Claudin-binder C-CPE mutants enhance permeability of insulin across human nasal epithelial cells
Claudin-binder C-CPE mutants enhance permeability of insulin across human nasal epithelial cells
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DOI：
10.3109/10717544.2015.1050530
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发表时间：
2016-10-01
期刊：
影响因子：
6
通讯作者：
Himi, Tetsuo
中科院分区：
文献类型：
--
作者：
Kojima, Takashi;Kondoh, Masuo;Himi, Tetsuo
目的：鼻内胰岛素给药具有治疗阿尔茨海默病的潜力，并且在鼻内通过鼻粘膜给药时，由紧密连接调节的细胞旁途径是重要的。产气荚膜梭菌肠毒素（C-CPE）的C末端片段结合紧密连接蛋白claudin并破坏紧密连接屏障而没有细胞毒性作用。被称为C-CPE 194的C-CPE突变体仅与claudin-4结合，而被称为C-CPE m19的C-CPE 194突变体不仅与claudin-4结合，还与claudin-1结合。为了研究C-CPE突变体对人鼻上皮细胞紧密连接功能和重组人胰岛素跨细胞渗透性的影响，结果：C-CPE 194和C-CPE m19可破坏HNECs的屏障和栅栏功能，但不影响claudin-1、claudin-4、claudin-7的表达，也不影响细胞毒性，但可短暂增加ERK 1/2磷酸化活性。用MAPKK抑制剂U 0126预处理可以防止C-CPE 194和C-CPE m19引起的屏障功能破坏。C-CPE 194和C-CPE m19能显著增强重组人胰岛素对HNECs的渗透性，U 0126也能抑制其渗透性。这些发现表明，C-CPE突变体m194和m19可以通过MAPK途径调节胰岛素对HNECs的通透性，并可能通过直接介导胰岛素对HNECs通透性的调节，在阿尔茨海默病等疾病的治疗中发挥重要作用。鼻内胰岛素给药。
Objective: Intranasal insulin administration has therapeutic potential for Alzheimer's disease and in intranasal administration across the nasal mucosa, the paracellular pathway regulated by tight junctions is important. The C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE) binds the tight junction protein claudin and disrupts the tight junctional barrier without a cytotoxic effect. The C-CPE mutant called C-CPE 194 binds only to claudin-4, whereas the C-CPE 194 mutant called C-CPE m19 binds not only to claudin-4 but also to claudin-1.Methods: In the present study, to investigate the effects of C-CPE mutants on the tight junctional functions of human nasal epithelial cells (HNECs) and on the permeability of human recombinant insulin across the cells, HNECs were treated with C-CPE 194 and C-CPE m19.Results: C-CPE 194 and C-CPE m19 disrupted the barrier and fence functions without changes in expression of claudin-1, -4, -7, and occludin or cytotoxicity, whereas they transiently increased the activity of ERK1/2 phosphorylation. The disruption of the barrier function caused by C-CPE 194 and C-CPE m19 was prevented by pretreatment with the MAPKK inhibitor U0126. Furthermore, C-CPE 194 and C-CPE m19 significantly enhanced the permeability of human recombinant insulin across HNECs and the permeability was also inhibited by U0126.Conclusion: These findings suggest that C-CPE mutants 194 and m19 can regulate the permeability of insulin across HNECs via the MAPK pathway and may play a crucial role in therapy for the diseases such as Alzheimer's disease via the direct intranasal insulin administration.