Intestinal epithelial barrier integrity is crucial for maintaining gut homeostasis and preventing luminal inflammation. Disruption of tight junctions (TJs) by pro-inflammatory cytokines such as tumor necrosis factor-α and interferon-γ contributes to barrier dysfunction, a hallmark of disorders like inflammatory bowel disease. In this study, we investigated the protective effects of hydroxypropyl methylcellulose (HPMC), a widely used excipient and dietary fiber, against cytokine-induced epithelial barrier dysfunction in Caco-2 cell monolayers. HPMC treatment preserved transepithelial electrical resistance, reduced paracellular leakage of FITC-dextran, and significantly suppressed the secretion of inflammatory mediators, including interleukin (IL)-8, IL-1β, IL-6, and monocyte chemoattractant protein (MCP)-1. Furthermore, HPMC restored the expression and localization of TJ proteins zonula occludens-1 and occludin and attenuated NF-κB activation by inhibiting IκBα phosphorylation and subsequent nuclear translocation. These findings indicate that HPMC counteracts cytokine-driven epithelial barrier disruption through coordinated anti-inflammatory and barrier-stabilizing actions. Collectively, our results demonstrate that noncytotoxic HPMC (12.5–100 μg/mL) has potential as a safe pharmaceutical excipient and functional dietary fiber capable of supporting intestinal health under inflammatory conditions.
Al-SadiR, KhatibK, GuoS, et al.Occludin regulates macromolecule flux across the intestinal epithelial tight junction barrier. Am J Physiol Gastrointest Liver Physiol2011;300(6):G1054–G64; doi: 10.1152/ajpgi.00055.2011
2.
FanningAS, MaTY, AndersonJM. Isolation and functional characterization of the actin binding region in the tight junction protein ZO-1. FASEB J2002;16(13):1835–1837; doi: 10.1096/fj.02-0121fje
3.
RaoR. Occludin phosphorylation in regulation of epithelial tight junctions. Ann N Y Acad Sci2009;1165:62–68; doi: 10.1111/j.1749-6632.2009.04054.x
D’IncaR, Di LeoV, CorraoG, et al.Intestinal permeability test as a predictor of clinical course in Crohn’s disease. Am J Gastroenterol1999;94(10):2956–2960; doi: 10.1111/j.1572-0241.1999.01444.x
6.
BruewerM, LuegeringA, KucharzikT, et al.Proinflammatory cytokines disrupt epithelial barrier function by apoptosis-independent mechanisms. J Immunol2003;171(11):6164–6172; doi: 10.4049/jimmunol.171.11.6164
7.
GuoG, ShiF, ZhuJ, et al.Piperine, a functional food alkaloid, exhibits inhibitory potential against TNBS-induced colitis via the inhibition of IkappaB-alpha/NF-kappaB and induces tight junction protein (claudin-1, occludin, and ZO-1) signaling pathway in experimental mice. Hum Exp Toxicol2020;39(4):477–491; doi: 10.1177/0960327119892042
8.
PoritzLS, GarverKI, GreenC, et al.Loss of the tight junction protein ZO-1 in dextran sulfate sodium induced colitis. J Surg Res2007;140(1):12–19; doi: 10.1016/j.jss.2006.07.050
9.
BurdockGA. Safety assessment of hydroxypropyl methylcellulose as a food ingredient. Food Chem Toxicol2007;45(12):2341–2351; doi: 10.1016/j.fct.2007.07.011
10.
AlmoazenH, FeltonLR. Essentials of pharmaceutics. Pharmaceutical press; 2013, 772pp, $69.00 (softcover), ISBN 9780857111050. Am J Pharm Educ2013;77.
11.
SiepmannJ, PeppasNA. Modeling of drug release from delivery systems based on hydroxypropyl methylcellulose (HPMC). Adv Drug Deliv Rev2012;64:163–174; doi: 10.1016/j.addr.2012.09.028
12.
JyotiMA, ThaiVV, MinYK, et al.In vitro bioactivity and biocompatibility of calcium phosphate cements using Hydroxy-Propyl-Methyl-Cellulose (HPMC). Appl Surf Sci2010;257(5):1533–1539; doi: 10.1016/j.apsusc.2010.08.091
13.
