目的 制備山慈菇葡甘露聚糖（PBP）的硒納米粒，初步評價其體內(nèi)外抗腫瘤活性。方法 以原位生成的硒作為交聯(lián)劑，制備山慈菇葡甘露聚糖的納米粒（ PBP-SeNPs），使用單因素實驗和Box-Behnken響應(yīng)面法優(yōu)化PBP-SeNPs的處方及制備工藝，通過動態(tài)光散射法測定其平均粒徑、多分散性指數(shù)（ PDI）、ζ電位，透射電子顯微鏡（ TEM）觀察其外觀形態(tài)，同時考察納米粒在生理介質(zhì)及室溫放置下的粒徑變化。采用MTT法評估PBP-SeNPs對4T1細(xì)胞的體外毒性，并通過4T1荷瘤小鼠模型研究其體內(nèi)抗腫瘤效果。結(jié)果 最優(yōu)處方及制備工藝為PBP、亞硒酸鈉（Na₂SeO₃）、抗壞血酸以質(zhì)量比為9.79∶ 1∶3投料，PBP的質(zhì)量濃度為5.4 mg·mL^-1，反應(yīng)溫度為24℃，反應(yīng)時間為2.7 h時，PBP-SeNPs的粒徑為（126.400±6.402） nm，PDI為0.197±0.021，ζ電位為（ -8.17±0.35） mV，含硒量1.2%，載藥量為55.0%； TEM觀察PBP-SeNPs呈均一的球形，PBP-SeNPs在0.9%氯化鈉溶液、PBS、5%葡萄糖溶液或小鼠血漿中孵育12 h粒徑?jīng)]有顯著增加，室溫放置15 d仍穩(wěn)定。體外細(xì)胞毒實驗證實PBP-SeNPs能顯著抑制4T1乳腺癌細(xì)胞的增殖，對4T1細(xì)胞的生長抑制作用比同濃度PBP更強(qiáng)[半數(shù)抑制濃度（IC₅₀），62.64 vs 189.10 μg·mL^-1]；體內(nèi)抗腫瘤實驗中，PBP-SeNPs對荷瘤小鼠的抑瘤率明顯優(yōu)于陽性藥阿霉素注射液（68.3% vs 33.6%，P＜0.05）和PBP溶液（68.3% vs 49.7%，P＜0.05），且小鼠的體質(zhì)量和臟器指數(shù)均未出現(xiàn)顯著下降（P＞0.05）。結(jié)論 成功制備穩(wěn)定性良好的PBP-SeNPs，較PBP溶液顯著提高體內(nèi)外抗腫瘤活性。

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

北京市自然科學(xué)基金非共識項目（F251029）；中國醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康創(chuàng)新工程（2021-I2M-1-071）