Role of regenerating gene I in claudin expression and barrier function in the small intestine
Role of regenerating gene I in claudin expression and barrier function in the small intestine
复制标题
DOI：
10.1016/j.trsl.2016.03.007
复制
发表时间：
2016-07-01
影响因子：
7.8
通讯作者：
Miwa, Hiroto
中科院分区：
文献类型：
--
作者：
Kitayama, Yoshitaka;Fukui, Hirokazu;Miwa, Hiroto
我们最近发现，再生基因(REG)I的缺失会导致非类固醇抗炎药引起的胃肠道损伤。然而，Reg I对这种病理生理条件起保护作用的机制尚不清楚。在这里，我们研究了Reg I是否在小肠紧密连接蛋白的诱导和粘膜屏障功能中发挥作用。用FITC-葡聚糖法和跨皮细胞电阻(TEER)法检测Reg I基因缺陷小鼠的小肠通透性。用体外培养的Caco 2细胞检测Reg Iα对TEER、Claudins表达和细胞内信号转导的影响。在活体条件下，研究了claudins 3和4在小鼠小肠中的表达。Reg I缺乏可显著降低小肠上皮细胞claudin 3的表达。经消炎痛处理后，REG-1基因敲除小鼠血清FITC-葡聚糖水平显著高于野生型(WT)小鼠。正常情况下，与WT小鼠相比，REG Iα基因敲除小鼠的小肠TEER水平显著降低。REG Iα刺激可显著提高Caco 2细胞的TEER水平。Reg 1a可增强Caco2细胞中Claudins 3和4的表达，并促进Sp1、Akt和ERK的磷酸化，而这些作用可被抗Reg Iα抗体减弱。Reg I可能通过诱导紧密连接蛋白，如Claudins 3和4，在维持粘膜屏障功能中发挥作用。
We have recently shown that loss of the regenerating gene (Reg) I causes susceptibility to nonsteroidal anti-inflammatory drug-induced gastrointestinal damage. However, the mechanism by which Reg I plays a protective role against this pathophysiological condition is unclear. Here, we investigated whether Reg I plays roles in the induction of tight junction proteins and mucosal barrier function in the small intestine. The small-intestinal permeability was evaluated in Reg I-deficient mice by FITC-dextran and transepithelial electrical resistance (TEER) assay. The effect of REG I alpha on TEER, claudins expression, and intracellular signaling was examined using Caco2 cells in vitro. Small-intestinal expression of claudins 3 and 4 was investigated in Reg I-deficient mice in vivo. REG I deficiency significantly decreased the expression of claudin 3 in the small-intestinal epithelium. When mice were treated with indomethacin, the serum level of FITC-dextran in Reg 1 knockout mice was significantly higher than that in wild-type (WT) mice. The level of small-intestinal TEER was significantly decreased in Reg I alpha knockout mice compared with WT mice under normal condition. REG I alpha stimulation significantly enhanced the level of TEER in Caco2 cells. Treatment with REG la enhanced the expression of claudins 3 and 4 and promoted Sp1, Akt, and ERK phosphorylation in Caco2 cells, whereas these effects were attenuated by treatment with anti-REG I alpha antibody. Reg I may play a role in the maintenance of mucosal barrier function by inducing tight junction proteins such as claudins 3 and 4.