Graduate School of Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki, 305-8572, Japan.
Cytotechnology. 2005 Jan;47(1-3):89-96. doi: 10.1007/s10616-005-3756-5.
On the basis of transepithelial electrical resistance (TER) measurements, we found that capsaicin (100 muM)-treated human intestinal Caco-2 cells show a momentary increase in tight-junction (TJ) permeability (decrease in TER) followed by a complete recovery. We used proteome analysis to search for proteins that are associated with the recovery of TJ permeability in capsaicin-treated Caco-2 cells. A protein with a relative molecular mass of 14 kDa was found to be expressed more highly in capsaicin-treated cells than in nontreated cells. Mass spectrometry and sequence analyses revealed that the protein that is expressed significantly upon capsaicin treatment is the ribosomal protein P2; its cDNA sequence was identical to that found in the human genome database. An increase in the amount of cellular filamentous actin (F-actin) was shown after 8 h of incubation with capsaicin. It has been reported that P2 activates elongation factor 2, which stabilizes F-actin filaments, and that the depolymerization of F-actin is associated with the increase in TJ permeability (decrease in TER). Consequently, these results suggest that P2 plays an important role in the recovery of the TJ permeability in capsaicin-treated human intestinal cells.
基于跨上皮电阻 (TER) 测量,我们发现辣椒素 (100μM) 处理的人肠道 Caco-2 细胞表现出紧密连接 (TJ) 通透性的瞬间增加 (TER 降低),随后完全恢复。我们使用蛋白质组分析来寻找与辣椒素处理的 Caco-2 细胞 TJ 通透性恢复相关的蛋白质。发现一种相对分子质量为 14 kDa 的蛋白质在辣椒素处理的细胞中表达水平高于未处理的细胞。质谱和序列分析表明,在辣椒素处理后显著表达的蛋白质是核糖体蛋白 P2;其 cDNA 序列与人基因组数据库中发现的完全相同。在用辣椒素孵育 8 小时后,细胞丝状肌动蛋白 (F-actin) 的量增加。据报道,P2 激活延伸因子 2,稳定 F-actin 纤维,而 F-actin 的解聚与 TJ 通透性的增加 (TER 降低) 有关。因此,这些结果表明 P2 在辣椒素处理的人肠道细胞 TJ 通透性恢复中发挥重要作用。