白头翁皂苷B4和PD-L1siRNA共递送cRGD修饰靶向脂质体的制备及体外细胞摄取研究 点击下载
| 论文标题： | 白头翁皂苷B4和PD-L1siRNA共递送cRGD修饰靶向脂质体的制备及体外细胞摄取研究 |
| 英文标题： | |
| 中文摘要： | 目的 制备白头翁皂苷B4（AB4）和程序性死亡配体1（PD-L1）小干扰RNA（siP）共递送环精氨酰甘氨酸天冬氨酸序列（cRGD）修饰靶向脂质体（AB4/siP-c-L），并考察其体外细胞摄取行为。方法采用乙醇注入法制备cRGD修饰的AB4靶向脂质体（AB4-c-L），将AB4-c-L与20nmol/L的siP等体积混合，通过静电吸附得到AB4/siP-c-L。分别采用激光散射粒径测定仪、透射电子显微镜、超滤离心法、透析法、琼脂糖凝胶电泳法考察AB4/siP-c-L的粒径、Zeta电位、形态、包封率、药物含量、体外释放行为、血清稳定性。分别采用流式细胞术和共聚焦激光扫描技术评价小鼠Lewis肺癌细胞LLC对AB4/siP-c-L的摄取及其在细胞内的分布情况。结果AB4/siP-c-L的平均粒径为（187.4±3.1）nm，Zeta电位为（33.5±1.4）mV，形态呈类球形，AB4的包封率和含量分别为（95.2±0.4）%、（1.0±0.2）mg/mL；AB4/siP-c-L可较好地包裹siP，并具有较好的血清稳定性、pH敏感性以及缓释特性。LLC细胞对AB4/siP-c-L的摄取率显著高于游离药物，并能实现细胞质内富集。结论AB4/siP-c-L可有效实现AB4与基因药物siP共载，且对LLC细胞具有一定的体外靶向性。 |
| 英文摘要： | OBJECTIVE To prepare anemoside B4 （AB4） and programmed cell death ligand 1 （PDL1） siRNA （siP） co- delivered cRGD-modified targeting liposomes （AB4/siP-c-L）， and to study the cellular uptake in vitro. METHODS The cRGD- modified AB4-loaded targeted liposomes （AB4-c-L） were prepared by ethanol injection. AB4-c-L was mixed with 20 nmol/L siP in the same volume and AB4/siP-c-L was obtained through electrostatic adsorption. The particle size， Zeta potential， morphology， encapsulation efficiency and drug content， in vitro release behavior and serum stability of AB4/siP-c-L were investigated by laser scattering particle size tester， transmission electron microscopy， ultrafiltration centrifugation， dialysis and agar-gel electrophoresis block test. Cellular uptake of AB4/siP-c-L by Lewis lung cancer cells LLC and its intracellular localization were evaluated by flow cytometry and confocal laser scan technique. RESULTS The average particle size of AB4/siP-c-L was （187.4±3.1） nm， and the Zeta potential was （33.5±1.4） mV. AB4/siP-c-L was spheroidal in shape. The encapsulation efficiency and content of AB4 were （95.2±0.4） % and （1.0±0.2） mg/mL， respectively. AB4/siP-c-L could better package siP， and exhibited good serum stability， obvious pH sensitivity and sustained release property. The uptake rate of AB4/siP-c-L by LLC cells was significantly higher than that of free drug， and was able to accumulate in cytoplasm. CONCLUSIONS AB4/siP-c-L can effectively realize the co-loading of AB4 and gene drug siP， which has certain in vitro targeting to LLC cells. |
| 期刊： | 2023年第34卷第01期 |
| 作者： | 万安平；张婧；周雄；冯育林；刘骏；何瑶；李翔 |
| 英文作者： | WAN Anping，ZHANG Jing，ZHOU Xiong，FENG Yulin，LIU Jun，HE Yao，LI Xiang |
| 关键字： | 白头翁皂苷B4；程序性死亡配体1；小干扰RNA；脂质体；细胞摄取 |
| KEYWORDS： | anemoside B4； PD-L1； siRNA； liposome； cellular uptake |
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