目的 考察香菇多糖–番荔素納米粒的性能，及其對小鼠黑色素瘤肺轉(zhuǎn)移癌的體內(nèi)外抑制效果。方法 采用反溶劑沉淀法制備香菇多糖–番荔素納米粒，以動態(tài)光散射法測定粒徑、分散系數(shù)（PDI）及Zeta電位，及其在不同生理介質(zhì)（5%的葡萄糖、生理鹽水、PBS的混懸液）中的穩(wěn)定性；采用透射電子顯微鏡觀察納米粒的形態(tài)、大小；精確稱量香菇多糖–番荔素納米粒質(zhì)量，并采用HPLC法測量番荔素中番荔辛，計(jì)算載藥量；用酶標(biāo)儀在540 nm處測量不同濃度（2、1.5、1、0.5、0.25、0.125 mg/mL）香菇多糖–番荔素納米粒與5%葡萄糖溶液等滲液的吸光度，并計(jì)算溶血率；采用透析袋法考察香菇多糖–番荔素納米粒的體外釋放行為。用劃痕實(shí)驗(yàn)與MTT實(shí)驗(yàn)對香菇多糖–番荔素納米粒進(jìn)行體外藥效學(xué)考察。構(gòu)建黑色素瘤肺轉(zhuǎn)移癌小鼠模型，以紫杉醇注射液為陽性對照，對不同劑量香菇多糖–番荔素納米粒進(jìn)行體內(nèi)藥效學(xué)研究。結(jié)果 香菇多糖–番荔素納米粒的粒徑為（160.6±1.0）nm，PDI為0.082±0.023，Zeta電位為（－28.10±1.14）mV，透射電鏡下呈球狀。香菇多糖–番荔素納米粒在5%的葡萄糖、血漿中穩(wěn)定，無溶血現(xiàn)象；在體外可持續(xù)緩慢釋放。體外研究結(jié)果顯示，與番荔素原料藥相比，香菇多糖–番荔素納米粒對黑色素瘤B16F10細(xì)胞的遷移抑制作用及細(xì)胞毒性顯著增加。體內(nèi)藥效學(xué)結(jié)果顯示，香菇多糖–番荔素納米粒iv給藥14 d后，香菇多糖–番荔素納米粒0.4 mg/mL組對黑色素瘤肺部轉(zhuǎn)移抑制率可達(dá)到91.6%，S-100蛋白的表達(dá)也較模型組明顯下調(diào)。結(jié)論 香菇多糖可作為穩(wěn)定劑制備香菇多糖–番荔素納米粒，香菇多糖–番荔素納米粒對黑色素瘤肺轉(zhuǎn)移癌初期具有顯著的抑制作用。

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

哈爾濱市應(yīng)用技術(shù)研究與開發(fā)項(xiàng)目（2017RAQXJ115）