L，并对其形态、粒径、包封率和载药量、稳定性、ROS生成能力、体外释放度等性能进行表征；MTT实验考察Ee-Ce6-L对5637细胞的毒性；Annexin V-FITC/PI双染实验研究Ee-Ce6-L对细胞的凋亡作用；荧光显微镜及荧光酶标仪考察其对ROS生成能力及线粒体膜电位（mitochondrial membrane potential，MMP）的影响；通过免疫印迹法（Western blotting）考察凋亡通路相关蛋白的表达。结果 Ee+Ce6在40∶1～200∶1均具有协同作用，且在120∶1时协同效果最好；Ee-Ce6-L具有较好的稳定性和分散性，经660 nm激光照射后粒径由（194.27±0.20）nm变为（390.97±2.76）nm，ζ电位由（-19.06±0.07）mV变为（-6.30±0.34）mV；体外释放实验表明，Ee-Ce6-L具有缓释和光控释药的双重作用；榄香烯和Ce6的包封率分别为（94.02±0.38）%、（96.95±0.70）%，榄香烯和Ce6的载药量分别为（11.51±0.46）%、（2.32±0.01）%；Ee-Ce6-L有较强的单线态氧生成能力；体外细胞实验表明，Ee-Ce6-L能够通过增加ROS的产生和降低MMP，以及降低B细胞淋巴瘤-2（B-cell lymphoma-2，Bcl-2）蛋白表达，增加Bcl-2相关X蛋白（Bcl-2-associated X protein，Bax），裂解的多聚ADP核糖聚合酶（cleaved poly ADP-ribose polymerase，cleaved PARP）和裂解的半胱氨酸天冬氨酸蛋白酶-3（cleaved cystein-asparate protease-3，cleaved Caspase-3）蛋白的表达，促进5637细胞凋亡。结论 Ee-Ce6-L具有ROS响应释放特性，其能够通过中药-光疗作用协同抑制5637细胞增殖和诱导细胞凋亡。

Objective Reactive oxygen species (ROS)-responsive co-loaded elemene (Ee) and chlorin e6 (Ce6) liposomes (Ee-Ce6-L) were prepared, characterization of their physicochemical properties, and evaluated for their inhibitory effects on bladder cancer 5637 cells. Methods The synergistic ratio of elemene and Ce6 (Ee + Ce6) was screened using the MTT assay. Ee-Ce6-L was prepared via the thin-film hydration method and characterized for morphology, particle size, encapsulation efficiency, drug loading capacity, stability, ROS generation capacity, and in vitro release rate. MTT assays evaluated the cytotoxicity of Ee-Ce6-L against 5637 cells. Annexin V-FITC/PI double staining was used to study the apoptotic effects of Ee-Ce6-L on cells. Fluorescence microscopy and a fluorometer were employed to examine its impact on ROS generation capacity and mitochondrial membrane potential (MMP). Western blotting analysis was conducted to investigate the expression of apoptosis pathway-related proteins. Results Ee + Ce6 exhibited synergistic effects across the 40:1 to 200:1 concentration range, with optimal synergy observed at 120:1; Ee-Ce6-L exhibited good stability and dispersibility. After irradiation with 660 nm laser, the particle size changed from (194.27 ± 0.20) nm to (390.97 ± 2.76) nm, and the ζ potential shifted from (−19.06 ± 0.07) mV to (−6.30 ± 0.34) mV. In vitro release experiments demonstrated that Ee-Ce6-L exhibits dual functions of sustained release and light-controlled drug release; the encapsulation efficiencies of elemene and Ce6 were (94.02 ± 0.38)% and (96.95 ± 0.70)%, respectively, while the drug loading capacities of Ee and Ce6 were (11.51 ± 0.46)% and (2.32 ± 0.01)%; Ee-Ce6-L exhibited strong singlet oxygen generation capacity; In vitro cell experiments demonstrated that Ee-Ce6-L promoted apoptosis in 5637 cells by increasing ROS production and reducing MMP, as well as by decreasing B-cell lymphoma-2 (Bcl-2) protein expression while increasing Bcl-2-associated X protein (Bax), cleaved poly ADP-ribose polymerase (cleaved PARP), and cleaved cystein-asparate protease-3 (cleaved Caspase-3) protein expression. Conclusion Ee-Ce6-L exhibits ROS-responsive release properties and synergistically inhibits 5637 cell proliferation and induces apoptosis through the combined effects of traditional Chinese medicine and phototherapy.

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国家自然科学基金青年项目（82505363）；国家自然科学基金青年项目（82204935）；国家中医药管理局赵锋全国老药工传承工作室建设项目（国中医药人教函［2024］255号）；国家中医药管理局岳宝森全国老药工传承工作室建设项目（国中医药人教函［2025］181号）；陕西省教育厅重点科学研究计划项目（25JR058）；陕西中医药大学学科创新团队项目（2019-YL11）；西安市卫生健康委员会项目（2025qn01）