MoghimiR, AliahmadiA, RafatiH. Antibacterial hydroxypropyl methyl cellulose edible films containing nanoemulsions of Thymus daenensis essential oil for food packaging. Carbohydr Polym2017;175:241–248; doi: 10.1016/j.carbpol.2017.07.086
14.
PolamapllyP, ChengYL, ShiXL, et al.3D printing and characterization of hydroxypropyl methylcellulose and methylcellulose for biodegradable support structures. Polymer (Guildf)2019;173:119–126; doi: 10.1016/j.polymer.2019.04.013
15.
ZhangJ, YangW, VoAQ, et al.Hydroxypropyl methylcellulose-based controlled release dosage by melt extrusion and 3D printing: Structure and drug release correlation. Carbohydr Polym2017;177:49–57; doi: 10.1016/j.carbpol.2017.08.058
16.
LiF, DuP, YangW, et al.Polysaccharide from the seeds of Plantago asiatica L. alleviates nonylphenol induced intestinal barrier injury by regulating tight junctions in human Caco-2 cell line. Int J Biol Macromol2020;164:2134–2140; doi: 10.1016/j.ijbiomac.2020.07.259
17.
LiY, TianX, LiS, et al.Total polysaccharides of adlay bran (Coix lachryma-jobi L.) improve TNF-alpha induced epithelial barrier dysfunction in Caco-2 cells via inhibition of the inflammatory response. Food Funct2019;10(5):2906–2913; doi: 10.1039/c9fo00590k
18.
TuJ, XuY, XuJ, et al.Chitosan nanoparticles reduce LPS-induced inflammatory reaction via inhibition of NF-kappaB pathway in Caco-2 cells. Int J Biol Macromol2016;86:848–856; doi: 10.1016/j.ijbiomac.2016.02.015
19.
KimH, BartleyGE, YoungSA, et al.Altered hepatic gene expression profiles associated with improved fatty liver, insulin resistance, and intestinal permeability after Hydroxypropyl Methylcellulose (HPMC) supplementation in diet-induced obese mice. J Agric Food Chem2013;61(26):6404–6411; doi: 10.1021/jf400545w
20.
KimH, BartleyGE, YoungSA, et al.HPMC supplementation reduces abdominal fat content, intestinal permeability, inflammation, and insulin resistance in diet-induced obese mice. Mol Nutr Food Res2012;56(9):1464–1476; doi: 10.1002/mnfr.201200082
21.
BaiY, HuangF, ZhangR, et al.Longan pulp polysaccharides relieve intestinal injury in vivo and in vitro by promoting tight junction expression. Carbohydr Polym2020;229:115475; doi: 10.1016/j.carbpol.2019.115475
22.
BaiY, JiaX, HuangF, et al.Structural elucidation, anti-inflammatory activity and intestinal barrier protection of longan pulp polysaccharide LPIIa. Carbohydr Polym2020;246:116532; doi: 10.1016/j.carbpol.2020.116532
23.
WangJ, ZhangC, GuoC, et al.Chitosan Ameliorates DSS-Induced ulcerative colitis mice by enhancing intestinal barrier function and improving microflora. Int J Mol Sci2019;20(22):5751; doi: 10.3390/ijms20225751
24.
LiangJ, LiH, ChenJ, et al.Dendrobium officinale polysaccharides alleviate colon tumorigenesis via restoring intestinal barrier function and enhancing anti-tumor immune response. Pharmacol Res2019;148:104417; doi: 10.1016/j.phrs.2019.104417
25.
Kandhaya-PillaiR, YangX, TchkoniaT, et al.TNF-alpha/IFN-gamma synergy amplifies senescence-associated inflammation and SARS-CoV-2 receptor expression via hyper-activated JAK/STAT1. Aging Cell2022;21(6):e13646; doi: 10.1111/acel.13646
26.
CaoM, WangP, SunC, et al.Amelioration of IFN-gamma and TNF-alpha-induced intestinal epithelial barrier dysfunction by berberine via suppression of MLCK-MLC phosphorylation signaling pathway. PLoS One2013;8(5):e61944; doi: 10.1371/journal.pone.0061944
27.
