目的 考察香菇多糖–番荔素纳米粒的性能，及其对小鼠黑色素瘤肺转移癌的体内外抑制效果。方法 采用反溶剂沉淀法制备香菇多糖–番荔素纳米粒，以动态光散射法测定粒径、分散系数（PDI）及Zeta电位，及其在不同生理介质（5%的葡萄糖、生理盐水、PBS的混悬液）中的稳定性；采用透射电子显微镜观察纳米粒的形态、大小；精确称量香菇多糖–番荔素纳米粒质量，并采用HPLC法测量番荔素中番荔辛，计算载药量；用酶标仪在540 nm处测量不同浓度（2、1.5、1、0.5、0.25、0.125 mg/mL）香菇多糖–番荔素纳米粒与5%葡萄糖溶液等渗液的吸光度，并计算溶血率；采用透析袋法考察香菇多糖–番荔素纳米粒的体外释放行为。用划痕实验与MTT实验对香菇多糖–番荔素纳米粒进行体外药效学考察。构建黑色素瘤肺转移癌小鼠模型，以紫杉醇注射液为阳性对照，对不同剂量香菇多糖–番荔素纳米粒进行体内药效学研究。结果 香菇多糖–番荔素纳米粒的粒径为（160.6±1.0）nm，PDI为0.082±0.023，Zeta电位为（－28.10±1.14）mV，透射电镜下呈球状。香菇多糖–番荔素纳米粒在5%的葡萄糖、血浆中稳定，无溶血现象；在体外可持续缓慢释放。体外研究结果显示，与番荔素原料药相比，香菇多糖–番荔素纳米粒对黑色素瘤B16F10细胞的迁移抑制作用及细胞毒性显著增加。体内药效学结果显示，香菇多糖–番荔素纳米粒iv给药14 d后，香菇多糖–番荔素纳米粒0.4 mg/mL组对黑色素瘤肺部转移抑制率可达到91.6%，S-100蛋白的表达也较模型组明显下调。结论 香菇多糖可作为稳定剂制备香菇多糖–番荔素纳米粒，香菇多糖–番荔素纳米粒对黑色素瘤肺转移癌初期具有显著的抑制作用。

Objective To investigate the performance of lentinan-annonaceous aceyogenins nanoparticles and their inhibitory effect on lung metastasis of mouse melanoma in vitro and in vivo. Methods Lentinan-annonaceous aceyogenins nanoparticles was prepared by antisolvent precipitation method. The particle size, dispersion coefficient (PDI) and Zeta potential of lentinan-annonaceous aceyogenins nanoparticles were determined by dynamic light scattering method, and their stability in different physiological media (5% glucose, 0.9% saline and PBS suspension) was determined. The morphology and size of the nanoparticles were observed by transmission electron microscopy. The mass of lentinan-annonaceous aceyogenins nanoparticles was accurately weighed, and the content of Squamoci in annonaceous aceyogenins was measured by HPLC, and the drug load was calculated. The absorbance of lentinan-annonaceous aceyogenins nanoparticles with different concentrations (2, 1.5, 1, 0.5, 0.25, and 0.125 mg/mL) and 5% glucose solution was measured by microplate reader at 540 nm, and the hemolysis rate was calculated. The release behavior of lentinan-annonaceous aceyogenins nanoparticles in vitro was investigated by dialysis bag method. In vitro pharmacodynamics of lentinan-annonaceous aceyogenins nanoparticles were investigated by scratch test and MTT test. A mice model of lung metastasis of melanoma was established, and Paclitaxel Injection was used as a positive control to study the pharmacodynamics of different doses of lentinan-annonaceous aceyogenins nanoparticles in vivo. Results The particle size of nanoparticles was (160.6 ±1.0) nm, PDI was (0.082 ±0.023), Zeta potential was (-28.10 ±1.14) mV, and the particle size was spherical under transmission electron microscope. Lentinan-annonaceous aceyogenins nanoparticles were stable in 5% glucose and plasma without hemolysis. Sustained slow release in vitro. The results of in vitro study showed that lentinan-annonaceous aceyogenins nanoparticles significantly increased the migration inhibition and cytotoxicity of melanoma B16F10 cells compared with annonaceous aceyogenins active pharmaceutical ingredients. In vivo pharmacodynamic results showed that 14 days after iv administration, the inhibition rate of melanoma lung metastasis in 0.4 mg/mL lentinan-annonaceous aceyogenins nanoparticles group was 91.6%, and the expression of S-100 protein was also significantly down-regulated compared with the model group. Conclusion Lentinan can be used as a stabilizer to prepare annonaceous aceyogenins nanoparticles, and lentinan-annonaceous aceyogenins nanoparticles had a significant inhibitory effect on melanoma lung metastatic carcinoma at the initial stage.

R944;R966

哈尔滨市应用技术研究与开发项目（2017RAQXJ115）