Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China.
College of Food Science, Northeast Agricultural University, Harbin 150030, China.
Molecules. 2019 Jan 2;24(1):148. doi: 10.3390/molecules24010148.
The intestinal epithelial barrier plays a key protective role in the gut lumen. Bovine lactoferrin (bLF) has been reported to improve the intestinal epithelial barrier function, but its impact on tight junction (TJ) proteins has been rarely described. Human intestinal epithelial crypt cells (HIECs) were more similar to those in the human small intestine, compared with the well-established Caco-2 cells. Accordingly, both HIECs and Caco-2 cells were investigated in this study to determine the effects of bioactive protein bLF on their growth promotion and intestinal barrier function. The results showed that bLF promoted cell growth and arrested cell-cycle progression at the G2/M-phase. Moreover, bLF decreased paracellular permeability and increased alkaline phosphatase activity and transepithelial electrical resistance, strengthening barrier function. Immunofluorescence, western blot and quantitative real-time polymerase chain reaction revealed that bLF significantly increased the expression of three tight junction proteins-claudin-1, occludin, and ZO-1-at both the mRNA and protein levels, and consequently strengthened the barrier function of the two cell models. bLF in general showed higher activity in Caco-2 cells, however, HIECs also exhibited desired responses to barrier function. Therefore, bLF may be incorporated into functional foods for treatment of inflammatory bowel diseases which are caused by loss of barrier integrity.
肠道上皮屏障在肠道腔中起着关键的保护作用。牛乳铁蛋白(bLF)已被报道能改善肠道上皮屏障功能,但它对紧密连接(TJ)蛋白的影响却很少被描述。与成熟的 Caco-2 细胞相比,人肠道上皮隐窝细胞(HIEC)与人小肠中的细胞更为相似。因此,本研究同时考察了 HIEC 和 Caco-2 细胞,以确定生物活性蛋白 bLF 对其生长促进和肠道屏障功能的影响。结果表明,bLF 促进细胞生长,并将细胞周期阻滞在 G2/M 期。此外,bLF 降低了细胞旁通透性,增加了碱性磷酸酶活性和跨上皮电阻,增强了屏障功能。免疫荧光、Western blot 和实时定量聚合酶链反应显示,bLF 显著增加了三种紧密连接蛋白- Claudin-1、Occludin 和 ZO-1 的表达,mRNA 和蛋白水平均有增加,从而增强了两种细胞模型的屏障功能。bLF 在 Caco-2 细胞中总体表现出更高的活性,但 HIEC 对屏障功能也表现出了期望的反应。因此,bLF 可能被纳入功能性食品中,用于治疗因屏障完整性丧失而引起的炎症性肠病。