Baltes Steffen, Nau Heinz, Lampen Alfonso
Institute for Food Toxicology, University of Veterinary Medicine Hannover, Foundation Bischofsholer Damm 15, 30173 Hannover, Germany.
Dev Growth Differ. 2004 Dec;46(6):503-14. doi: 10.1111/j.1440-169x.2004.00765.x.
Vitamin A and retinoids are essential nutrients for the differentiation of epithelia. Vitamin A deficiency is accompanied by an impairment in intestinal integrity. We investigated whether retinoids influence the differentiation and permeability of Caco-2 cells under serum-free culture conditions as a model for the intestinal epithelium. Treatment of the Caco-2 cells with retinoic acids (RA) resulted in an increased specific activity, enhanced mRNA expression, and induction of the 5'-flanking promoter activity of the marker enzyme for the differentiation intestinal alkaline phosphatase. Surprisingly, permeability of the Caco-2 monolayer, as measured by transepithelial electric resistance and [3H]-mannitol flux, was found to be enhanced by RA. Treatment with RA had only a slight effect on the mRNA expression of the tight junction-associated proteins occludin, ZO-1, claudin-1, -3, and -4, but enhanced the expression of claudin-2, which was recently suggested to form a paracellular ion channel. The role of retinoids as potent inducers of epithelial differentiation was confirmed for the Caco-2 cells under serum-free culture conditions and it was concluded that IAP is a target gene of RA. The inverse regulation of the permeability by RA under these serum-free conditions showed that other mechanisms, which are essential to regulate intestinal epithelial integrity with respect to decreased permeability, have to be identified.
维生素A和类视黄醇是上皮细胞分化所必需的营养素。维生素A缺乏会伴随着肠道完整性受损。我们研究了在无血清培养条件下,类视黄醇是否会影响作为肠上皮模型的Caco-2细胞的分化和通透性。用视黄酸(RA)处理Caco-2细胞会导致分化型肠碱性磷酸酶标记酶的比活性增加、mRNA表达增强以及5'-侧翼启动子活性的诱导。令人惊讶的是,通过跨上皮电阻和[3H]-甘露醇通量测量发现,RA可增强Caco-2单层细胞的通透性。用RA处理对紧密连接相关蛋白闭合蛋白、ZO-1、闭合蛋白-1、-3和-4的mRNA表达只有轻微影响,但增强了闭合蛋白-2的表达,最近有人提出闭合蛋白-2可形成细胞旁离子通道。在无血清培养条件下,类视黄醇作为上皮分化的有效诱导剂对Caco-2细胞的作用得到了证实,并且得出结论IAP是RA的靶基因。在这些无血清条件下,RA对通透性的反向调节表明,必须确定其他对于降低通透性以调节肠上皮完整性至关重要的机制。