Li Bo, Guo Wanrong, Zhang Fan, Liu Meiyan, Wang Shang, Liu Zhonghua, Xiang Shuanglin, Zeng Youlin
National & Local Joint Engineering Laboratory for New Petro-chemical Materials and Fine Utilization of Resources, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, Hunan, P. R. China.
Key Laboratory of Protein Chemistry and Developmental Biology of State Education Ministry of China, College of Life Sciences, Hunan Normal University, Changsha, Hunan, P. R. China.
PLoS One. 2017 Jul 3;12(7):e0180276. doi: 10.1371/journal.pone.0180276. eCollection 2017.
Glycolipids might become a new type of promising non-viral gene delivery systems because of their low cytotoxicity, structural diversity, controllable aqua- and lipo-solubility, appropriate density and distribution of positive charges, high transfer efficiency and potential targeting function. In this study, four kinds of L-arabinose-based cationic glycolipids (Ara-DiC12MA, Ara-DiC14MA, Ara-DiC16MA and Ara-DiC18MA) containing quaternary ammonium as hydrophilic headgroup and two alkane chains as hydrophobic domain were synthesized and characterized. They were observed to have strong affinities for plasmid DNA (pDNA) and siRNA, the pDNA can be completely condensed at N/P ratio less than 2, and the siRNA can be completely retarded at N/P ratio less than 3. The dynamic light scattering (DLS) experiment and atomic force microscopy (AFM) experiment demonstrated that cationic lipids and their lipoplexes possessed suitable particle sizes with near-spherical shape and proper ζ-potentials for cell transfection. The Ara-DiC16MA liposome was found to have good transfection efficacy in HEK293, PC-3 and Mat cells compared with other three kinds of liposomes, and also maintain low cytotoxicity and better uptake capability in vitro. Furthermore, the gene silencing assay showed that Ara-DiC14MA and Ara-DiC16MA liposomes have demonstrated effective delivery and higher gene knockdown activity (>80%) in the above mentioned cells than Lipofectamine 2000. These results indicated Ara-DiC16MA can be developed for efficient and low toxic gene delivery.
糖脂因其低细胞毒性、结构多样性、可控的水溶性和脂溶性、合适的正电荷密度和分布、高转移效率以及潜在的靶向功能，可能成为一种新型的有前景的非病毒基因递送系统。在本研究中，合成并表征了四种基于L-阿拉伯糖的阳离子糖脂（Ara-DiC12MA、Ara-DiC14MA、Ara-DiC16MA和Ara-DiC18MA），它们含有季铵作为亲水头部基团和两条烷烃链作为疏水结构域。观察到它们对质粒DNA（pDNA）和siRNA具有很强的亲和力，pDNA在N/P比小于2时可完全凝聚，siRNA在N/P比小于3时可完全阻滞。动态光散射（DLS）实验和原子力显微镜（AFM）实验表明，阳离子脂质及其脂质体具有适合细胞转染的粒径，呈近球形且具有合适的ζ电位。与其他三种脂质体相比，发现Ara-DiC16MA脂质体在HEK293、PC-3和Mat细胞中具有良好的转染效果，并且在体外保持低细胞毒性和更好的摄取能力。此外，基因沉默试验表明，Ara-DiC14MA和Ara-DiC16MA脂质体在上述细胞中比Lipofectamine 2000表现出有效的递送和更高的基因敲低活性（>80%）。这些结果表明Ara-DiC16MA可被开发用于高效低毒的基因递送。
