Freeman Jennifer J, Feng Yongjia, Demehri Farokh R, Dempsey Peter J, Teitelbaum Daniel H
*Department of Surgery, Section of Pediatric Surgery, and Center for Organogenesis, University of Michigan, Ann Arbor, Michigan, USA; and School of Medicine, Department of Pediatrics, University of Colorado, Denver, Anschutz Medical Campus, Denver, Colorado, USA.
*Department of Surgery, Section of Pediatric Surgery, and Center for Organogenesis, University of Michigan, Ann Arbor, Michigan, USA; and School of Medicine, Department of Pediatrics, University of Colorado, Denver, Anschutz Medical Campus, Denver, Colorado, USA
FASEB J. 2015 Jul;29(7):2943-58. doi: 10.1096/fj.14-269480. Epub 2015 Mar 17.
Recent studies suggest a close interaction between epidermal growth factor (EGF) and TLR signaling in the modulation of intestinal epithelial cell (IEC) proliferation; however, how these signaling pathways adjust IEC proliferation is poorly understood. We utilized a model of total parenteral nutrition (TPN), or enteral nutrient deprivation, to study this interaction as TPN results in mucosal atrophy due to decreased IEC proliferation and increased apoptosis. We identified the novel finding of decreased mucosal atrophy in TLR4 knockout (TLR4KO) mice receiving TPN. We hypothesized that EGF signaling is preserved in TLR4KO-TPN mice and prevents mucosal atrophy. C57Bl/6 and strain-matched TLR4KO mice were provided either enteral feeding or TPN. IEC proliferation and apoptosis were measured. Cytokine and growth factor abundances were detected in both groups. To examine interdependence of these pathways, ErbB1 pharmacologic blockade was used. The marked decline in IEC proliferation with TPN was nearly prevented in TLR4KO mice, and intestinal length was partially preserved. EGF was significantly increased, and TNF-α decreased in TLR4KO-TPN versus wild-type (WT)-TPN mice. Apoptotic positive crypt cells were 15-fold higher in WT-TPN versus TLR4KO-TPN mice. Bcl-2 was significantly increased in TLR4KO-TPN mice, while Bax decreased 10-fold. ErbB1 blockade prevented this otherwise protective effect in TLR4KO-sTPN mice. TLR4 blockade significantly prevented TPN-associated atrophy by preserving proliferation and preventing apoptosis. This is driven by a reduction in TNF-α abundance and increased EGF. Potential manipulation of this regulatory pathway may have significant clinical potential to prevent TPN-associated atrophy.
近期研究表明,表皮生长因子(EGF)与Toll样受体(TLR)信号通路在调节肠道上皮细胞(IEC)增殖过程中存在密切相互作用;然而,这些信号通路如何调节IEC增殖却鲜为人知。我们利用全肠外营养(TPN)模型,即肠内营养剥夺模型,来研究这种相互作用,因为TPN会导致IEC增殖减少和细胞凋亡增加,进而引起黏膜萎缩。我们发现了一个新现象,即接受TPN的Toll样受体4基因敲除(TLR4KO)小鼠的黏膜萎缩程度减轻。我们推测,在TLR4KO-TPN小鼠中,EGF信号通路得以保留,从而防止了黏膜萎缩。给C57Bl/6小鼠和品系匹配的TLR4KO小鼠提供肠内喂养或TPN。测量IEC的增殖和凋亡情况。检测两组细胞因子和生长因子的丰度。为了研究这些信号通路的相互依赖性,使用了ErbB1药物阻断剂。TLR4KO小鼠中,TPN导致的IEC增殖显著下降几乎得到了预防,肠道长度也得到了部分保留。与野生型(WT)-TPN小鼠相比,TLR4KO-TPN小鼠中EGF显著增加,而肿瘤坏死因子-α(TNF-α)减少。WT-TPN小鼠中凋亡阳性隐窝细胞比TLR4KO-TPN小鼠高15倍。TLR4KO-TPN小鼠中Bcl-2显著增加,而Bax减少了10倍。ErbB1阻断剂消除了TLR4KO-TPN小鼠原本具有的这种保护作用。TLR4阻断通过维持增殖和防止凋亡,显著预防了TPN相关的萎缩。这是由TNF-α丰度降低和EGF增加所驱动的。对这一调节通路的潜在调控可能具有预防TPN相关萎缩的重大临床潜力。