载山柰酚长循环靶向脂质体的处方筛选及抗卵巢癌疗效评估  

Formulation screening and efficacy evaluation against ovarian cancer of Kaempferol-loaded targeted liposomes

作　　者：陆主航 刘祖浩 王雪莲 唐俊 孟燕 常聪 LU Zhuhang;LIU Zuhao;WANG Xuelian;TANG Jun;MENG Yan;CHANG Cong(College of Pharmacy,Hubei University of Chinese Medicine,Hubei Wuhan 430065,China)

机构地区：[1]湖北中医药大学药学院,湖北武汉430065

出　　处：《中国医院药学杂志》2023年第21期2411-2418,共8页Chinese Journal of Hospital Pharmacy

基　　金：湖北中医药大学杰出青年基金(编号:2022zzxj006)。

摘　　要：目的:筛选载山柰酚长循环靶向脂质体(c-Lipo/KPF)的最佳处方工艺,并初步研究其抗卵巢癌疗效和机制。方法:以综合评分为指标,用正交试验法筛选c-Lipo/KPF的最优处方;用透射电镜、激光粒度仪、动态透析法、可见分光光度法等检测c-Lipo/KPF的形态、粒径、体外释药行为、血清稳定性和血液相容性;用MTT法、荧光显微镜和流式细胞术等检测空白脂质体的细胞相容性、c-Lipo/KPF对卵巢癌细胞的靶向效率和细胞毒性;用活性氧(ROS)检测试剂盒、凋亡检测试剂盒、周期检测试剂盒考察c-Lipo/KPF对卵巢癌细胞内ROS水平、凋亡和周期分布的影响。结果:c-Lipo/KPF的最佳处方工艺为磷脂、胆固醇、药物的摩尔比为80∶5∶4,超声时间为8 min;其粒径、Zeta电位、包封率(EE)和载药量(DL)分别为129 nm,-29 mV,95.2%和1.7%;溶血率低于5%;和普通脂质体(Lipo/KPF)相比,其血清稳定性、对A2780细胞的靶向效率和细胞毒性、升高细胞内ROS水平、诱导细胞G2/M期阻滞和凋亡的能力均显著提高。结论:按最佳处方工艺制备的c-Lipo/KPF,粒径均一,EE与DL较高,有良好的生物相容性、血清稳定性和卵巢癌细胞靶向效率,可以增加细胞内ROS水平、诱导G2/M期周期阻滞和凋亡、抑制细胞增殖。OBJECTIVE To screen the optimal formulation and preparation technology of Kaempferol-loaded long cycle targeted liposomes(c-Lipo/KPF),and to preliminarily study the anti-ovarian cancer efficacy and mechanism of the liposomes.METHODS The optimal formulation of c-Lipo/KPF was screened by orthogonal test with comprehensive score as the index;the morphology,particle size,drug release behavior in vitro,serum stability and blood compatibility of c-Lipo/KPF were detected respectively by transmission electron microscopy,laser particle size analyzer,dynamic dialysis and visible spectrophotometry;the cytocompatibility,targeting efficiency and cytotoxicity of c-Lipo/KPF against ovarian cancer cells were determined by MTT assay,fluorescence microscopy and flow cytometry,respectively.The effects of c-Lipo/KPF on ROS levels,apoptosis and cycle distribution of ovarian cancer cells were investigated respectively by reactive oxygen species(ROS)detection kit,apoptosis detection kit and cycle detection kit.RESULTS The optimal formulation and preparation parameter of c-Lipo/KPF were as follows:the molar ratio of phospholipid,cholesterol and drug was 80∶5∶4,and the ultrasonic time was 8 min;the particle size,Zeta potential,encapsulation rate(EE)and drug loading(DL)were 129 nm,-29 mV,95.2% and 1.7%,respectively;the hemolysis rate was less than 5%;compared with the ordinary liposomes(Lipo/KPF),many indicators of c-Lipo/KPF were improved,such as the serum stability,targeting efficiency and cytotoxicity to A2780 cells,increasing intracellular ROS level,and inducing G2/M phase arrest and apoptosis rate.CONCLUSION c-Lipo/KPF prepared according to the optimal formulation and preparation technology shows many advantages,such as uniform particle size,high EE and DL,good biocompatibility,serum stability and targeting efficiency to ovarian cancer cells.It can increase intracellular ROS level,induce G2/M phase cycle arrest and apoptosis,and inhibit cell proliferation.

关 键 词：山柰酚 长循环靶向脂质体 处方筛选 抗卵巢癌 

分 类 号：R944.1[医药卫生—药剂学]