OhJH, KimSH, KwonOK, et al.Purpurin suppresses atopic dermatitis via TNF-α/IFN-γ-induced inflammation in HaCaT cells. Int J Immunopathol Pharmacol2022;36:3946320221111135; doi: 10.1177/03946320221111135
28.
YuM, WangQ, MaY, et al.Aryl hydrocarbon receptor activation modulates intestinal epithelial barrier function by maintaining tight junction integrity. Int J Biol Sci2018;14(1):69–77; doi: 10.7150/ijbs.22259
29.
LeeJH, KimS, KangM-C, et al.Intestinal epithelial barrier protective effect of nut oils (walnut, almond, pistachio, and pine nut) on TNF-α/IFN-γ induced damage in Caco-2 cell monolayers. J Funct Foods2023;111:105887; doi: 10.1016/j.jff.2023.105887
30.
Al-SadiR, BoivinM, MaT. Mechanism of cytokine modulation of epithelial tight junction barrier. Front Biosci (Landmark Ed)2009;14(7):2765–2778; doi: 10.2741/3413
31.
BruewerM, UtechM, IvanovAI, et al.Interferon-gamma induces internalization of epithelial tight junction proteins via a macropinocytosis-like process. Faseb J2005;19(8):923–933; doi: 10.1096/fj.04-3260com
32.
ChondrogiannisG, KastamoulasM, KanavarosP, et al.Cytokine effects on cell viability and death of prostate carcinoma cells. Biomed Res Int2014;2014:536049; doi: 10.1155/2014/536049
33.
LeeSH. Intestinal permeability regulation by tight junction: Implication on inflammatory bowel diseases. Intest Res2015;13(1):11–18; doi: 10.5217/ir.2015.13.1.11
34.
MaTY, IwamotoGK, HoaNT, et al.TNF-α-induced increase in intestinal epithelial tight junction permeability requires NF-κB activation. Am J Physiol Gastrointest Liver Physiol2004;286(3):G367–G376; doi: 10.1152/ajpgi.00173.2003
35.
MadaraJL, StaffordJ. Interferon-gamma directly affects barrier function of cultured intestinal epithelial monolayers. J Clin Invest1989;83(2):724–727; doi: 10.1172/JCI113938
36.
MitselouA, GrammeniatisV, VarouktsiA, et al.Proinflammatory cytokines in irritable bowel syndrome: A comparison with inflammatory bowel disease. Intest Res2020;18(1):115–120; doi: 10.5217/ir.2019.00125
37.
PapadakisKA, TarganSR. Role of cytokines in the pathogenesis of inflammatory bowel disease. Annu Rev Med2000;51:289–298; doi: 10.1146/annurev.med.51.1.289
38.
SchmitzH, FrommM, BentzelCJ, et al.Tumor necrosis factor-alpha (TNFalpha) regulates the epithelial barrier in the human intestinal cell line HT-29/B6. J Cell Sci1999;112 (Pt 1):137–146; doi: 10.1242/jcs.112.1.137
39.
SchneebergerEE, LynchRD. Structure, function, and regulation of cellular tight junctions. Am J Physiol1992;262(6 Pt 1):L647–L61; doi: 10.1152/ajplung.1992.262.6.L647
40.
SchulzkeJD, PloegerS, AmashehM, et al.Epithelial tight junctions in intestinal inflammation. Ann N Y Acad Sci2009;1165:294–300; doi: 10.1111/j.1749-6632.2009.04062.x
41.
StruyfS, GouwyM, DillenC, et al.Chemokines synergize in the recruitment of circulating neutrophils into inflamed tissue. Eur J Immunol2005;35(5):1583–1591; doi: 10.1002/eji.200425753
42.
WangF, GrahamWV, WangY, et al.Interferon-gamma and tumor necrosis factor-alpha synergize to induce intestinal epithelial barrier dysfunction by up-regulating myosin light chain kinase expression. Am J Pathol2005;166(2):409–419; doi: 10.1016/s0002-9440(10)62264-x
43.
WangR, LiZ, LiuS, et al.Global, regional and national burden of inflammatory bowel disease in 204 countries and territories from 1990 to 2019: A systematic analysis based on the Global Burden of Disease Study 2019. BMJ Open2023;13(3):e065186; doi: 10.1136/bmjopen-2022-065186
