黄芩苷PEG-PE纳米胶束的制备、表征与细胞毒性研究  

Study on Preparation,Characterization and Cytotoxicity of Baicalin PEG-PE Nanomicelles

作　　者：宁国庆[1] 吴洁[1] 葛晨亮[1] 周定荣 唐义信 NING Guoqing;WU Jie;GE Chenliang;ZHOU Dingrong;TANG Yixin(The First Affiliated Hospital of Nanhua University,Hunan Hengyang 421001,China;Central Laboratory,Xiangya Second Hospital of Central South University,Changsha 410011,China)

机构地区：[1]南华大学附属第一医院心内科,湖南衡阳421001 [2]中南大学湘雅二医院中心实验室,长沙410011

出　　处：《中国药房》2019年第11期1487-1491,共5页China Pharmacy

基　　金：湖南省卫生计生委科研计划课题(No.20180059)

摘　　要：目的:制备黄芩苷(BAI)聚乙二醇衍生化磷脂酰乙醇胺(BAI@PEG-PE)纳米胶束,并对其进行表征和细胞毒性研究。方法:采用薄膜水化法制备BAI@PEG-PE 纳米胶束,观察其外观特征,检测其粒径、多分散系数(PDI)、Zeta 电位、载药量、包封率。比较BAI 原料药和BAI@PEG-PE 纳米胶束在pH 7.4 磷酸盐缓冲液中1~84 h 内的释药情况。以香豆素6 为荧光探针,观察PEG-PE纳米胶束在H9c2 心肌细胞中的分布。将H9c2 心肌细胞分为模型组、BAI原料药组和BAI@PEG-PE纳米胶束组,用不含药或含相应药物的无血清DMEM 培养液培养0.5 h 后,用异丙肾上腺素诱导心肌细胞凋亡,比较3 组细胞的细胞核形态变化、细胞凋亡率和凋亡相关蛋白B淋巴细胞瘤2(Bcl-2)及其X蛋白(Bax)的蛋白表达水平。结果:所制备的BAI@PEG-PE纳米胶束呈现大小比较均一的圆球形,其粒径为(16.7±0.8)nm,PDI 为0.11±0.01,Zeta 电位为(-18.4±0.6)mV,载药量为(7.84±0.65)%,包封率为(85.7±4.9)%(n=3),84 h 的累积释放度为76.5%,而BAI 原料药在24 h 内已基本释放完全。PEG-PE 纳米胶束可增强H9c2 心肌细胞对香豆素6 的摄取,且主要聚集在线粒体周围。与模型组比较,BAI 原料药组和BAI@PEG-PE纳米胶束组细胞的凋亡形态明显改善,凋亡率明显降低,Bcl-2 蛋白表达明显增强,Bax蛋白表达水平明显降低,差异均有统计学意义(P<0.05 或P<0.01),其中BAI@PEG-PE纳米胶束组的上述效果更明显(P<0.05 或P<0.01)。结论。成功制得BAI@PEG-PE纳米胶束,其具有明显的缓释作用、心肌靶向性,可预防心肌细胞的凋亡。OBJECTIVE:To prepare Baicalin-loaded Polyethylene glycol-derivatized phosphatidylethanolamine(BAI@PEG-PE) nanomicelles,and to characterize it and study its cytotoxicity. METHODS:BAI@PEG-PE nanomicelles were prepared by film hydration method and their appearance characteristics were observed. The particle size,polydispersity index,Zeta potential, drug-loading amount and encapsulation efficiency of the nanomicelles were detected. Drug release of BAI raw material and BAI@PEG-PE nanomicelles in pH 7.4 phosphate buffer were compared within 1-84 h. Using coumarin 6 as fluorescent probe,the distribution of PEG-PE nanomicelles in H9c2 cardiomyocytes were observed. H9c2 cardiomyocytes were divided into model group, BAI raw material group and BAI@PEG-PE nanomicelles group. After treated with serum-free DMEM medium containing no or corresponding drugs for 0.5 h,isoproterenol was used to induce cardiomyocyte apoptosis. Nuclear morphology,cell apoptosis rate and protein expression of Bcl-2 and Bax were compared with among 3 groups. RESULTS:Prepared BAI@PEG-PE nanomicelles were uniform globular shape. The particle size was(16.7±0.8)nm,PDI was 0.11±0.01 and Zeta-potential was(-18.4±0.6) mV;drug-loading amount was(7.84±0.65)%,encapsulation efficiency was(85.7±4.9)%(n=3). Accumulative release rate was 76.5% within 84 h. BAI raw material was released completely within 24 h. PEG-PE nanomicelles could strengthen the intake of coumarin 6 in H9c2 cardiomyocytes, mainly gathering around mitochondria. Compared with model group, the apoptosis morphology of cardiomyocytes were improved significantly in BAI raw material group and BAI@PEG-PE nanomicelles group;apoptosis rate was decreased significantly;protein expression of Bcl-2 was increased significantly;protein expression of Bax was decreased significantly with statistical significance(P<0.05 or P<0.01). Above effects of BAI@PEG-PE nanomicelles group were more significant(P<0.05 or P<0.01). CONCLUSIONS:BAI@PEG-PE nanomicelles are prepared successfully,and show significant sustained

关 键 词：聚乙二醇衍生化磷脂酰乙醇胺 纳米胶束 黄芩苷 体外释放 H9C2心肌细胞 

分 类 号：R943[医药卫生—药剂学]