Protective role of 1,25(OH)2 vitamin D3 in the mucosal injury and epithelial barrier disruption in DSS-induced acute colitis in mice.
BACKGROUND
Intestinal hyper-permeability plays a critical role in the etiopathogenesis of inflammatory bowel disease (IBD) by affecting the penetration of pathogens, toxic compounds and macromolecules. 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], the active form of vitamin D, has been shown to be an important regulator of IBD and recent epidemiology suggests that patients with IBD have an impaired vitamin D status. The purpose of this study is to investigate the possible protective effects of 1,25(OH)2D3 on mucosal injury and epithelial barrier disruption on dextran sulfate sodium (DSS)-induced acute colitis model.


METHODS
We used DSS-induced acute colitis model to investigate the protective effects of 1,25(OH)2D3 on mucosal injury and epithelial barrier integrity. Severity of colitis was evaluated by disease activity index (DAI), body weight (BW) change, colon length, histology, myeloperoxidase (MPO) activity, and proinflammatory cytokine production including tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). In vitro the protective role of 1,25(OH)2D3 was assessed by incubating Caco-2 cells with or without DSS and measuring transepithelial electrical resistance (TEER) and fluorescein isothiocyanate dextran (FITC-D). The intestinal permeability was analyzed by FITC-D, bacterial translocation and measurement of lipopolysaccharide (LPS). Ultrastructural features of the colon tissue and Caco-2 cell monolayer were observed by electron microscopy. Expressions of tight junction (TJ) proteins in the colon mucosa and Caco-2 cells were detected by immunohistochemistry, immunofluorescence, Western blot and real-time fluorescent quantitative PCR, respectively.


RESULTS
DSS-induced acute colitis model was characterized by a reduced BW, AUC of BW, serum calcium, higher DAI, AUC of DAI, shortened colon length, elevated MPO activity, worsened histologic inflammation, increased mononuclear cell numbers in mesenteric lymph nodes (MLNs) and colonic lamina propria (LP), and enhanced proteins and mRNA levels of TNF-α and IFN-γ. 1,25(OH)2D3 markedly increased expressions of TJ proteins and mRNA and decreased the FITC-D permeability and the level of LPS. Furthermore, 1,25(OH)2D3 abrogated bacterial translocation to MLNs and ameliorated ultrastructural features of the colon epithelium by scanning electron microscopy (SEM). In vitro, 1,25(OH)2D3 increased TEER, TJ proteins and mRNA expressions, decreased the FITC-D permeability, and preserved structural integrity of the TJ in Caco-2 cells.


CONCLUSIONS
1,25(OH)2D3 may play a protective role in mucosal barrier homeostasis by maintaining the integrity of junction complexes and in healing capacity of the colon epithelium. 1,25(OH)2D3 may represent an attractive and novel therapeutic agent for the adjuvant therapy of IBD.
Department of Gastroenterology, The Second Hospital of Hebei Medical University, Hebei Key Laboratory of Gastroenterology, Hebei Institute of Gastroenterology, No,215 Heping West Road, 050000, Shijiazhuang, China.
Hongwei
H
Zhao
Hong
H
Zhang
Hui
H
Wu
Hui
H
Li
Lei
L
Liu
Jian
J
Guo
Chenyang
C
Li
David Q
DQ
Shih
Xiaolan
X
Zhang
D000208:Acute Disease; D000818:Animals; D018938:Caco-2 Cells; D002117:Calcitriol / Q000494:pharmacology / Q000627:therapeutic use*; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology*; D003092:Colitis / Q000139:chemically induced* / Q000503:physiopathology / Q000517:prevention & control*; D003110:Colonic Neoplasms / Q000473:pathology / Q000503:physiopathology; D016264:Dextran Sulfate / Q000009:adverse effects* / Q000494:pharmacology; D004195:Disease Models, Animal; D004848:Epithelium / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology; D006706:Homeostasis / Q000187:drug effects / Q000502:physiology; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects / Q000502:physiology; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019108:Tight Junctions / Q000187:drug effects / Q000648:ultrastructure
D016264:Dextran Sulfate; D002117:Calcitriol