Human Milk Oligosaccharides Improve Tight Junction Proteins in Disrupted Intestinal Epithelial Cells
Keywords:Human milk, Human milk oligosaccharides, Tight junction protein, Disruption
The strength of intestinal barrier through tight junction proteins, such as Zonula Occludens-1 (ZO-1) and Occludin, is an important factor for nutrient digestion and absorption to promote growth of the body. Human milk oligosaccharides (HMOs) can protect the integrity of tight junction on the inflammation of intestinal epithelial cells. However, the distinct biological activity of HMOs in human milk on intestinal barrier and inflammation response remains unclear. This study aimed to investigate the patterns of HMOs in human milk and their preventive effects to the tight junction proteins disrupted by tumor necrosis factor-alpha (TNF-a). Breast milk samples from 23 Thai lactating women were separated using 3kDa cut-off column and then purified using solid phase extraction cartridges. The mass spectra of purified HMOs from individual milk samples were created using MALDI-TOF mass spectrometry. The effect of each pattern of HMOs on TNF-a-induced Caco-2 cell monolayer disruption was examined using transepithelial electrical resistance (TEER) measurement. The expression of ZO-1 and Occludin was assessed using immunofluorescence. Nine patterns of HMOs were found. The two major HMOs were galacto-oligosaccharides and sialyllactose (SL) group, particularly 3’SL and 6’SL. The level of TEER increased in the presence of HMOs, especially 3’SL and 6’SL. ZO-1 and Occludin were increasingly expressed in the presence of HMOs. In conclusion, this study suggests that HMO patterns of SL groups show the most protective effect on disruption of tight junction proteins.
Gabel G, Aschenbach JR, Muller F. Transfer of energy substrates across the ruminal epithelium: implications and limitations. Anim Health Res Rev 2002;3(4):15-30.
Furness JB, Rivera LR, Cho HJ, Bravo DM, Callaghan B. The gut as a sensory organ. Nat Rev Gastroenterol Hepatol 2013;10(12):729-40.
Chelakkot C, Ghim J, Ryu SH. Mechanisms regulating intestinal barrier integrity and its pathological implications. Exp Mol Med 2018;50(8):103.
Van der Flier LG, Clevers H. Stem cells, self-renewal, and differentiation in the intestinal epithelium. Annu Rev Physiol 2009;71:241-60.
Noah TK, Donahue B, Shroyer NF. Intestinal development and differentiation. Exp Cell Res 2011;317(19):2702-10.
Crosnier C, Stamataki D, Lewis J. Organizing cell renewal in the intestine: stem cells, signals and combinatorial control. Nat Rev Genet 2006;7(5):349-59.
Anderson RC, Gopal PK, Bassett S, Ellis A, Roy NC. The Role of Intestinal Barrier Function in Early Life in the Development of Colitis. InTech. 2012;1:1-30
Peterson LW, Artis D. Intestinal epithelial cells: regulators of barrier function and immune homeostasis. Nat Rev Immunol 2014;14(4):141-53.
Farkas AE, Capaldo CT, Nusrat A. Regulation of epithelial proliferation by tight junction proteins. Ann N Y Acad Sci 2012;1258:115-24.
Matter K, Aijaz S, Tsapara A, Balda MS. Mammalian tight junctions in the regulation of epithelial differentiation and proliferation. Curr Opin Cell Biol 2005;17(5):453-8.
Amasheh M, Fromm A, Krug SM, Amasheh S, Andres S, Zeitz M. Fromm M, Schulzke JD. TNFalpha-induced and berberine-antagonized tight junction barrierimpairment via tyrosine kinase, Akt and NFkappaB signaling. J Cell Sci 2010;123(Pt 23):4145-55.
Gitter AH, Bendfeldt K, Schulzke JD, Fromm M. Leaks in the epithelial barrier caused by spontaneous and TNF-alpha-induced single-cell apoptosis. FASEB J 2000;14(12):1749-53.
Kalliolias GD, Ivashkiv LB. TNF biology, pathogenic mechanisms and emerging therapeutic strategies. Nat Rev Rheumatol 2016;12(4):49-62.
Aggarwal BB. Signalling pathways of the TNF superfamily: a double-edged sword. Nat Rev Immunol 2003;3(9):745-56.
