Targeted intracellular delivery of resveratrol to glioblastoma cells using apolipoprotein E-containing reconstituted HDL as a nanovehicle.
Targeted intracellular delivery of resveratrol to glioblastoma cells using apolipoprotein E-containing reconstituted HDL as a nanovehicle.
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DOI：
10.1371/journal.pone.0135130
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Narayanaswami V
中科院分区：
文献类型：
--
作者：
Kim SH;Adhikari BB;Cruz S;Schramm MP;Vinson JA;Narayanaswami V
本研究的目的是利用载脂蛋白E3（apoE 3）将白藜芦醇转运到细胞内位点。重组高密度脂蛋白（rHDL）轴承白藜芦醇（rHDL/res）的制备使用磷脂和低密度脂蛋白受体（LDLr）的apoE 3结合域。生物物理特性表明，白藜芦醇被分配到磷脂双层的盘状rHDL/res颗粒（约19 nm直径）。免疫共沉淀研究表明，保留了apoE 3的LDLR结合能力。在胶质母细胞瘤A-172细胞中，通过直接荧光使用化学合成的NBD标记的白藜芦醇（res/NBD）嵌入rHDL/res。竞争和抑制研究表明，白藜芦醇的摄取是通过受体介导的内吞作用通过LDLr，与晚期内体/溶酶体中的apoE 3和res/NBD的共定位。我们建议，rHDL提供了一个理想的疏水环境，螯合白藜芦醇和含有载脂蛋白E3的rHDL作为一个有效的“纳米载体”，运输和提供白藜芦醇的靶向细胞内位点。
The objective of this study is to transport and deliver resveratrol to intracellular sites using apolipoprotein E3 (apoE3). Reconstituted high-density lipoprotein (rHDL) bearing resveratrol (rHDL/res) was prepared using phospholipids and the low-density lipoprotein receptor (LDLr)-binding domain of apoE3. Biophysical characterization revealed that resveratrol was partitioned into the phospholipid bilayer of discoidal rHDL/res particles (~19 nm diameter). Co-immunoprecipitation studies indicated that the LDLr-binding ability of apoE3 was retained. Cellular uptake of resveratrol to intracellular sites was evaluated in glioblastoma A-172 cells by direct fluorescence using chemically synthesized NBD-labeled resveratrol (res/NBD) embedded in rHDL/res. Competition and inhibition studies indicate that the uptake is by receptor mediated endocytosis via the LDLr, with co-localization of apoE3 and res/NBD in late endosomes/lysosomes. We propose that rHDL provides an ideal hydrophobic milieu to sequester resveratrol and that rHDL containing apoE3 serves as an effective “nanovehicle” to transport and deliver resveratrol to targeted intracellular sites.