海南医科大学学报2025，Vol.31Issue(4)：241-247,253,8.DOI:10.13210/j.cnki.jhmu.20241101.002
人脐带间充质干细胞源外泌体负载刺槐素的制备方法研究
Research on preparation method of acacetin loaded on exosomes derived from human umbilical cord mesenchymal stem cells
摘要
Abstract
Objective:To construct exosomes(EXOs)drug delivery system of acacetin.Methods:Using exosomes derived from human umbilical cord mesenchymal stem cells(hUMSCs)as drug carriers,The exosomes loaded with acacetin(EX-Aca)were prepared by co-incubation,ultrasound and repeated freeze-thaw methods.The encapsulation rate and loading rate of EX-Aca were determined by high performance liquid chromatography.Transmission electron microscopy(TEM),particle size analysis and Western Blot were used to detect the characterization of EXOs and EX-Aca.The safety of Ex-Aca was tested by in vitro toxic-ity test,and its cellular uptake capacity was also investigated.Result:The results of high performance liquid chromatography showed that the encapsulation rate and drug loading rate of co-incubation method were higher than that of ultrasonic method and re-peated freeze-thaw method.When the concentration of acacetin was 80 μg/mL,the encapsulation rate and drug loading were the highest,which were 91.90%and 19.68%,respectively.Under transmission electron microscopy(TEM),EXOs and EX-Aca were cup-and-saucer-shaped,with average particle sizes of(102.356 96±0.656 23)nm and(117.300 38±0.747 83)nm,respec-tively,showing typical exosome characteristics.Western Blot detection of EXOs and EX-Aca surface markers CD9,ALIX and TSG101 protein expression.Therefore,the optimal preparation parameters of EX-Aca were as follows:The concentration of acac-etin was 80 μg/mL,and hUMSCs-derived exosomes were incubated at 4℃for 48 h.In vitro experiments confirmed that EX-Aca had low cytotoxicity,cell survival rate was>94%,and EX-Aca could be taken up by BV2 cells.Conclusion:The exosomes drug delivery system of acacetin was successfully constructed,and the prepared EX-Aca had low toxicity to cells and certain targeting.关键词
刺槐素/外泌体/药物载体/高效液相色谱/包封率Key words
Acacetin/Exosomes/Drug carrier/High performance liquid chromatography/Encapsulation efficiency分类
医药卫生引用本文复制引用
张胜男,周玲,叶勒丹·马汉,张晓玲,补娟..人脐带间充质干细胞源外泌体负载刺槐素的制备方法研究[J].海南医科大学学报,2025,31(4):241-247,253,8.基金项目
This study was supported by the Basic and Clinical Research of Stem Cell Biotherapy,People's Hospital of Xinjiang Uygur Autonomous Region(GXBZX-2023007) （GXBZX-2023007）
and Natural Science Foundation of Xinjiang Uygur Autonomous Region(2020D01C095)新疆维吾尔自治区人民医院干细胞生物治疗基础和临床研究专项(GXBZX-2023007) （2020D01C095）
新疆维吾尔自治区自然科学基金(2020D01C095) （2020D01C095）
