首页|靶向载长春新碱超声微泡的制备及体外寻靶实验研究

靶向载长春新碱超声微泡的制备及体外寻靶实验研究

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目的 制备载长春新碱(VCR)且靶向结合外周神经髓鞘细胞的超声脂质微泡,观察其体外寻靶情况.方法 采用薄膜水化-机械振荡法制作载药超声微泡(VCR-MBs),生物素-亲和素桥接法制作靶向载药超声微泡(VCR-TMBs),观察 VCR-TMBs 形态、粒径、稳定性、体外显影、包封率及靶向性等理化性质.流式细胞仪检测VCR-TMBs对细胞的损伤及凋亡作用,体外通过VCR-TMBs联合超声(US)对大鼠肾动脉处理,HE染色观察脱髓鞘改变.结果 MBs、VCR-MBs 及 VCR-TMBs 光镜下均成圆形,分布均匀.MBs、VCR-MBs 及 VCR-TMBs 粒径分别为(3.22±1.07)、(3.48±1.06)及(3.22±0.79)μm,差异无统计学意义(P>0.05);与MBs、VCR-MBs相比,VCR-TMBs保存3d时浓度明显下降(P<0.05);采用高效液相色谱法测定 VCR-TMBs 的包封率及载药率分别为(82.90±9.98)%、(2.07±0.25)%;免疫荧光法测定VCR-TMBs可靶向聚集在Schwann细胞膜上;流式细胞仪检测示VCR-TMBs +US促细胞凋亡作用优于单纯VCR +US及VCR-MBs + US(P<0.05);组织HE染色示VCR-TMBs联合US可有效使体外大鼠肾动脉神经脱髓鞘及变性坏死.结论 成功制备VCR-TMBs,其可靶向识别Schwann细胞,促进细胞凋亡,使大鼠肾动脉神经脱髓鞘改变,可为后期体内VCR-TMBs实现肾动脉周围神经去交感化做前期准备.
Preparation and in Vitro Targeting Experiment of Vincristine-Loaded Ul-trasound Microbubbles
Objective To prepare vincristine(VCR)-loaded lipid ultrasound(US)microbubbles(VCR-MBs)tar-geting to peripheral nerve myelin cells,and to observe the in vitro targeting experiment.Methods VCR-MBs were prepared by thin film hydration and mechanical oscillation method,and targeted VCR-MBs(VCR-TMBs)was prepared by biotin-avidin bridging method.The morphology,particle size,stability,in vitro development,encapsulation rate and targeting properties of VCR-TMBs were observed.The effect of VCR-TMBs on cell injury and apoptosis was detected by flow cytometry.The rat renal artery was treated by VCR-TMBs combined with US in vitro,and demyelination was observed by HE staining.Results MBs,VCR-MBs and VCR-TMBs were all circular and evenly distributed under light microscope.The particle sizes of MBs,VCR-MBs and VCR-TMBs were(3.22±1.07),(3.48±1.06)and(3.22±0.79)μm,respectively,with no significant differ-ence(P>0.05).Compared with MBs and VCR-MBs,the concentration of VCR-TMBs decreased significantly after3 days of storage(P<0.05).The encapsulation rate and drug loading rate of VCR-TMBs determined by high-performance liquid chro-matography were(82.90±9.98)%and(2.07±0.25)%,respectively.VCR-TMBs was detected by immunofluorescence and could be targeted to accumulate on Schwann cell membrane.Flow cytometry showed that VCR-TMBS +US had better ap-optotic effect than VCR +US and VCR-MBS +US(P<0.05).HE staining showed that VCR-TMBs +US could effectively de-myelinate and denaturate the renal artery nerve in vitro.Conclusion The successful preparation of VCR-TMBs can target Schwann cells,promote cell apoptosis and demyelinate renal artery nerves of rats,which can be a preliminary preparation for the later realization of renal artery nerve dessympathization by VCR-TMBs in vivo.

VincristineTargeted drug-loaded ultrasound microbubblesTargeted recognitionSchwann cellsKid-ney desensitizationChromatography,high-performance liquidApoptosis

杨红、陈秋宏、黄娟、郭婧仪、王昱媛、吴奇

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610057 成都,成都医学院第二附属医院核工业四一六医院心血管内科

610500 成都,成都医学院

长春新碱 靶向载药超声微泡 靶向识别 Schwann细胞 肾去交感化 色谱法,高压液相 细胞凋亡

四川省中医药管理局科研项目成都市科技厅技术创新研发项目成都市医学重点专科

2023MS1772022-YF05-01459-SNCDS2022Z076

2024

10.3969/j.issn.2095-3097.2024.02.004

转化医学杂志
海军总医院

转化医学杂志

CSTPCD
影响因子：0.671
ISSN：2095-3097
年,卷(期)：2024.13(2)
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