穿膜肽修饰葫芦素B纳米结构脂质载体的制备及体外评价  

Preparation of cell-penetrating peptide conjugated cucurbitaicin B loaded nanostructured lipid carriers and evaluation of cell membrane penetration effects in vitro

作　　者：金珊珊[1,2] 马晓星[2] 隋小宇[2] 刘畅[2] 张军威 韩翠艳[2] JIN Shanshan;MA Xiaoxing;SUI Xiaoyu;LIU Chang;ZHANG Junwei;HAN Cuiyan(School of Pharmacy, Jiamusi University, Jiamusi 154000, China;School of Pharmacy, Qiqihaer Medical University, Qiqihar 161006, China)

机构地区：[1]佳木斯大学药学院,黑龙江佳木斯154000 [2]齐齐哈尔医学院药学院,黑龙江齐齐哈尔161006

出　　处：《沈阳药科大学学报》2018年第4期253-258,288,共7页Journal of Shenyang Pharmaceutical University

基　　金：黑龙江省教育厅面上资助项目(12531787)

摘　　要：目的构建穿膜肽R8修饰葫芦素B纳米结构脂质载体(R8 conjugated cucurbitaicin B loaded nanostructured lipid carriers,R8-CuB-NLCs),并评价其体外穿膜效果。方法以SA-R8为穿膜材料,采用熔融乳化-低温固化法制备R8-CuB-NLCs,并表征;以香豆素-6(coumarin-6,Cou-6)为荧光探针、乳腺癌MCF-7为模型细胞,考察R8-CuB-NLCs的体外摄取行为,并采用CCK-8法评价细胞毒性。结果 R8-CuB-NLCs呈类圆形,平均粒径(35.00±1.20)nm,包封率(78.22±3.64)%,9 h内释放(76.90±3.05)%,具有缓释作用;穿膜肽R8修饰载香豆素-6纳米结构脂质载体(R8-Cou6-NLCs)的体外摄取率是普通载香豆素-6纳米结构脂质载体(Cou6-NLCs)的1.68倍(P<0.01);R8-CuBNLCs细胞毒性明显高于普通载葫芦素B纳米结构脂质载体(CuB-NLCs)和CuB溶液(P<0.01)。结论穿膜肽R8可以显著增强纳米粒的跨膜行为,提高CuB的药效。Objective To construct R8 conjugated cucurbitaicin B loaded nanostructured lipid carriers（ R8-CuB-NLCs） and evaluate its cell penetrating ability. Methods R8-CuB-NLCs w ere prepared by meltemulsification and low temperature-solidification method using penetrating material SA-R8 and characterized by morphology,average particle size and encapsulation efficiency. With coumarin-6（ Cou6） as fluorescent probe and M CF-7 as cell model,cellular uptake study and CCK-8 w as carried out to further investigate its cell penetration capacity and cell toxicity. Results Transmission electron microscopy（ TEM） show ed that the particles w ere irregular round like. The average particle size and encapsulation efficiency w ere（ 35. 00 ±1. 20） nm and（ 78. 22 ± 3. 64） % respectively. The cumulative release of CuB from R8-CuB-NLCs in 9 hours reached（ 76. 90 ± 3. 05） %. The cellular uptake efficiency of R8 conjugated Cou6 loaded nanostructured lipid carriers（ R8-Cou6-NLCs） by M CF-7 w as 1. 68 times over Cou6-NLCs. The cell toxicity of R8-CuB-NLCs on M CF-7 w as obviously higher than CuB-NLCs and CuB solution. Conclusions Cell penetrating peptide R8 decorated on CuB-NLCs could enhance the membrane penetration behavior of nanoparticles and cytology-inhibition for breast cancer w as improved.

关 键 词：穿膜肽R8 葫芦素B 纳米结构脂质载体 乳腺癌 MCF-7 

分 类 号：R94[医药卫生—药剂学]