Abstract
High mobility group box 1 protein (HMGB1) is involved in the pathogenesis of inflammatory bowel disease (IBD). Patients with IBD develop zinc deficiency. However, the detailed roles of HMGB1 and zinc deficiency in the intestinal epithelial barrier and cellular metabolism of IBD remain unknown. In the present study, Caco-2 cells in 2D culture and 2.5D Matrigel culture were pretreated with transforming growth factor-β (TGF-β) type 1 receptor kinase inhibitor EW-7197, epidermal growth factor receptor (EGFR) kinase inhibitor AG-1478 and a TNFα antibody before treatment with HMGB1 and inflammatory cytokines (TNFα and IFNγ). EW-7197, AG-1478 and the TNFα antibody prevented hyperpermeability induced by HMGB1 and inflammatory cytokines in 2.5D culture. HMGB1 affected cilia formation in 2.5D culture. EW-7197, AG-1478 and the TNFα antibody prevented the increase in cell metabolism induced by HMGB1 and inflammatory cytokines in 2D culture. Furthermore, ZnSO4 prevented the hyperpermeability induced by zinc chelator TPEN in 2.5D culture. ZnSO4 and TPEN induced cellular metabolism in 2D culture. The disruption of the epithelial barrier induced by HMGB1 and inflammatory cytokines contributed to TGF-β/EGF signaling in Caco-2 cells. The TNFα antibody and ZnSO4 as well as EW-7197 and AG-1478 may have potential for use in therapy for IBD.
Highlights
High mobility group box 1 (HMGB1) is a chromatin-associated protein and one of the damage-associated molecular patterns (DAMPs) [1]
We have previously reported that HMGB1-downregulated angulin-1/LSR induces epithelial barrier disruption via the tight junction (TJ) molecule claudin-2 (CLDN-2) in Calu-3 cells [11]
We have reported that HMGB1 enhances epithelial permeability via p63/transforming growth factor-β (TGF-β) signaling in lung and terminal bronchial epithelial cells [12]
Summary
High mobility group box 1 (HMGB1) is a chromatin-associated protein and one of the damage-associated molecular patterns (DAMPs) [1]. Many recent studies have indicated that HMGB1 protein is involved in the development of inflammatory bowel disease (IBD) [4]. AG-1478 is an epidermal growth factor receptor (EGFR) kinase inhibitor that, like the MAP kinase inhibitor U0126, prevents the disassembly of tight junction (TJ) transmembrane proteins caused by inflammatory cytokines such as, TNFα, IL-4 and IFNγ in Calu-3 airway epithelial cells [10]. We have previously reported that HMGB1-downregulated angulin-1/LSR induces epithelial barrier disruption via the TJ molecule claudin-2 (CLDN-2) in Calu-3 cells [11]. Many studies have indicated that TJ molecules play important roles in the intestinal epithelial barrier of IBD [19]. The detailed mechanisms by which HMGB1 and zinc affect TJ molecules via tTJ in the intestinal epithelial barrier of IBD have not been elucidated. We analyzed Caco-2 monolayer cells (2D culture) and Caco-2 spheroid cells (2.5D Matrigel culture), which have lumen formation, with the aim of developing a new therapy for intestinal inflammation and injury in patients with IBD
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