目的 制备山慈菇葡甘露聚糖（PBP）的硒纳米粒，初步评价其体内外抗肿瘤活性。方法 以原位生成的硒作为交联剂，制备山慈菇葡甘露聚糖的纳米粒（ PBP-SeNPs），使用单因素实验和Box-Behnken响应面法优化PBP-SeNPs的处方及制备工艺，通过动态光散射法测定其平均粒径、多分散性指数（ PDI）、ζ电位，透射电子显微镜（ TEM）观察其外观形态，同时考察纳米粒在生理介质及室温放置下的粒径变化。采用MTT法评估PBP-SeNPs对4T1细胞的体外毒性，并通过4T1荷瘤小鼠模型研究其体内抗肿瘤效果。结果 最优处方及制备工艺为PBP、亚硒酸钠（Na₂SeO₃）、抗坏血酸以质量比为9.79∶ 1∶3投料，PBP的质量浓度为5.4 mg·mL^-1，反应温度为24℃，反应时间为2.7 h时，PBP-SeNPs的粒径为（126.400±6.402） nm，PDI为0.197±0.021，ζ电位为（ -8.17±0.35） mV，含硒量1.2%，载药量为55.0%； TEM观察PBP-SeNPs呈均一的球形，PBP-SeNPs在0.9%氯化钠溶液、PBS、5%葡萄糖溶液或小鼠血浆中孵育12 h粒径没有显著增加，室温放置15 d仍稳定。体外细胞毒实验证实PBP-SeNPs能显著抑制4T1乳腺癌细胞的增殖，对4T1细胞的生长抑制作用比同浓度PBP更强[半数抑制浓度（IC₅₀），62.64 vs 189.10 μg·mL^-1]；体内抗肿瘤实验中，PBP-SeNPs对荷瘤小鼠的抑瘤率明显优于阳性药阿霉素注射液（68.3% vs 33.6%，P＜0.05）和PBP溶液（68.3% vs 49.7%，P＜0.05），且小鼠的体质量和脏器指数均未出现显著下降（P＞0.05）。结论 成功制备稳定性良好的PBP-SeNPs，较PBP溶液显著提高体内外抗肿瘤活性。

Objective To prepare Pleione bulbocodioides glucomannan (PBP)-selenium nanoparticles and preliminarily evaluate their anti-tumor activity in vitro and in vivo. Methods PBP nanoparticles (PBP-SeNPs) were synthesized using in situ-generated selenium as a crosslinker. The formulation and preparation process were optimized through single-factor experiments and Box-Behnken response surface methodology. The optimized PBP-SeNPs were characterized by dynamic light scattering (DLS) for average particle size, polydispersity index (PDI), and ζ potential, while their morphology was observed via transmission electron microscopy (TEM). Their particle size changes in physiological media and during the storage at room temperature were monitored for stability assessment. MTT assay was used to evaluate their in vitro cytotoxicity against 4T1 breast cancer cells, and the in vivo anti-tumor efficacy was investigated in 4T1 tumor-bearing mice. Results The optimal formulation was the feeding mass ratio of PBP-sodium selenite (Na₂SeO₃)-ascorbic acid being 9.79∶ 1∶ 3 with the PBP concentration of 5.4 mg·mL^-1, and the system was reacted at 24℃ for 2.7 h. The resultant PBPSeNPs showed a particle size of (126.400 ±6.402) nm, a PDI of (0.197 ±0.021), a ζ potential of (-8.17 ±0.35) mV, with ta selenium content of 1.2% and a drug loading content of 55.0%. Transmission electron microscopy (TEM) showed that PBP-SeNPs were uniform spherical. PBP-SeNPs were stable in normal saline, PBS, 5% glucose solution or plasma with no significant particle size enlargement within 12 h, and remained stable at room temperature for 15 days as well. In MTT assay, PBP-SeNPs demonstrated significantly stronger growth inhibition against 4T1 cells than free PBP (IC₅₀, 62.64 vs 189.10 μg·mL^-1). In the in vivo study, PBP-SeNPs exhibited higher tumor inhibition rate over doxorubicin injections ( 68.3% vs 33.6%, P < 0.05) and PBP solution (68.3% vs 49.7%, P < 0.05), without inducing obvious body weight loss or organ index alterations (P > 0.05). Conclusion PBP-SeNPs with good stability were successfully prepared and significantly improved their anti-tumor activity in vitro and in vivo compared with PBP solution.

R285.5

北京市自然科学基金非共识项目（F251029）；中国医学科学院医学与健康创新工程（2021-I2M-1-071）