Effect of chitosan on epithelial permeability and structure
Effect of chitosan on epithelial permeability and structure
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DOI：
10.1016/s0378-5173(99)00030-7
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发表时间：
1999-05-10
影响因子：
5.8
通讯作者：
Merwin, JR
中科院分区：
文献类型：
--
作者：
Dodane, V;Khan, MA;Merwin, JR
大量研究表明，壳聚糖作为一种粘膜粘附聚合物，是一种潜在的透粘膜给药促进剂。为了进一步了解壳聚糖对粘膜屏障作用的机制，研究了壳聚糖对肠上皮细胞单层功能和结构的活性。在Caco-2细胞，壳聚糖引起可逆的，时间和剂量依赖性的跨上皮电阻下降。壳聚糖对紧密连接的影响证实了增加的渗透系数甘露醇运输时，细胞用0.1 - 0.5%w/v壳聚糖溶液处理60分钟相比，对照细胞。紧密连接的参与可视化共聚焦扫描显微镜使用occludin和ZO-1，紧密连接蛋白。孵育后，与0.01或0.1%w/v壳聚糖，标记的两种蛋白质的不同定位和荧光强度降低在细胞周边。此外，在细胞与细胞接触的区域优先观察到肌动蛋白的局灶性凝聚。然而，在24小时恢复后，细胞结构类似于未处理的对照细胞。同时添加放线菌酮，蛋白质合成抑制剂，防止完全恢复。这意味着蛋白质合成是细胞恢复到基线水平所必需的。壳聚糖治疗似乎略有扰动的质膜评估增加释放的乳酸脱氢酶。然而，如台盼蓝染料排除所示，添加0.5%壳聚糖60 mill不影响细胞活力。这些数据表明，壳聚糖通过影响上皮细胞的细胞旁和细胞内途径，以可逆的方式增加细胞通透性。(C)1999年由Elsevier Science B.V.出版，版权所有。
Numerous studies have shown that chitosan, a mucoadhesive polymer, is a potential enhancer for transmucosal drug delivery. To further understand the mechanisms involved in chitosan action on the mucosal barrier, the activity of chitosan on the function and structure of monolayers of intestinal epithelial cells was investigated. In Caco-2 cells, chitosan caused a reversible, time and dose-dependent decrease in transepithelial electrical resistance. The effect of chitosan on tight junctions was confirmed by an increased permeability coefficient for mannitol transport when cells were treated with 0.1-0.5% w/v chitosan solution for 60 min compared to control cells. Involvement of tight junctions was visualized by confocal scanning microscopy using occludin and ZO-1, tight junctional proteins. Following an incubation with 0.01 or 0.1% w/v chitosan, labeling of both proteins varied in localization and decreased in fluorescent intensity at the cell periphery. In addition, a focal condensation of actin was observed preferentially at areas of cell-to-cell contacts. However, after 24-h recovery, the cell structure resembled untreated control cells. Simultaneous addition of cycloheximide, a protein synthesis inhibitor, prevented full recovery. This implied that protein synthesis was required for the cells to return to baseline levels. Chitosan treatment appeared to slightly perturb the plasma membrane as assessed by an increased release of lactate dehydrogenase. However, addition of 0.5% chitosan for 60 mill did not affect cell viability as shown by Trypan blue dye exclusion. These data suggest that chitosan increases cell permeability by affecting paracellular and intracellular pathways of epithelial cells, in a reversible manner. (C) 1999 Published by Elsevier Science B.V. All rights reserved.