Department of Cellular and Molecular Medicine, The Panum Institute, Faculty of Health Sciences, University of Copenhagen, Denmark.
Department of Cellular and Molecular Medicine, The Panum Institute, Faculty of Health Sciences, University of Copenhagen, Denmark.
Biochim Biophys Acta Biomembr. 2017 Mar;1859(3):360-367. doi: 10.1016/j.bbamem.2016.12.017. Epub 2016 Dec 27.
Chitosan is a polycationic polysaccharide consisting of β-(1-4)-linked glucosamine units and due to its mucoadhesive properties, chemical derivatives of chitosan are potential candidates as enhancers for transmucosal drug delivery. Recently, glycol chitosan (GC), a soluble derivative of chitosan, was shown to bind specifically to lipid raft domains in model bilayers. The small intestinal brush border membrane has a unique lipid raft composition with high amounts of glycolipids cross-linked by lectins, and the aim of the present work therefore was to study the interaction of FITC-conjugated GC (FITC-GC) with the small intestinal epithelium. Using organ culture of pig jejunal mucosal explants as a model system, we observed widespread binding of luminal FITC-GC to the brush border. Only little uptake via constitutive endocytosis into apical early endosomes occurred, unless endocytosis was induced by the simultaneous presence of cholera toxin B subunit (CTB). Biochemically, GC bound to microvillus membrane vesicles and caused a change in the density profile of detergent resistant membranes (DRMs). Collectively, the results showed that FITC-GC binds passively to lipid raft domains in the brush border, i.e. without inducing endocytosis like CTB. Instead, and unlike CTB, FITC-GC seems to exert a stabilizing, detergent-protective effect on the lipid raft organization of the brush border.
壳聚糖是一种带正电荷的多糖,由β-(1-4)-连接的葡萄糖胺单元组成。由于其具有黏膜黏附特性,壳聚糖的化学衍生物是作为跨黏膜药物传递增强剂的潜在候选物。最近,壳聚糖的可溶性衍生物——乙二醇壳聚糖(GC)被证明可以特异性结合模型双层膜中的脂筏域。小肠刷状缘膜具有独特的脂筏组成,其中含有大量通过凝集素交联的糖脂,因此本工作的目的是研究 FITC 标记的 GC(FITC-GC)与小肠上皮的相互作用。我们使用猪空肠黏膜外植体的器官培养作为模型系统,观察到腔内 FITC-GC 广泛结合到刷状缘。只有在霍乱毒素 B 亚单位(CTB)同时存在的情况下,通过组成型内吞作用进入顶端早期内体,才会发生少量摄取。从生物化学上讲,GC 结合到微绒毛膜囊泡上,并导致去污剂抗性膜(DRM)密度分布发生变化。总之,这些结果表明,FITC-GC 被动结合到刷状缘的脂筏域,即不像 CTB 那样诱导内吞作用。相反,与 CTB 不同,FITC-GC 似乎对刷状缘的脂筏组织发挥稳定、去污剂保护作用。