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目的 制备负载雷帕霉素的光敏纳米盘，并评价其体外抗卵巢癌活性。方法 采用薄膜水化法制备纳米盘，通过动态光散射和透射电子显微镜表征其粒径、电位和形貌；建立高效液相色谱法测定雷帕霉素含量，并验证方法专属性、线性、精密度、重复性、稳定性与加样回收率。通过单线态氧检测探针与反向透析法，分别评估了纳米盘在近红外光下的单线态氧生成与光控释药性能。通过检测细胞代谢活性，评估纳米盘对人卵巢癌细胞OVCAR-8的杀伤作用。采用共聚焦显微镜与流式细胞术，分别对纳米盘的细胞摄取行为及其内吞机制进行探究。结果 纳米盘粒径为（63.66±0.27）nm，电位为（-18.2±2.4）mV，稳定性良好。透射电镜结果显示其具有类球状或短棒状形态。雷帕霉素在0.2～40μg/mL的浓度范围内峰面积线性良好（R2=0.999 9），精密度、重复性和回收率的相对标准偏差均小于5%。在近红外光照射下，纳米盘可有效产生单线态氧；同时，单线态氧可诱导脂质膜氧化，进而显著促进雷帕霉素的响应性释放（P<0.001）。纳米盘本身无显著细胞毒性，而经近红外光照射后对卵巢癌细胞的毒性显著增强。细胞摄取实验证实，纳米盘主要通过网格蛋白介导的能量依赖型内吞途径进入细胞（P<0.001），光照后可触发药物释放。结论 成功制备雷帕霉素光敏纳米盘，其在体外表现出显著的光控释药特性及增强的抗卵巢癌活性。
Abstract：Objective To prepare photosensitive nanodiscs loaded with rapamycin and evaluate their in vitro antitumor activity against ovarian cancer.Methods Nanodiscs were prepared by the thin-film hydration method, and their particle size, potential and morphology were characterized by dynamic light scattering and transmission electron microscopy. High performance liquid chromatography was established for the determination of rapamycin content, and the specificity, linearity, precision, repeatability, stability, and recovery rate of the method were verified. The singlet oxygen generation and light-controlled drug release properties of nanodiscs under near-infrared light were evaluated respectively by singlet oxygen detection probe and reverse dialysis method. The cytotoxicity of the nanodiscs against human ovarian cancer cells(OVCAR-8) was evaluated by detecting cell metabolic activity. Confocal microscopy and flow cytometry were used to investigate the cellular uptake and endocytosis mechanisms of the nanodiscs.Results The nanodiscs exhibited a hydrodynamic diameter of(63. 66±0. 27) nm, a zeta potential of(-18. 2 ± 2. 4) mV, and demonstrated good stability. The results from transmission electron microscopy revealed that the nanodiscs exhibited spherical and short rod-like morphologies. The peak area of rapamycin showed good linearity in the concentration range of 0. 2-40 μg/mL(R2=0. 999 9), and the relative standard deviations of precision, repeatability and recovery were all less than 5%. Under near-infrared light irradiation, the nanodiscs could effectively produce singlet oxygen. At the same time, the singlet oxygen generated could induce lipid membrane oxidation, which significantly promoted the responsive release of rapamycin(P<0. 001). Nanodiscs had no obvious cytotoxicity, but the cytotoxicity to ovarian cancer cells was significantly enhanced after near-infrared irradiation. Cellular uptake experiments confirmed that the nanodiscs mainly entered cells through clathrin-mediated, energy-dependent endocytosis(P<0. 001), with drug release triggered by light exposure.Conclusion Rapamycin photosensitive nanodiscs were successfully prepared, which showed significant light-controlled release properties and enhanced anti-ovarian cancer activity in vitro.
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基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2026.06.004
中图分类号:R943
引用信息:
[1]张魁枝,卢光照,鲁莹,等.雷帕霉素光敏纳米盘的制备及体外抗卵巢癌活性评价[J].安徽医科大学学报,2026,61(06):994-1002.DOI:10.19405/j.cnki.issn1000-1492.2026.06.004.
基金信息:
上海市自然科学基金项目（编号：23ZR1477500）~~
2026-05-19
2026-05-19
2026-05-19