香菇多糖–番荔素纳米粒的性能考察及其对小鼠黑色素瘤肺转移癌的抑制作用  

Properties of lentinan-annonaceous aceyogenins nanoparticles and its inhibitory effect on lung metastasis of melanoma in mice

作　　者：高广宇 李春梅 GAO Guang-yu;LI Chun-mei(Harbin University of Commerce,Harbin 150076,China)

机构地区：[1]哈尔滨商业大学,黑龙江哈尔滨150076

出　　处：《现代药物与临床》2022年第9期1945-1952,共8页Drugs & Clinic

基　　金：哈尔滨市应用技术研究与开发项目(2017RAQXJ115)。

摘　　要：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 com

关 键 词：番荔素 香菇多糖 纳米粒 黑色素瘤 肺转移癌 S-100蛋白 

分 类 号：R944[医药卫生—药剂学] R966[医药卫生—药学]