Abstract
Mannan oligosaccharide (MOS) is well-known as an effective fed supplement for livestock to increase their nutrients absorption and health status. Pentasaccharide of mannan (MOS5) was reported as a molecule that possesses the ability to increase tight junction of epithelial tissue, but the structure and mechanism of action remains undetermined. In this study, the mechanism of action and structure of MOS5 were investigated. T84 cells were cultured and treated with MOS5 compared with vehicle and compound C, a 5′-adenosine monophosphate-activated protein kinase (AMPK) inhibitor. The results demonstrated that the ability of MOS5 to increase tight junction integration was inhibited in the presence of dorsomorphine (compound C). Phosphorylation level of AMPK was elevated in MOS5 treated group as determined by Western blot analysis. Determination of MOS5 structure was performed using enzymatic mapping together with 1H, 13C NMR, and 2D-NMR analysis. The results demonstrated that the structure of MOS5 is a β-(1,4)-mannotetraose with α-(1,6)-galactose attached at the second mannose unit from non-reducing end.
Highlights
Tight junction is one of the crucial compartments of a barrier function of an epithelial tissue which is a part of anatomical barriers of innate immunity in complex and higher organisms.An impaired tight junction is a common pathologic feature in many inflammatory diseases such as inflammatory bowel disease (IBD) or Crohn’s disease or even chronic diarrhea in HIV-infected patient, or a side-effect of some drugs such as gefitinib or other drugs in EFGR-inhibitor family [1,2,3]
MOS5, a pentasaccharide obtained from the digestion of pretreated galactomannan from copra meal with recombinant β-mannanase was reported to have the ability to enhance tight junction integration in epithelial cells [24]
Previous reports have demonstrated that β-glycan enhances tight junction integration of epithelial cells through activation of the AMPK pathway [12,13,25,26]
Summary
Tight junction is one of the crucial compartments of a barrier function of an epithelial tissue which is a part of anatomical barriers of innate immunity in complex and higher organisms. Many recent studies had reported that β-glycans can increase tight junction formation. MOS5, a pentasaccharide obtained from the digestion of pretreated galactomannan from copra meal with recombinant β-mannanase was reported to have the ability to enhance tight junction integration in epithelial cells [24]. Previous reports have demonstrated that β-glycan enhances tight junction integration of epithelial cells through activation of the AMPK pathway [12,13,25,26]. Dorsomorphine (compound C), an inhibitor of AMPK, was used to elucidate the involvement of AMPK in the activation of tight junction integration of epithelial cells by MOS5. MOS5 structure was successfully elucidated using specific enzymatic mapping and 1 H, 13 C NMR, and 2D NMR analysis
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