Vishnevskaia O N, Burdin D V, Gorshkov A N, Grefner N M, Markov A G
Ross Fiziol Zh Im I M Sechenova. 2013 Jan;99(1):81-91.
Action of polycation protein protamine on the expression of tight junction proteins (claudins-1, -2, -3 and occludin) which contribute to paracellular transport function was investigated on cellular models of tight (MDCK I cell line) and leaky (Caco-2 cell line) epithelium. The expression of claudins-1,-3 and occludin was observed in both cell lines by methods of immunocytochemistry. Influence of protamine (100 microg/ml; 30 min; apical) on fluorescence intensity of claudins-1, -3 was different in MDCK I and Caco-2 cells. Addition ofprotamine to the incubation medium of Caco-2 cells resulted in significant increase of claudin-3 expression by 45 % (p <0.01) in comparison with control, whereas claudin-1 and occludin expression did not alter. On the contrary, in MDCK I cells protamine induced the significant decrease ofclaudin-1 and -3 expression by 25 % (p <0.001) and 15 % (p < 0.01) respectively, whereas occludin expression did not alter. It was confirmed by the methods of confocal laser scanning microscopy that protamine alter the expression of claudins-1, -3 directly in the tight junctions. Our results suggest that charged chyme components may alter paracellular permeability of epithelium.
在紧密（MDCK I细胞系）和渗漏（Caco-2细胞系）上皮细胞模型上，研究了聚阳离子蛋白鱼精蛋白对有助于细胞旁转运功能的紧密连接蛋白（闭合蛋白-1、-2、-3和封闭蛋白）表达的作用。通过免疫细胞化学方法在两种细胞系中均观察到了闭合蛋白-1、-3和封闭蛋白的表达。鱼精蛋白（100微克/毫升；30分钟；顶端）对MDCK I和Caco-2细胞中闭合蛋白-1、-3荧光强度的影响不同。与对照相比，向Caco-2细胞的孵育培养基中添加鱼精蛋白导致闭合蛋白-3表达显著增加45%（p<0.01），而闭合蛋白-1和封闭蛋白的表达未改变。相反，在MDCK I细胞中，鱼精蛋白分别导致闭合蛋白-1和-3表达显著降低25%（p<0.001）和15%（p<0.01），而封闭蛋白的表达未改变。共聚焦激光扫描显微镜方法证实，鱼精蛋白直接改变紧密连接处闭合蛋白-1、-3的表达。我们的结果表明，带电荷的食糜成分可能会改变上皮细胞的细胞旁通透性。
