外泌体装载蛇床子素的方法探究  

Study on method of osthole loaded by exosomes

作　　者：李明莉 胡冬生 杨秀竹[1] 王西墨[1] 鲍会静[1] LI Ming-li;HU Dong-sheng;YANG Xiu-zhu;WANG Xi-mo;BAO Hui-jing(Precision Medicine Laboratory,Tianjin Medical University Nankai Hospital,Tianjin 300100,China)

机构地区：[1]天津医科大学南开医院精准医学实验室,天津300100

出　　处：《中国药理学通报》2023年第10期1994-1998,共5页Chinese Pharmacological Bulletin

基　　金：天津市教委自然科学基金(No 2018KJ077)。

摘　　要：目的构建外泌体装载蛇床子素的载药体系。方法通过文献报道了解蛇床子素可以抑制卵巢癌细胞的增殖,用80μmol·L-1蛇床子素处理SKOV3细胞48 h,CCK-8实验进行验证;外泌体提取试剂盒提取外泌体,应用透射电子显微镜、纳米颗粒跟踪分析鉴定外泌体的表征;通过外泌体与蛇床子素共孵育2 h或4 h,确定最佳的载药条件,利用高效液相色谱仪测定其载药效率;最后用流式细胞术确定蛇床子素外泌体中的外泌体。结果蛇床子素可以抑制卵巢癌细胞SKOV3的增殖;成功提取外泌体,外泌体直径在100~200 nm,形态呈杯状;共孵育2 h和4 h的载药效率分别为0.76%、0.12%。结论成功构建外泌体装载蛇床子素的载药体系,并确定共孵育2 h为最佳的载药条件,载药效率为0.76%。Aim To construct a drug delivery system of osthole loaded by exosomes.Methods Osthole could inhibit the proliferation of ovarian cancer cells by literature.SKOV3 cells were treated with 80μmol·L-1 osthole for 48 h,and CCK-8 assay was used to verify the effect.The exosomes were extracted by exosome extraction kit,and the characteristics of exosomes were identified by transmission electron microscope(TEM)and nanoparticle tracking analysis(NTA).The exosomes were incubated with osthole for 2 or 4 hours to determine the optimal drug loading conditions,and the drug loading efficiency was measured by high performance liquid chromatography(HPLC).The presence of exosomes in osthole-Exos was determined by flow cytometry.Results Osthole could inhibit the proliferation of SKOV3 cells.Exosomes were successfully extracted.The diameter of exosomes was about 100 to 200 nm,and the shape was cup-shaped.The drug loading efficiency of 2 hours and 4 hours of co-incubation was 0.76%and 0.12%,respectively.Conclusions The osthole loaded exosome drug delivery system is successfully constructed,and co-incubation for 2 hours is determined as the optimal drug delivery condition with a drug delivery efficiency of 0.76%.

关 键 词：卵巢癌 外泌体 蛇床子素 载药方式 共孵育 载药效率 

分 类 号：R284.1[医药卫生—中药学] R329.2[医药卫生—中医学] R737.31