Amphiphilic core-shell nanoparticles containing dense polyethyleneimine shells for efficient delivery of microRNA to Kupffer cells.
Amphiphilic core-shell nanoparticles containing dense polyethyleneimine shells for efficient delivery of microRNA to Kupffer cells.
复制标题
含有致密聚乙烯亚胺壳的两亲性核壳纳米粒子，可将 microRNA 有效递送至库普弗细胞
DOI：
10.2147/ijn.s101251
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发表时间：
2016
影响因子：
8
通讯作者：
Gong J
中科院分区：
文献类型：
--
作者：
Liu Z;Niu D;Zhang J;Zhang W;Yao Y;Li P;Gong J
将外源基因高效且有针对性地递送至巨噬细胞仍然是一个巨大的挑战。目前的基因递送方法在某些类型的细胞（尤其是库普弗细胞）体内往往导致细胞摄取效率低下。在本文中，我们证明了由明确的疏水性聚甲基丙烯酸甲酯（PMMA）核和支化聚乙烯亚胺（PEI）壳组成的两亲性核 - 壳纳米粒子（NPs）（表示为PEI@PMMA NPs）是将载有微小RNA（miRNA）的质粒递送至库普弗细胞的有效纳米载体。PEI@PMMA NPs的平均流体动力学直径为279 nm，尺寸分布较窄。这些纳米粒子在水中还具有高达 +30 mV的正表面电荷，从而能够有效凝聚带负电荷的质粒DNA。凝胶电泳分析表明，所得到的PEI@PMMA NPs能够在重量比为25:1（N/P比等于45:1）的情况下完全凝聚miRNA质粒。细胞计数试剂盒 - 8检测和流式细胞术结果表明，PEI@PMMA/miRNA NPs对库普弗细胞显示出低细胞毒性和细胞凋亡活性。48小时后，最大细胞转染效率达到34.7%，远高于使用商业脂质体转染试剂2000（1.7%）所获得的效率。生物透射电子显微镜观察显示，PEI@PMMA NPs主要分布在库普弗细胞的细胞质中。此外，与对照组相比，在体外和体内，靶核因子κB P65的蛋白表达均受到显著抑制（P<0.05）。这些结果表明，具有独特两亲性核 - 壳纳米结构的PEI@PMMA NPs是将miRNA质粒递送至库普弗细胞的有前景的纳米载体。
Efficient and targeted delivery approach to transfer exogenous genes into macrophages is still a great challenge. Current gene delivery methods often result in low cellular uptake efficiency in vivo in some types of cells, especially for the Kupffer cells (KCs). In this article, we demonstrate that amphiphilic core–shell nanoparticles (NPs) consisting of well-defined hydrophobic poly(methyl methacrylate) (PMMA) cores and branched polyethyleneimine (PEI) shells (denoted as PEI@PMMA NPs) are efficient nanocarriers to deliver microRNA (miRNA)-loaded plasmid to the KCs. Average hydrodynamic diameter of PEI@ PMMA NPs was 279 nm with a narrow size distribution. The NPs also possessed positive surface charges up to +30 mV in water, thus enabling effective condensation of negatively charged plasmid DNA. Gel electrophoresis assay showed that the resultant PEI@PMMA NPs were able to completely condense miRNA plasmid at a weight ratio of 25:1 (N/P ratio equal to 45:1). The Cell Counting Kit-8 assay and flow cytometry results showed that the PEI@PMMA/miRNA NPs displayed low cytotoxicity and cell apoptosis activity against the KCs. The maximum cell transfection efficiency reached 34.7% after 48 hours, which is much higher than that obtained by using the commercial Lipofectamine™ 2000 (1.7%). Bio-transmission electron microscope observation revealed that the PEI@PMMA NPs were mainly distributed in the cytoplasm of the KCs. Furthermore, when compared to the control groups, the protein expression of target nuclear factor κB P65 was considerably inhibited (P<0.05) both in vitro and in vivo. These results demonstrate that the PEI@PMMA NPs with a unique amphiphilic core–shell nanostructure are promising nanocarriers for delivering miRNA plasmid to KCs.