文献：Targeted combination therapy for glioblastoma by co-delivery of doxorubicin, YAP-siRNA and gold nanorods

文献链接：https://xueshu.baidu.com/usercenter/paper/show?paperid=12760pc0hx1c0030gx560e0002069257&site=xueshu_se

作者：Lihuang Lia,Yanxiu Liub,Jun Yangc,Qiang Zhanga , Mindan Lub , Xiumin Wangb,Liangcheng Lib,and Lei Ren

相关产品：DSPE-PEG2000-MAL 磷脂-聚乙二醇2000-马来亚酰胺

原文摘要：The combination of brain targeting drug delivery systems and multi-modalintervention pose a promising therapeutic approach for glioblastoma therapy. In this study, we developed an angiopep-2 peptide modified cationic liposomes loaded with doxorubicin, YAP-siRNA and gold nanorods (D/R@Ang2-Lip+Au) simultaneously, which has been characterized by high encapsulating efficiency for doxorubicin (95.4%) and effective binding of siRNA at N/P ratio of 20:1. The fluorescence imaging and flow cytometry analysis revealed high cellular uptake of

D/R@Ang2-Lip+Au. Real-time quantitative polymerase chain reaction and western blot analysis indicated that D/R@Ang2-Lip+Au could effectively inhibit the expression of YAP protein. In vitro and in vivo studies showed that D/R@Ang2-Lip+Au had the ability to target glioma cells, and achieved better anti-proliferative effects compared with non-targeted D/R@Lip+Au. Moreover, in

vivo experiment demonstrated that D/R@Ang2-Lip+Au was able to cross the blood-brain barrier, and combination therapy could significantly inhibit tumor growth. Therefore, the multifunctional D/R@Ang2-Lip+Au might provide a novel approach for effectively delivery of DOX, AuNRs and siRNA into the glioblastoma cells simultaneously and exerting synergistic therapeutic effects.

DSPE-PEG2000-MAL是一种功能化的磷脂-聚乙二醇衍生物。DSPE是一种常见的磷脂成分，赋予分子两亲性，使其能够在水性环境中自组装形成胶束或与细胞膜相互作用。中间的 PEG2000 链段具有高度的亲水性和良好的柔韧性，可有效改善分子的水溶性，并减少在生物体内的非特异性吸附。末端的马来酰亚胺（MAL）基团则具有很高的化学反应活性，能与含有巯基（- SH）的生物分子（如蛋白质、多肽）发生特异性的 Michael 加成反应。这种化合物在化合物递送系统中可用于制备脂质体，将化合物包裹在内部实现靶向运输；在生物标记领域，可通过与目标生物分子结合，实现对生物分子的标记和追踪。引用的这篇文献研究了一种同时装载阿霉素、YAP-siRNA和金纳米棒的多肽修饰阳离子脂质体（D/R@Ang2-Lip+Au），过程如下：

图：实验染色图形

用薄膜水化法制备阳离子脂质体

将脂质，包括DOTAP、DOPE、HSPC和Chol以一定摩尔比用氯仿溶解，在真空下旋转蒸发形成薄膜，然后用（NH4）2SO4水合，之后，用超声波产生均匀的单层脂质体。最后，用插入后技术制备了Blood vessels蛋白-2修饰的阳离子脂质体。即将DSPE-PEG2000-MAL加入脂质体中，孵育。

制备aunr和DOX共载脂质体（DOX@Lip+Au）

通过脂质体或脂质体-Blood vessels蛋白孵育，Aunr与脂质体（Lip+Au）复合物。一定重量比的Lip+Au和DOX孵育，通过透析膜分离游离DOX。对于AuNR、DOX和siRNA共载脂质体（D/R@Lip+Au），DOX@Lip+Au与siRNA以N/P混合。然后，室温孵育，以确保siRNA有效加载。

图：用靶向脂质体配方孵育的U87MG细胞的相对细胞活力。

结论：体外细胞摄取实验表明，DSPE-PEG2000-MAL参与制备的D/R@Ang2-Lip+Au可以同时将DOX和siRNA有效地传递到U87MG细胞中。此外，与非靶向D/R@Lip+Au相比，靶向配体Blood vessels蛋白-2的存在增加了细胞对D/R@Ang2-Lip+Au的摄取。此外，D/R@Ang2-Lip+Au在U87MG细胞中诱导的基因沉默效率高于D/R@Lip+Au。