German JB, Freeman SL, Lebrilla CB, Mills DA. Human milk oligosaccharides: evolution, structures and bioselectivity as substrates for intestinal bacteria. Nestle Nutr Workshop Ser Pediatr Program 2008;62:205-18; discussion 18-22.
Zehra S, Khambati I, Vierhout M, Mian MF, Buck R, Forsythe P. Human Milk Oligosaccharides Attenuate Antigen-Antibody Complex Induced Chemokine Release from Human Intestinal Epithelial Cell Lines. J FoodSci 2018;83(2):499-508.
Azagra-Boronat I, Massot-Cladera M, Knipping K, Van't Land B, Stahl B, Garssen J, et al. Supplementation With 2'-FL and scGOS/lcFOS Ameliorates Rotavirus-Induced Diarrhea in Suckling Rats. Front Cell Infect Microbiol 2018;8:372.
Cai Y, Van Putten J P.M., Gilbert MS, Gerrits WJJ, Folkerts G, Braber S. Galacto-oligosaccharides as an anti-bacterial and anti-invasive agent in lung infections. Biomaterials 2022;283:121461.
Susana Figueroa-Lozano RA, Martin Beukema, Sander S.van Leeuwen, Lubbert Dijkhuizen, Paulde Vos. 2′-Fucosyllactose impacts the expression of mucus-related genes in goblet cells and maintains barrier function of gut epithelial cells. J Func Foods 2021;85.
Natividad JM, Rytz A, Keddani S, Bergonzelli G, Garcia-Rodenas CL. Blends of Human Milk Oligosaccharides Confer Intestinal Epithelial Barrier Protection in Vitro. Nutrients 2020;12(10).
Phoonlapdacha P, Tangshewinsirikul C, Phosuwattanakul J, Kittisakmontri K, et al. Gut microbiome profiles in Thai healthy pregnant woman and its association with types of foods. BMC Pregnancy Childbirth 2022; 22(1):79.
Chiangjong W, Bhakdi SC, Woramongkolchai N, Vanichapol T, Pongsakul N, Hongeng S, Chutipongtanate S. Cell-main spectra profile screening technique in simulation of circulating tumour cells using MALDI-TOF mass spectrometry. Cancer (Basal) 2021;13(15):3775.
Akbari P, Braber S, Alizadeh A, Verheijden KA, Schoterman MH, Kraneveld AD, et al. Galacto-oligosaccharides Protect the Intestinal Barrier by Maintaining the Tight Junction Network and Modulating the Inflammatory responses after a Challenge with the mycotoxin Deoxynivalenol in Human Caco-2 Cell Monolayers and B6C3F1 Mice. J Nutr 2015;145(7):1604-13.
Donovan SM, Comstock SS. Human milk oligosaccharides influence neonatal mucosal and systemic immunity. Ann Nutr Metab 2016;69(2):42-51.
Mehta A, Silva LP, MALDI-TOF MS profiling approach: how much can we get from it. Front. Plant Sci 2015;6:184.
Clark CM, Costa MS, Sanchez LM, Murphy BT. Coupling MALDI-TOF mass spectrometry protein and specialized metabolite analyses to rapidly discriminate bacterial function. Proc Natl Acad Sci U.S.A. 2018;115(19):4981-6.
Xun Y, Feng X, Xue Y, Feng L, et al. Profile of twenty- three human milk oligosaccharides in Han Chinese mothers throughout postpartum 1 year. J Food Quality 2022:9.
Zenhom M, Hyder A, de Vrese M, Heller KJ, Roeder T, Schrezenmeir J. Prebiotic oligosaccharides reduce proinflammatory cytokines in intestinal Caco-2 cells via activation of PPARgamma and peptidoglycan recognition protein 3. J Nutr 2011;141(5):971-7.
Huang Z, Li Y, Luo Y, Guo H. Human milk oligosaccharides 3’-sialyllactose and 6’-sialyllactose protect intetine against necrotizing enterocolitis damage induced by hypoxia. J Func Food 2021;86:104708.
Lianghui Cheng MBGK, AndreGroeneveld, ArjenNauta, Paulde Vos. Human milk oligosaccharides and its acid hydrolysate LNT2 show immune modulatory effects via TLRs in a dose and structure-dependent way. J Func Foods 2019;59:174-84.
Holscher HD, Bode L, Tappenden KA. Human Milk Oligosaccharides Influence Intestinal Epithelial Cell Maturation In Vitro. J Pediatr Gastroenterol Nutr 2017;64(4):296-301.
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