目的 制备薯蓣皂苷元丝素蛋白纳米粒（diosgenin silk fibroin nanoparticles，Doi-SF-NPs），并进行理化性质、肿瘤靶向性和体内抗肿瘤作用评价。方法 单因素考察Doi-SF-NPs主要影响因素，采用Box-Behnken设计-响应面法（Box-Behnken design-response surface methodology，BBD-RSM）优化Doi-SF-NPs处方工艺。使用透射电子显微镜（transmission electron microscopy，TEM）和傅里叶变换红外光谱（Fourier transform infrared spectroscopy，FT-IR）法进行表征。考察Doi-SF-NPs在不同pH值磷酸盐缓冲液（PBS）中的释药行为。采用MTT法和Annexin V/PI双染法考察Doi-SF-NPs对人前列腺癌PC3细胞的生长抑制及促凋亡作用。建立前列腺癌荷瘤裸鼠模型，比较薯蓣皂苷元原料药和Doi-SF-NPs在肿瘤组织中质量浓度，采用相对摄取率和峰浓度比值考察Doi-SF-NPs肿瘤靶向性。将前列腺癌荷瘤裸鼠分为模型（生理盐水）组、阳性对照（顺铂，2.0 mg/kg）组、薯蓣皂苷元原料药（10 mg/kg）组及Doi-SF-NPs低、高剂量（5、10 mg/kg）组，比较Doi-SF-NPs体内抗肿瘤效果。评价Doi-SF-NPs安全性。结果 Doi-SF-NPs最佳处方：丝素蛋白与薯蓣皂苷元质量比为11.94∶1，有机相与水相体积比为5.99∶1，制备时间为2.75 h。Doi-SF-NPs平均包封率、载药量、粒径和ζ电位分别（84.07±0.70）%、（6.41± 0.05）%、（164.29±4.43）nm和（-23.01±1.17）mV。薯蓣皂苷元与丝素蛋白通过氢键络合作用形成Doi-SF-NPs。Doi-SF-NPs外貌呈类圆形纳米颗粒。Doi-SF-NPs在pH 5.5、6.0 PBS中累积释放度较高，具有明显的pH值敏感性。薯蓣皂苷元原料药和Doi-SF-NPs对PC3细胞半数抑制浓度（median inhibition concentration，IC₅₀）分别为72.44、55.68 μmol/L。Doi-SF-NPs比薯蓣皂苷元原料药更有效地诱导了PC3细胞凋亡。Doi-SF-NPs在肿瘤组织内相对摄取率为3.23，峰浓度比值为2.60，证明Doi-SF-NPs能明显增加肿瘤组织的蓄积量。Doi-SF-NPs（10 mg/kg）将薯蓣皂苷元原料药（10 mg/kg）抑瘤率由29.47%提高至66.18%。Doi-SF-NPs未对荷瘤裸鼠主要脏器产生影响，使用安全性高。结论 Doi-SF-NPs可提高薯蓣皂苷元肿瘤靶向性，显著提高了抗肿瘤药效，有望开发成抗肿瘤药物。

Objective To prepare diosgenin silk fibroin nanoparticles (Doi-SF-NPs) and evaluate their physicochemical properties, tumor targeting and in vivo anti-tumor effects . Methods The main influencing factors of Doi-SF-NPs were investigated by single factor experiments, and Box-Behnken design-response surface methodology (BBD-RSM) was employed to optimize prescription process of Doi-SF-NPs. Doi-SF-NPs was characterized by transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FT-IR). Drug release behavior of Doi-SF-NPs in phosphate buffer saline solution with different pH values was investigated. MTT method and Annexin V/PI double staining method were used to investigate the inhibitory and proapoptotic effects of Doi-SF-NPs on PC3 cells, respectively. The nude mouse of prostate cancer was established, compared the concentrations of diosgenin and Doi-SF-NPs in tumor tissues, and then evaluated the tumor targeting ability of Doi-SF-NPs using the relative uptake rate and ratio of peak concentration. The nude mouse model of prostate cancer was divided into model (normal saline) group, positive control (cisplatin, 2.0 mg/kg) group, diosgenin (10 mg/kg) group, low-dose Doi-SF-NPs (5 mg/kg) group and high-dose Doi-SF-NPs (10 mg/kg) group, the in vivo anti-tumor effects of Doi-SF-NPs was compared with diosgenin. Safety evaluation of Doi-SF-NPs was also performed. Results Optimal prescription process of Doi-SF-NPs nanoparticles as follows: The mass ratio of silk fibroin to diosgenin was 11.94:1, the volume ratio of the organic phase to aqueous phase was 5.99:1, and the preparation time was 2.75 h. Average envelopment efficiency, drug loading, particle size and ζ potential were (84.07 ± 0.70)%, (6.41 ± 0.05)%, (164.29 ± 4.43) nm and (−23.01 ± 1.17) mV, respectively. Diosgenin formed Doi-SF-NPs with silk fibroin through hydrogen bonding. The appearance of Doi-SF-NPs was spherical nanoparticles. Doi-SF-NPs existed higher cumulative release in phosphate buffer solution of pH 5.5 and 6.0, which demonstrated that Doi-SF-NPs had obvious pH-sensitivity. MTT method results showed that IC₅₀ of diosgenin and Doi-SF-NPs on PC3 cells were 72.44 and 55.68 μmol/L, respectively. Doi-SF-NPs more effectively induced apoptosis of PC3 cells than that of diosgenin. The relative uptake rate of Doi-SF-NPs in in tumor tissue was 3.23, and the peak concentration ratio was 2.60, which demonstrating that Doi-SF-NPs could significantly increase the accumulation in tumor tissue. Doi-SF-NPs (10 mg/kg) increased the tumor inhibition rate of diosgenin (10 mg/kg) from 29.47% to 66.18%. Doi-SF-NPs did not affect the major organs of tumor-bearing nude mouse, which showed that the safety of Doi-SF-NPs was high. Conclusion Doi-SF-NPs could improve the tumor targeting of diosgenin and significantly enhanced anti-tumor efficacy. Therefore, Doi-SF-NPs had the potential to be developed into an antitumor drug.

R283.6

2025年度河南省高等学校重点科研项目（25B310010）；2024年度河南应用技术职业学院校级科研课题（2024-KJ-69）；2025年度河南省科技攻关课题（252102310468）；2026年度河南省科技攻关课题（262102310551）